Artykuły w czasopismach: „Antigen antibody reaction”

1

Montero, Claudio. "The Antigen-Antibody Reaction in Immunohistochemistry". Journal of Histochemistry & Cytochemistry 51, nr 1 (styczeń 2003): 1–4. http://dx.doi.org/10.1177/002215540305100101.

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The problems of major concern in immunohistochemical practice are discussed in the following order: (a) the mechanism of the Ag-Ab reaction in fixed tissue as opposed to the in vitro reaction; (b) the chemistry of fixation and its influence on the final result of the immunohistochemical reaction; (c) the various procedures used for antigen retrieval in formaldehyde-fixed tissue; and (d) the consideration of the possible mechanism underlying heat-induced antigen retrieval. Suggestions for further work to attempt a clarification of the mechanism involved in the Ag-Ab reaction in immunohistochemistry resorting to existing histochemical methods for the demonstration of protein side groups are presented, together with some examples already published.

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Kobayashi, T., I. Yokoyama, H. Ogawa, T. Nagasaka, D. Liu, T. Kato, T. Tokoro i in. "Suppression of antigen-antibody reaction in xenotransplantation". Transplantation Proceedings 32, nr 2 (marzec 2000): 287–88. http://dx.doi.org/10.1016/s0041-1345(99)00959-8.

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Chernykh, Oleg Yu, Elmira A. Yanikova, Mikail M. Mikailov, Akhmed A. Khalikov i Atiya T. Gulieva. "Indirect hemagglutination reaction in ram infectious epididymitis for indication of Brucella ovis antigen in biomaterial". Veterinaria Kubani, nr 5 (30.10.2020): 23–25. http://dx.doi.org/10.33861/2071-8020-2020-5-23-25.

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The reaction of indirect hemagglutination with an antibody erythrocyte diagnosticum is one of the promising methods for identifying the causative agent of infectious diseases or antigen. This preparation has not yet been developed for the diagnosis of infectious epididymitis of rams. The results of the preparation of an antibody erythrocyte diagnosticum in the reaction of indirect hemagglutination for the detection of the causative agent of infectious epididymitis in various biological material are presented in the article. As a result of scientific research, authors developed the method for obtaining an original Brucella ovis antibody diagnosticum for the reaction of indirect hemagglutination by sensitizing sheep erythrocytes with hyperimmune Brucella ovis serum using alizarin blue indicator of the epididymitic pathogen or antigen in biomaterial as the conjugate. Studies have shown the specificity and higher sensitivity of the indirect hemagglutination reaction with the antibody erythrocyte diagnosticum, compared with the antibody neutralization reaction and the bacteriological method, and its suitability for the indication of Brucella ovis antigen in biomaterial and environmental objects. It was also found that the reaction of indirect hemagglutination with antibody diagnosticum is much faster than the reaction of neutralization of antibodies and the bacteriological method. Research to improve the diagnosis of infectious ram epididymitis caused by Brucella ovis has been completed with the development of the antibody erythrocyte diagnosticum for the indirect hemagglutination reaction.The high specificity and activity of this preparationwas established by the authors. As a result of the studies carried out to test the diagnostic value of the antibody diagnosticum, a higher sensitivity of the indirect hemagglutination reaction using the new antibody diagnosticum, compared with the neutralization reaction of antibodies and the bacteriological method, and the suitability of using the diagnosticum for the indication of the Brucella ovis antigen in biomaterial, was established, which meets the requirements for express methods for detecting antigens in pathological material.

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Grushevskaja, G. V., i A. I. Khmelnitsky. "Sequence of bifurcations in an ‘antigen-antibody’ reaction". Journal of Molecular Recognition 9, nr 5-6 (październik 1996): 570–74. http://dx.doi.org/10.1002/(sici)1099-1352(199634/12)9:5/6<570::aid-jmr304>3.0.co;2-k.

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MALMBORG, A. C., A. MICHAELSSON, M. OHLIN, B. JANSSON i C. A. K. BORREBAECK. "Real Time Analysis of Antibody-Antigen Reaction Kinetics". Scandinavian Journal of Immunology 35, nr 6 (czerwiec 1992): 643–50. http://dx.doi.org/10.1111/j.1365-3083.1992.tb02970.x.

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Allan, J. C., i P. S. Craig. "Partial characterization and time course analysis of Hymenolepis diminuta coproantigens". Journal of Helminthology 68, nr 2 (czerwiec 1994): 97–103. http://dx.doi.org/10.1017/s0022149x00013596.

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AbstractAn analysis of Hymenolepis diminuta specific antigens in infected rat faeces was carried out. Using a capture type antibody sandwich ELISA assay based on a hyperimmune rabbit anti-worm somatic antisera it was demonstrated that, although antigen was present in faeces before patency, the onset of egg production led to a sharp increase in the levels of parasite antigen in the faeces. Levels of antigen in host faeces were independent of worm burden. Parasite eggs did not contribute significantly to faecal antigen levels. Western blot analysis indicated a number of highly specific antigens at around Mr 69,000, Mr 37,000, Mr 50,000 and Mr 27,000 with a low molecular weight smear at between Mr 30,000 and Mr 34,000 present in the faeces of H. diminuta infected rats. Some cross reaction occurred with an antigen of around Mr 66,000 in the faeces of non H. diminuta infected rodents. Antibody activity against this antigen was removed by affinity adsorption of the antibody solution against normal rat faeces.

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Wajer, S. D., i H. K. Charles. "A SEM analysis of thin indium films for immunoassay applications". Proceedings, annual meeting, Electron Microscopy Society of America 45 (sierpień 1987): 938–39. http://dx.doi.org/10.1017/s0424820100128973.

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Indium metal films have been used as host substrate materials for biological immunoassays. Indium film immunoassays involve the optical detection of specific antibody-antigen reactions using either transmitted or reflected white light. In the immunoassay process, drops of a purified antibody solution are placed on 5 to 25 nm thick indium films supported on a glass substrate (i.e., indium slide). After incubation, the antibody solution is washed off, leaving visible spots of reactive antibody. The slide is then coated with an unrelated protein which covers any non-antibody areas thus making the whole slide appear optically uniform. Placing the indium slide in an appropriate antigen solution will cause a visible response, since the antibody-antigen reaction complex will contain two layers of protein and will be darker than the surrounding background. In addition to the biological reaction sensitivities, the indium film immunoassay response is critically dependent on the bulk and surface properties of the indium film.

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8

Hohsaka, Takahiro, Kazuo Kawashima i Masahiko Sisido. "Photoswitching of NAD+-mediated enzyme reaction through photoreversible antigen-antibody reaction". Journal of the American Chemical Society 116, nr 1 (styczeń 1994): 413–14. http://dx.doi.org/10.1021/ja00080a064.

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YANAGI, AKIRA, i HIROAKI YAMAMOTO. "Monoclonal antibody against a conjugation-specific nuclear antigen in Paramecium caudatum". Journal of Cell Science 95, nr 2 (1.02.1990): 287–91. http://dx.doi.org/10.1242/jcs.95.2.287.

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To understand molecular mechanisms controlling the sexual reproduction process (conjugation) in Paramecium caudatum, we have tried to detect conjugation-specific antigens with monoclonal antibodies. We obtained a monoclonal antibody (CSN-1) against an antigen that appears in the nuclei only during conjugation. This nuclear antigen began to appear both in micro- and macronuclei at micronuclear stage II or III early in the conjugation process (4 h after the mating reaction at 25°C). In the macronucleus, the nuclear antigen persisted until the stage of macronuclear fragmentation (about 35 h) and then disappeared before degeneration of the macronuclear fragments. In the micronucleus, the antigen existed until the crescent stage (stage V) of the first meiotic division (8h). The antigen in the micronucleus disappeared after the crescent stage but reappeared again in the eight nuclei produced by the third postzygotic division (25 h). Then it persisted only in the four macronuclear anlagen differentiated from the eight nuclei (about 30 h). When exconjugant cells had undergone two successive postconjugational cell divisions and thus possessed only one new macronucleus as in the vegetative cells, the antigen disappeared completely from the new macronucleus in most cells. These cells without the antigen began to appear about 50 h after the mating reaction. As the antigen is specific to conjugation and localized in nuclei, it may play some important role(s) in the conjugation process.

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Sudjana, Dina, Lia Amalia i Saepul Adnan. "Preliminary Immunochemical Studies to Detect Lard". Indonesian Journal of Halal Research 2, nr 1 (1.03.2020): 8–12. http://dx.doi.org/10.15575/ijhar.v2i1.7819.

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Detection of lard in food using immunochemical methods has been carried out. This method has been applied and developed in food analysis. The purpose of this study was to detect the presence of lard in food. The method used was immunochemical which in this case is tested to its application in food analysis. The principle method is based on the antigen-antibody reaction, between lipids as antigens that can be deposited by antibodies in the agar medium. Antibodies were obtained by inducing lard into the blood vessels of rabbits. This method is based on the antigen-antibody reaction between fat as an antigen and antibodies that contain anti-lard as reagents and the occurrence of precipitation in agar media. The antibody was obtained by inducing lard, pork broth, and pig plasma into the blood vessels of rabbits. The results of this study were significant.

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Ye, Zhixiang, i Zongqing Huang. "Study of antigen and antibody reaction in electrochemical reactions with ellipsometric spectroscopy". Chinese Science Bulletin 42, nr 2 (styczeń 1997): 176. http://dx.doi.org/10.1007/bf03182799.

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Liang, Jiahui, Xiaotian Ye, Jiang He, Jing Liu, Yaoxiong Huang i Feiyue Xing. "Novel Immune Microlens Imaging for Detection of Antigen and Antibody". Journal of Immunology Research 2019 (18.04.2019): 1–8. http://dx.doi.org/10.1155/2019/5474519.

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Detection and analysis of antigen-antibody reaction is one of the most critical detection techniques in the fields of medicine, biology, environmental science, and food safety. Traditional and classical methods for detecting antigen and antibody encounter many problems, such as time-consuming, high cost, and low accuracy. A novel immune microsphere imaging technique by the microlens is used to test the changes of refractive index before and after antigen-antibody reaction. It can quickly perform qualitative and quantitative determination for antigen-antibody reaction without any labeling, premodification, postwashing, and expensive enzymes. Here, we feature and discuss its principle and advantages, structure of a microlens immunoassay instrument, and potential in measuring clinical samples. It is promising to be developed for application to diagnosis of clinical diseases.

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13

Sakata, H., H. Morioka, M. kinjo i M. Tamura. "1P089 Antigen-Antibody-Reaction Analysis by Fluorescence Correlation Spectroscopy". Seibutsu Butsuri 44, supplement (2004): S52. http://dx.doi.org/10.2142/biophys.44.s52_1.

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Dybkaer, René. "Scales for Measurement Based on an Antigen-Antibody Reaction". Scandinavian Journal of Clinical and Laboratory Investigation 51, sup205 (styczeń 1991): 55–62. http://dx.doi.org/10.3109/00365519109104602.

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Yoshimi, Yasuo, Hirotaka Haramoto, Takehiro Miyasaka i Kiyotaka Sakai. "Cathodic Electrochemiluminescence of Luminol Enhanced by Antibody-Antigen Reaction." JOURNAL OF CHEMICAL ENGINEERING OF JAPAN 29, nr 5 (1996): 851–57. http://dx.doi.org/10.1252/jcej.29.851.

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Skoug, J. W., i H. L. Pardue. "Effects of reaction variables on nephelometric and turbidimetric responses for the immunochemical reaction of immunoglobulin G." Clinical Chemistry 34, nr 2 (1.02.1988): 300–308. http://dx.doi.org/10.1093/clinchem/34.2.294.

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Abstract Here we report a kinetic study of the immunoprecipitin reaction involving immunoglobulins G, A, and M. We used stopped-flow mixing adapted for simultaneous monitoring of nephelometric and turbidimetric signals for some studies and centrifugal mixing for others. The variables having the most significant effects on the kinetic responses in the regions of excess antibody and excess antigen are polyethylene glycol concentration, ionic strength, and the ratio of antigen to antibody. We document effects of these variables on maximum velocities and signal changes over a fixed-time interval for both monitoring modes and on the maximum signal change for nephelometry. We use response-surface methodology to help identify interactive effects among these variables (polyethylene glycol, NaCl, and antibody concentrations) and to select the best combination to use to quantify antigen in all regions of the immunoprecipitin curve. We also observe that turbidimetric responses are more reproducible and much simpler than are nephelometric responses. Implications of these results for quantification of the immunoglobulins are discussed.

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Fukuizumi, Takaki, Hiromasa Inoue, Toshiyuki Tsujisawa i Choji Uchiyama. "Streptococcus sobrinus Antigens That React to Salivary Antibodies Induced by Tonsillar Application of Formalin-Killed S. sobrinus in Rabbits". Infection and Immunity 68, nr 2 (1.02.2000): 725–31. http://dx.doi.org/10.1128/iai.68.2.725-731.2000.

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ABSTRACT We previously found that tonsillar application of antigen induces a strong antibody response to Streptococcus sobrinus in saliva and blood plasma. Rabbits immunized against S. sobrinus by tonsillar application were highly resistant to experimental dental caries triggered by oral inoculation of livingS. sobrinus organisms with sucrose. In the present study, we examined the reaction of S. sobrinus antigens to the antibodies induced by the tonsillar application of S. sobrinus AHT-k in rabbits and compared them to those antibodies induced by intramuscular injection. In an enzyme-linked immunosorbent assay using ultrasonic fragments from mutans group streptococci, the saliva and blood plasma selectively reacted to S. sobrinusAHT-k (serotype g) and serologically related streptococci (serotypes a, d, and h) in the sixth week after tonsillar application, whereas the blood plasma in the sixth week after intramuscular injection reacted to the unrelated streptococci (serotypes b, c, e, and f) in addition to the aforementioned streptococci. The antibody reactivity induced after tonsillar application was not lost after treatment of the antigen with heat or proteinase digestion, whereas these treatments resulted in a 70% decrease of the antibody reactivity induced by intramuscular injection. The inhibition by haptenic sugars and the decrease in immunoreactivity by heat treatment and proteinase digestion suggested that 80% of the antibodies induced by tonsillar application reacted to saccharides. These saccharide antigens appeared to be involved in a specific reaction with S. sobrinus-specific streptococci and a selective reaction with serologically related streptococci. These antigens are probably involved in anticaries reactions in experimental dental caries.

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Vancutsem, E., F. Echahidi, K. Van Geel, G. Muyldermans, O. Soetens i A. Naessens. "Production of Recombinant Antigens of Ureaplasma parvum Serotypes 3 and 6 for Development of a Serological Assay". Clinical and Vaccine Immunology 15, nr 3 (19.12.2007): 447–51. http://dx.doi.org/10.1128/cvi.00379-07.

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ABSTRACT Recombinant antigens of Ureaplasma parvum serotypes 3 and 6 were produced in order to develop a serological assay for Ureaplasma antibody detection. The genes of the multiple banded antigen (MBA) were amplified by PCR and cloned in a pTrcHis TOPO plasmid. Purified recombinant proteins were evaluated in Western blotting and enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies and human sera. Our approach was successful in the production of the recombinant MBAs (rMBAs) for serotypes 3 and 6. The antigens tested positive with serotype-specific monoclonal antibodies in Western blotting and in ELISA. Prominent reactions were detected with the rMBAs and their homologous monoclonal antibodies. Strong cross-reactions were visible in ELISA between rMBA 3 and the monoclonal antibodies from the other U. parvum serotypes. A weak cross-reaction was seen with rMBA 3 and the monoclonal antibody from serotype 4. rMBA 6 showed cross-reaction only with the monoclonal antibody from U. parvum serotype 1. Fifty-one percent of the sera obtained from culture-positive women reacted with one or both rMBAs. Only three (15%) of the sera from culture-negative women reacted with the rMBA. The positive reactions were observed only with rMBA 6. These preliminary tests showed the potential usefulness of the rMBAs produced for detecting an antibody response against Ureaplasma antigens.

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Khosravi, Afra, Sobhan Ghafourian, Morteza Shamsi, Nourkhoda Sadeghifard, Abbas Maleki i Ebrahim Babaahmadi. "Cross-Reaction between the Crude Hydatid Cyst Fluid Antigens of Human and Animals Origin in Response to Human IgG Class and Subclasses". Journal of Parasitology Research 2012 (2012): 1–8. http://dx.doi.org/10.1155/2012/947948.

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The current work aimed to evaluate the cross-reactivity of human immune sera against crude hydatid fluid antigens of sheep, human, mouse, cattle, as well as B fraction of cystic fluid antigen. 30 balb/c mice were infected with sheep hydatid cyct fluid antigen containing protoscolex after the viability of these protoscolices was assessed. ANOVA was used to test the difference of themean of optical density (OD) values among case and control groups. The highest human IgG class antibody was against antigen B (0.93) and the lowest against cattle HCF antigen (0.32). The differences between responses to these antigens were statistically significant (P<0.001). The sensitivity and specificity of ELISA test used for evaluating the responses of human total IgG to different hydatid cyst fluid (HCF) antigens among the case and control groups were 100 and 95.8%, respectively. Cross-reaction of human IgG class and subclasses responses was found almost for all the antigens with the best reaction against human and cattle (HCF) antigens and antigen B using a ratio of mean OD value to each antigen divided by the cut-off point value for the same antigen. Human sera showed a considerable cross-reactivity against all antigens by using ELISA.

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Sasidharan, S., i A. M. Uyub. "Antibody Response toHelicobacter pyloriExcretory Antigen and the Cross Reaction Study". Journal of Immunoassay and Immunochemistry 30, nr 1 (31.12.2008): 70–81. http://dx.doi.org/10.1080/15321810802569477.

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Hidayat, Rachmat, i Patricia Wulandari. "Enzyme Linked Immunosorbent Assay (ELISA) Technique Guideline". Bioscientia Medicina : Journal of Biomedicine and Translational Research 5, nr 2 (29.01.2021): 352–58. http://dx.doi.org/10.32539/bsm.v5i2.228.

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A B S T R A C TELISA (Enzyme-linked immunosorbent assay) is a technique used to assessthe quantification of peptide, protein, antibody and hormone levels, basedon the principle of antigen-antibody binding. In the ELISA technique, antigenimmobilization will be carried out on a solid surface, then bound withantibodies to form an antigen-antibody bond complex, where the antigen-antibody complex is bound to the enzyme. The detection signal in the formof a color change will be formed due to the reaction between the enzyme andthe substrate.

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Britton, W. J., L. Hellqvist, A. Basten i A. S. Inglis. "Immunoreactivity of a 70 kD protein purified from Mycobacterium bovis Bacillus Calmette-Guerin by monoclonal antibody affinity chromatography." Journal of Experimental Medicine 164, nr 3 (1.09.1986): 695–708. http://dx.doi.org/10.1084/jem.164.3.695.

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The protein antigens from Mycobacterium bovis (BCG), M. tuberculosis, and M. leprae share a number of common determinants. We have used a murine mAb (L7) recognizing such a determinant on a protein of Mr 70,000 to purify this antigen from M. bovis sonicate by affinity chromatography. Enrichment of the protein in column eluates was confirmed by immunoblotting and in antigen inhibition assays. After radiolabelling with 125I, the protein could be immunoprecipitated with human lepromatous leprosy sera. Stimulation of peripheral blood mononuclear cells from BCG-vaccinated and naturally mantoux-positive individuals induced proliferation and IFN-gamma secretion, while intradermal injection of purified antigen into the same subjects resulted in a delayed-type hypersensitivity reaction. Thus, the 70,000 molecule carried epitopes capable of reacting with B cells, and eliciting a potentially protective T cell response. The first 15 N-terminal residues were sequenced using a gas-phase sequenator.

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Setiawan, Made, Agus Sjahrurachman, Fera Lbrahim i Agus Suwandono. "Differences of antigenic profiles on immunoblotting of wild type measles virus and vaccine virus in Indonesia". Paediatrica Indonesiana 48, nr 6 (15.09.2016): 364. http://dx.doi.org/10.14238/pi48.6.2008.364-73.

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Background Measles virus has a negative, single strand RNAgenome which codes for six important structural proteins. Thegenes of the wild type measles virus have many variances hencethe nucleotide sequences of each wild type virus and vaccine virusare different. This differences lead to the antigenic differencesbetween wild type and vaccine virus.Objective The purpose of this research is to investigate thedifferences in the antigenic profiles on immunoblotting betweenwild type and vaccine virus.Results The analysis results are 1) the antigen ofCAM-70 vaccinevirus was less able in cross reacting with the antibodies from G2,G3, 09, CAM-70 and Schwarz; 2) The antibody aga inst CAM-70 was only able to cross react with antigens of N protein and afew of antigens ofF proteins; 3) The wild type virus were veryimmunogenic, hence the antibody titers were very high; 4) TheCAM-70 and MMR vaccine virus were less immunogenic, hencetheir antibody were very low; 5) The antibody responses thatalways occurred from all immunized mice serum were antibodyfor N and F proteins. However, the antibody against CAM-70vaccine virus was still able to react with wild type virus (G2, G3and 09).Conclusion All antigen-antibody reaction on immunoblottingresulted in different profiles especially between wild type virusand CAM-70 vaccine virus. Although CAM-70 vaccine virusshowed clear differences compared to G2, G3 and 09 genotypes,antibodies against CAM-70 were still able to cross react withantigens from other genotypes (G2, G3 and D9).

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Vilches-Moure, José G., i José A. Ramos-Vara. "Comparison of Rabbit Monoclonal and Mouse Monoclonal Antibodies in Immunohistochemistry in Canine Tissues". Journal of Veterinary Diagnostic Investigation 17, nr 4 (lipiec 2005): 346–50. http://dx.doi.org/10.1177/104063870501700407.

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Rabbit monoclonal (RM) antibodies appear to have higher affinity for antigens than mouse monoclonal (MM) antibodies. However, RM antibodies have not been used in veterinary diagnostic immunohistochemistry. The authors compared reactivities of RM and MM antibodies on formalin-fixed, paraffin-embedded canine tissues, targeting 11 different antigens: CD3, CD79a, calcitonin, calretinin, chromogranin A, COX-2, estrogen receptor, Ki67, progesterone receptor, synaptophysin, and vimentin. Paraffin-embedded tissue sections were processed by 1 of 2 antigen-retrieval methods: 1) proteinase K digestion or 2) steam heat in citrate buffer. An additional set of slides did not receive antigen retrieval. Immunostaining was performed using an automated stainer, and scores were assigned to the different dilutions and antigen-retrieval methods on the basis of staining intensity and number of positive cells. Steam heat was usually the best antigen-retrieval method. The optimal dilution for each antibody was that which resulted in the highest specific staining and the lowest nonspecific (background) staining. The RM or MM antibodies yielded a specific reaction for all antigens examined except calretinin. The RM and MM antibodies yielded a specific reaction for 4 antigens only: COX-2, Ki67, synaptophysin, and vimentin. Three antigens (CD3, chromogranin A, and progesterone receptor) were detected only with RM antibodies, whereas the other 3 (CD79a, calcitonin, estrogen receptor) were detected only with MM antibodies. The results of this study differed from those reported for human tissues by the manufacturers of the antibodies. These results emphasize that, regardless of manufacturers' recommendations, each antibody must be individually standardized and validated before routine use in canine tissues.

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Wang, Yanan, Hanna Fotina i Alexsey Fotin. "Design of antigen synthesis and identification of its artificial antigen for zearalenone". Ukrainian Journal of Veterinary and Agricultural Sciences 4, nr 2 (21.06.2021): 7–12. http://dx.doi.org/10.32718/ujvas4-2.02.

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Background: study aimed to modify the ZEN molecule and conjugate the carrier protein to prepare a complete antigen. To lay the foundation for the preparation of ZEN monoclonal antibodies. Methods: The carbonyl group at the 7 position of ZEN molecule was modified by deuteration reaction. The immunogen and antigen were synthesized by EDC method and mixed acid anhydride method, and the complete antigen was identified by UV, IR and electrophoresis. Antisera were obtained after immunization of animals, and an antiserum was designed by ELISA. Results: The immunogens were identified by UV, IR and electrophoresis, ZEN-BSA was successfully synthesized. The ratio of ZEN-BAS to ZEN was calculated to be 1 : 13. When the antibody serum was detected, the titer of the antibody reached 1:(6.4×103). Conclusion: This study demonstrated that the OAE method is preferable in preparing the ZEN. These findings lay the material and technical foundation for the preparation of anti-ZEN monoclonal antibody

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Parlane, Natalie A., Katrin Grage, Jason W. Lee, Bryce M. Buddle, Michel Denis i Bernd H. A. Rehm. "Production of a Particulate Hepatitis C Vaccine Candidate by an Engineered Lactococcus lactis Strain". Applied and Environmental Microbiology 77, nr 24 (7.10.2011): 8516–22. http://dx.doi.org/10.1128/aem.06420-11.

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ABSTRACTVaccine delivery systems based on display of antigens on bioengineered bacterial polyester inclusions can stimulate cellular immune responses. The food-grade Gram-positive bacteriumLactococcus lactiswas engineered to produce spherical polyhydroxybutyrate (PHB) inclusions which abundantly displayed the hepatitis C virus core (HCc) antigen. In mice, the immune response induced by this antigen delivery system was compared to that induced by vaccination with HCc antigen displayed on PHB beads produced inEscherichia coli, to PHB beads without antigen produced inL. lactisorE. coli, or directly to the recombinant HCc protein. Vaccination site lesions were minimal in all mice vaccinated with HCc PHB beads or recombinant protein, all mixed in the oil-in-water adjuvant Emulsigen, while vaccination with the recombinant protein in complete Freund's adjuvant produced a marked inflammatory reaction at the vaccination site. Vaccination with the PHB beads produced inL. lactisand displaying HCc antigen produced antigen-specific cellular immune responses with significant release of gamma interferon (IFN-γ) and interleukin-17A (IL-17A) from splenocyte cultures and no significant antigen-specific serum antibody, while the PHB beads displaying HCc but produced inE. colireleased IFN-γ and IL-17A as well as the proinflammatory cytokines tumor necrosis factor alpha (TNF-α) and IL-6 and low levels of IgG2c antibody. In contrast, recombinant HCc antigen in Emulsigen produced a diverse cytokine response and a strong IgG1 antibody response. Overall it was shown thatL. lactiscan be used to produce immunogenic PHB beads displaying viral antigens, making the beads suitable for vaccination against viral infections.

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Saccani Jotti, G., M. Fontanesi, A. Colombo, M. Valtolina i A. Tardini. "Localization of Mucinous - like Carcinoma Associated Antigen (MCA) in Breast Pathology: Comparison with Carcinoembryonic Antigen (CEA) and Tissue Polypeptide Antigen (TPA)". International Journal of Biological Markers 5, nr 3 (lipiec 1990): 145–52. http://dx.doi.org/10.1177/172460089000500308.

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The topographic distribution of a mucinous-like cancer antigen (MCA) recognized by a monoclonal antibody b-12 (MAb b-12) was assessed in benign (38) and malignant (66) breast tissues. The reactivity of MAb b-12 showed a good selectivity for breast tissues, reacting both with normal tissues and breast cancer. The degree of MCA expression was evaluated in the various groups of breast pathology adopting quantitative criteria of assessment. With the criteria of evaluation adopted, strong staining was observed in 71.4% breast carcinomas. The most positive reaction was demonstrated in mucinous carcinoma. MCA distribution in breast tissue was compared with the distribution of two other antigens, CEA and TPA. Reactivity of MAb b-12 was higher than the reactivity shown by the anti-CEA and anti-TPA antibodies.

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Fadeyi, Emmanuel A., Tawfeq Naal, Mary Green, Julie H. Simmons, Mary Rose Jones i Gregory J. Pomper. "Anti-M–Induced Delayed Hemolytic Transfusion Reaction". Laboratory Medicine 51, nr 4 (22.11.2019): 426–29. http://dx.doi.org/10.1093/labmed/lmz078.

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Abstract Background Anti-M is most often assumed to be naturally occurring and can be comprised of a mixture of predominantly immunoglobulin(Ig)M with a lesser IgG component. Anti-M-antibodies usually do not react at 37°C and therefore are considered to be of little clinical significance. Methods A 28-year-old man presented with hemorrhagic shock from numerous injuries sustained in a motor vehicle collision. The patient received several units of red blood cells (RBCs). The antibody screen, the direct antiglobulin test (DAT), and the RBC genotype were sent for laboratory evaluation. Results A total of 12 days after the first antibody screening result was negative (7 days after transfusion), the lowest hemoglobin value was 5.5 g per dL, and we observed a positive antibody screening result and DAT with immunoglobulin (Ig)G anti-M identified. After transfusion of 4 units of M antigen–negative RBC, the post-transfusion hemoglobin level increased to 8.9 g per dL. Conclusion Obtaining M antigen–negative compatible RBCs is necessary in patients demonstrating IgG anti-M in plasma.

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Wang, B. L., G. F. Springer i M. W. Kaufman. "Concurrent immunohistochemical staining of tumor-infiltrating lymphocytes and carcinoma-associated T (Thomsen-Friedenreich)/Tn antigens in human breast carcinoma." Journal of Histochemistry & Cytochemistry 44, nr 2 (luty 1996): 187–91. http://dx.doi.org/10.1177/44.2.8609376.

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The composition of tumor-infiltrating lymphocytes (TIL) often reflects the host's immune response to the tumor. To study the relationship of TIL and carcinoma-associated T/Tn antigens in breast carcinoma, a straightforward concurrent immunoenzyme staining procedure was developed. Fresh tissue was directly fixed in a zinc-based fixative to preserve lymphocyte markers and then routinely embedded in paraffin. The TIL subtypes in the sections were identified in the first immunostaining cycle by reaction with a monoclonal antibody (MAb) to lymphocyte markers CD3, CD4, CD8, CD19, or CD56, followed by a modified avidin-biotin procedure and diaminobenzidine tetrahydrochloride-H2O2 for color development. This was followed by paraformaldehyde fixation to block antibody crossreactivity. The T and Tn antigens on carcinoma cells were then demonstrated in a second staining cycle by reaction with an MAb against T or Tn antigen, followed by an indirect immunoalkaline phosphatase procedure and corresponding substrate systems for color development. The distinguishable brown color for TIL and blue or red color for T or Tn antigen enabled us to identify the TIL subsets and to describe their relations with T/Tn antigen expression in situ. This approach may contribute to better understanding of the patients' immune defenses against their tumor and aid in prognostication.

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Aryal, G. "Immunohistochemistry". Journal of Pathology of Nepal 5, nr 10 (14.09.2015): I. http://dx.doi.org/10.3126/jpn.v5i10.15660.

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Immunohistochemistry (IHC) or immunocytochemistry is a method of localizing specific antigen in tissue or cells based on antigen antibody reaction. IHC is the way of validating morphological findings. It helps in tumor diagnosis and classification, identify prognostic and predictive markers. IHC has a long history that dates back more than 70 years, when Coons1 first developed immunofluroscence technique to detect corresponding antigen in frozen tissue section. At oxford, Taylor and Burns2 developed the first successful demonstration of antigens in routinely processed formalin fixed paraffin-embedded sections. Since the early 1990s, IHC has been applied in routine formalin fixed paraffin embedded tissue. 3-4 Validation of reagents, protocols, controls and staining results are vital steps of IHC. The basic principles and protocols for fresh-frozen tissue sections are the same as those for paraffin sections, except that the antigen retrieval and dewaxing procedures are not required for frozen tissue sections. Titrations may also differ and must be separately optimized. Basic principles: Antigen-antibody recognition is based on the three-dimensional (3D) structure of protein or some other antigen, which may be compromised by formalin-induced modification of protein conformation (“masking”) but is restored in part by Antigen retrieval. Anti-A antibody binds specifically to antigen A in the tissue section. Antigen B (B) is depicted as a second antigenic determinant that is part of the anti-A molecule; anti-B antibody, made in a second species, will bind to this determinant. Thus anti-B, the so-called secondary antibody, can be used to locate the site of binding of anti-A, the primary antibody, in a tissue section. Basic IHC procedure Antigen retrieval (AR)-Enzymatic digestion (proteinase or trypsin), heat treatment (Microwave, water bath or autoclave) Blocking of non-specific background staining Incubation with primary antibody in humidity chamber Add avidin-biotin-peroxidase complex, which binds to secondary antibody Add 3, 3’ diaminobenzidine (DAB) as a chromagen (color changing reagent), with hematoxylin (Mayer's) counterstaining Theranostic Application IHC is becoming increasingly important for the evaluation of predictive markers that can help select patients who may respond to particular targeted therapies. Some of these CD117 for GI stromal tumors, Herceptin (Genentech, South San Francisco, CA) for HER2-positive breast cancers, and rituximab for CD20-positive lymphomas.

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Chitarra, V., P. M. Alzari, G. A. Bentley, T. N. Bhat, J. L. Eisele, A. Houdusse, J. Lescar, H. Souchon i R. J. Poljak. "Three-dimensional structure of a heteroclitic antigen-antibody cross-reaction complex." Proceedings of the National Academy of Sciences 90, nr 16 (15.08.1993): 7711–15. http://dx.doi.org/10.1073/pnas.90.16.7711.

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Toyokuni, Shinya, Waka Kawaguchi, Shinya Akatsuka, Makoto Hiroyasu i Hiroshi Hiai. "Intermittent microwave irradiation facilitates antigen-antibody reaction in Western blot analysis". Pathology International 53, nr 4 (kwiecień 2003): 259–61. http://dx.doi.org/10.1046/j.1320-5463.2003.01465.x.

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MCGILLIVERY, DJ, RE MORTON, WK YONG, GG RIFFKIN, C. JOHNSON i B. ADLER. "Confocal microscopy of the antigen-antibody reaction site in nematode larvae". Australian Veterinary Journal 69, nr 2 (luty 1992): 41–42. http://dx.doi.org/10.1111/j.1751-0813.1992.tb07438.x.

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Fukuyama, Masataka, Shinya Yamatogi, Hao Ding, Mizue Nishida, Chika Kawamoto, Yoshiteru Amemiya, Takeshi Ikeda i in. "Selective Detection of Antigen-Antibody Reaction Using Si Ring Optical Resonators". Japanese Journal of Applied Physics 49, nr 4 (20.04.2010): 04DL09. http://dx.doi.org/10.1143/jjap.49.04dl09.

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KAGAYA, Masami, i Yoichi AKAGAMI. "J164016 Rapid antigen-antibody reaction using electric field noncontact stirring technology". Proceedings of Mechanical Engineering Congress, Japan 2012 (2012): _J164016–1—_J164016–2. http://dx.doi.org/10.1299/jsmemecj.2012._j164016-1.

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Sakamoto, Satoshi, Kenshi Omagari, Yoshinori Kita, Yusuke Mochizuki, Yasuyuki Naito, Shintaro Kawata, Sachiko Matsuda i in. "Magnetically Promoted Rapid Immunoreactions Using Functionalized Fluorescent Magnetic Beads: A Proof of Principle". Clinical Chemistry 60, nr 4 (1.04.2014): 610–20. http://dx.doi.org/10.1373/clinchem.2013.211433.

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Abstract BACKGROUND Accurate detection and monitoring of disease-related biomarkers is important in understanding pathophysiology. We devised a rapid immunoreaction system that uses submicrometer polymer-coated fluorescent ferrite (FF) beads containing both ferrites (magnetic iron oxide) and fluorescent europium complexes. METHODS FF beads were prepared by encapsulation of hydrophobic europium complexes into the polymer layers of affinity magnetic beads using organic solvent. A sandwich immunoassay using magnetic collection of antibody-coated FF beads to a specific place was performed. Brain natriuretic peptide and prostate-specific antigen were selected as target detection antigens to demonstrate the feasibility of this approach. An immunohistochemical staining using magnetic collection of antibody-coated FF beads onto carcinoma cell samples was also performed. RESULTS The sandwich immunoassays, taking advantage of the magnetic collection of antibody-coated FF beads, detected target antigens within 5 min of sample addition. Without magnetic collection, the sandwich immunoassay using antibody-coated FF beads required long times, similar to conventional immunoassays. Using the magnetic collection of antibody-coated FF beads, immunohistochemical staining enabled discrimination of carcinoma cells within 20 min. CONCLUSIONS This proof of principle system demonstrates that immunoreactions involving the magnetic collection of antibody-coated FF beads allow acceleration of the antigen–antibody reaction. The simple magnetic collection of antibody-coated FF beads to a specific space enables rapid detection of disease-related biomarkers and identification of carcinoma cells.

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Månsson, Lisa, Carina Törn i Mona Landin-Olsson. "Islet Cell Antibodies Represent Autoimmune Response Against Several Antigens". International Journal of Experimental Diabetes Research 2, nr 2 (2001): 85–90. http://dx.doi.org/10.1155/edr.2001.85.

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To study the antigens involved in the islet cell antibody (ICA) reaction we selected 30 patient serum samples (ten in each group) positive for ICA and one other additional autoantibody, such as glutamic acid decarboxylase antibodies (GADA), thyrosine phosphatase antibodies (IA-2A) or insulin autoantibodies (IAA). The serum samples were incubated with the specific antigen (GAD65, IA-2 or insulin) and the ICA analysis and the corresponding immunoprecipitation assay were performed before and after the absorption.We could then demonstrate that specific autoantibodies against GAD65 and IA-2 could be absorbed with the corresponding antigen, since ten GADA positive and six IA-2A samples turned completely negative. However, the ICA reaction after absorption with GADA, IA-2A and insulin was still present, although at significantly lower levels. The results strongly indicate that the ICA reaction represents simultaneous autoimmunity against several other antigens beside GAD65, IA-2 and insulin.

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Kurrasch, R. H., A. V. Rutherford, M. E. Rick, M. G. Gallo, E. T. Lovelace, I. Pastan i M. C. Willingham. "Characterization of a monoclonal antibody, OVB1, which binds to a unique determinant in human ovarian carcinomas and myeloid cells." Journal of Histochemistry & Cytochemistry 37, nr 1 (styczeń 1989): 57–67. http://dx.doi.org/10.1177/37.1.2461982.

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A monoclonal antibody, OVB1, was generated against a human ovarian carcinoma cell line, OVCAR-3. The antigen reacting with this antibody was strongly expressed on the external surface of the plasma membrane of OVCAR-3 cells and cells of 4/4 other ovarian carcinoma lines. Variable density and homogeneity of expression was found on cells from 5/5 breast carcinoma lines. Various ovarian tumor specimens and normal human tissues were frozen, cryostat-sectioned, and examined for OVB1 reactivity using immunoperoxidase methods. A strong, uniform, homogeneous reaction on 10/10 ovarian carcinoma specimens and variable, non-homogeneous reactions on breast tumors were seen. Normal tissues reacting with the antibody include thyroid, pituitary pars intermedia, breast ductal epithelium, Auerbach's plexus and neuronal processes in the GI tract, colonic mucosal epithelium, and salivary gland ductal epithelium. Polymorphonuclear leukocytes, eosinophils, and approximately 13% of peripheral lymphocytes, as well as cells around germinal centers in lymph nodes and spleen, showed strong reactivity by immunofluorescence and/or immunoperoxidase. Expression of the OVB1 antigen in the myeloid cells of normal human bone marrow occurred from the promyelocyte stage through to more mature cells in a subpopulation of myeloblasts. Indirect immunofluorescence of live peripheral blood cells showed localization to the surface of PMNs, eosinophils, and certain lymphocytes. Double-immunofluorescence studies (with a direct fluorescein-anti-lactoferrin antibody conjugate) showed co-localization of OVB1 and OKM1 (anti-C3bi receptor) antibodies to specific granules of PMNs. Localization of OVB1 and OKM1 antibodies to granular structures in the PMN was confirmed by electron microscopy using the ferritin bridge technique. The antigen reacting with the OVB1 antibody was shown to be neuraminidase sensitive, but protease insensitive. The OVB1 monoclonal antibody may be useful in identification of ovarian tumors and subclassification of myeloid leukemias.

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Hong, Lisa P. T., Judith A. Scoble, Larissa Doughty, Gregory Coia i Charlotte C. Williams. "Cancer-targeting Antibody–Drug Conjugates: Site-specific Conjugation of Doxorubicin to Anti-EGFR 528 Fab' through a Polyethylene Glycol Linker". Australian Journal of Chemistry 64, nr 6 (2011): 779. http://dx.doi.org/10.1071/ch11071.

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Antibody–drug conjugates have been prepared to examine the effect that attaching small-molecule drugs to an antibody fragment has on antibody activity. The anticancer drug doxorubicin was covalently attached through a polyethylene glycol linker to a cancer-targeting, anti-epidermal growth factor receptor antibody fragment (Fab′). The reactivity of maleimide was compared with a substituted maleimide derivative (citraconimide) in conjugation reactions with cysteine residues on a Fab′. Introduction of polyethylene glycol increased aqueous solubility of the cytotoxic drug, which led to an improvement in overall yield of the conjugation reaction with the antibody fragment. Antibody–drug conjugates prepared retained activity of the parent antibody, as determined by antigen binding experiments measured by surface plasmon resonance.

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Auditore-Hargreaves, K., R. L. Houghton, N. Monji, J. H. Priest, A. S. Hoffman i R. C. Nowinski. "Phase-separation immunoassays." Clinical Chemistry 33, nr 9 (1.09.1987): 1509–16. http://dx.doi.org/10.1093/clinchem/33.9.1509.

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Abstract Solid-phase-based immunoassays have traditionally been plagued by nonspecific binding to the solid phase and by slow reaction kinetics relative to reactants that are free to diffuse in solution. We have developed two novel immunoassays in which the solid phase is generated in situ after the specific binding reaction has occurred, thereby enhancing reaction kinetics and minimizing the opportunities for non-specific binding. In the first system, the capture antibody is conjugated to an organic monomer, polymerization of which to form insoluble polymer particles is initiated by a reaction involving free radicals. The amount of signal-labeled antibody incorporated into the resulting particles is directly proportional to the concentration of antigen. The principle is illustrated for the simultaneous assay of IgG and IgM in a single sample. In the second system, capture antibody is conjugated to a polymer, the solubility of which is a function of temperature. Specific binding is conducted below the critical solution temperature of the polymer, which is then separated from solution by increasing the temperature above the critical temperature. The incorporation of signal-labeled antibody into the precipitated polymer is directly proportional to the concentration of antigen. This principle is illustrated for the assay of hepatitis B surface antigen and Chlamydia trachomatis.

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Hoffmann, Johannes J. M. L., i Willy C. M. Janssen. "HLA-B27 phenotyping with flow cytometry: further improvement by multiple monoclonal antibodies". Clinical Chemistry 43, nr 10 (1.10.1997): 1975–81. http://dx.doi.org/10.1093/clinchem/43.10.1975.

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Abstract To establish the optimal flow cytometric method for HLA-B27 phenotyping, we compared several strategies, using three monoclonal anti-B27 antibodies (from the HLA-ABC-m3, GS145.2, and FD705 clones). We used a triple-color direct immunofluorescence assay, including a T-lymphocyte-specific antibody as an internal control and an anti-HLA-Bw4 antibody. Blood samples from >400 subjects were tested. From ROC curve analysis none of the three antibodies appeared to be suitable for use as a single typing reagent. The efficiency of the test was affected by cross-reactions with other HLA antigens, notably the HLA-B7 antigen. Preincubation with anti-B7 serum efficiently inhibited this cross-reaction and raised the test efficiency considerably. We concluded that none of the anti-B27 antibodies investigated is suitable for use as a single typing reagent. Additional typing of Bw4 is not valuable, whereas inhibition of cross-reactions due to the B7 antigen will considerably improve the performance of the test. We recommend that two different monoclonal anti-B27 antibodies be used for accurate and reliable HLA-B27 phenotyping with flow cytometry.

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Gee, B., M. J. Warhol i J. Roth. "Use of anti-horseradish peroxidase antibody-gold complex in the ABC technique." Journal of Histochemistry & Cytochemistry 39, nr 6 (czerwiec 1991): 863–69. http://dx.doi.org/10.1177/39.6.1709659.

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We report a modification of the avidin-biotin-peroxidase complex (ABC) technique for the light and electron microscopic detection of antigens in tissue sections. An immunological approach was used instead of the DAB reaction to reveal ABC bound to antigen-antibody complexes. Affinity-purified polyclonal antibodies against horseradish peroxidase were complexed to particles of colloidal gold and applied for reaction with the horseradish peroxidase molecules of the ABC. For light microscopic immunolabeling, the signal produced by the anti-horseradish peroxidase antibody-gold complex required silver intensification. The ABC immunogold reaction as compared with the standard ABC technique, in particular with silver intensification of the DAB reaction product, provided superior resolution in paraffin sections. Furthermore, section pre-treatment to block endogenous peroxidase activity could be omitted and no potentially hazardous substrate was used. The ABC immunogold reaction was successfully applied for electron microscopic immunolabeling on Lowicryl K4M thin sections. We propose that the ABC immunogold reaction is a useful alternative to the standard ABC technique and can be equally well applied to light and electron microscopy.

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Sadeghimehr, Maryam, Barbara Bertisch, Francesco Negro, Maia Butsashvili, Sonjelle Shilton, Irina Tskhomelidze, Maia Tsereteli, Olivia Keiser i Janne Estill. "Hepatitis C core antigen test as an alternative for diagnosing HCV infection: mathematical model and cost-effectiveness analysis". PeerJ 9 (10.09.2021): e11895. http://dx.doi.org/10.7717/peerj.11895.

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Background The cost and complexity of the polymerase chain reaction (PCR) test are barriers to diagnosis and treatment of hepatitis C virus (HCV) infection. We investigated the cost-effectiveness of testing strategies using antigen instead of PCR testing. Methods We developed a mathematical model for HCV to estimate the number of diagnoses and cases of liver disease. We compared the following testing strategies: antibody test followed by PCR in case of positive antibody (baseline strategy); antibody test followed by HCV-antigen test (antibody-antigen); antigen test alone; PCR test alone. We conducted cost-effectiveness analyses considering either the costs of HCV testing of infected and uninfected individuals alone (A1), HCV testing and liver-related complications (A2), or all costs including HCV treatment (A3). The model was parameterized for the country of Georgia. We conducted several sensitivity analyses. Results The baseline scenario could detect 89% of infected individuals. Antibody-antigen detected 86% and antigen alone 88% of infected individuals. PCR testing alone detected 91% of the infected individuals: the remaining 9% either died or spontaneously recovered before testing. In analysis A1, the baseline strategy was not essentially more expensive than antibody-antigen. In analysis A2, strategies using PCR became cheaper than antigen-based strategies. In analysis A3, antibody-antigen was again the cheapest strategy, followed by the baseline strategy, and PCR testing alone. Conclusions Antigen testing, either following a positive antibody test or alone, performed almost as well as the current practice of HCV testing. The cost-effectiveness of these strategies depends on the inclusion of treatment costs.

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Aung, Fleur M., Virgil M. Reddy i Benjamin Lichtiger. "The Benefit of RED CELL Antigen Testing in A Transfusion Service of A Large Cancer Center". Blood 120, nr 21 (16.11.2012): 4374. http://dx.doi.org/10.1182/blood.v120.21.4374.4374.

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Abstract Abstract 4374 Introduction: DNA-based tests are increasingly being used to predict blood group phenotypes in patients who have been recently transfused to aid in alloantibody identification, to distinguish an alloantibody from an autoantibody, in patients who have received hematopoietic stem cell transplants, when RBCS are coated with immunoglobulin (+DAT) and to detect weakly expressed antigens when the patient is unlikely to make antibodies to antigen- positive RBC's transfused. The molecular bases associated with most antigens have been determined and PCR-assays available for testing RBC antigens are a powerful adjunct to the conventional hemagglutination, the gold standard technique to type RBCs for the presence or absence of blood group antigens. Purpose: This retrospective review of Red Blood cell (RBC) antigen testing data was conducted on all patients typed between January 2010 and December 2011 at MD Anderson Cancer Center. The purpose was to review the concordance between the antibody (ies) identified by serologic testing and the antigen negativity from phenotype predicted from DNA analysis for selecting antigen negative RBCs for transfusion. In addition we also wanted to review whether the extended phenotype red cell units transfused to patients with unidentified alloantibody (ies)/RACT and autoantibodies formed additional red cell alloantibody (ies) after transfusion. Methods: The technology of BioArray Solutions (Bioarray Solutions, Immuncor, Norcross, GA, USA) and the Human Erythrocyte Antigen (HEA) v1.2 BeadChip was used to assess the genotype and predicted phenotype of patients. We performed RBC antigen testing on all of our recently transfused patients with alloantibody (ies), patients whose red cell antibody panels showed reaction with all red cells tested (RACT) or unidentified antibodies, patients with positive direct antiglobulin test (DAT) and whose elution studies showing RACT or unidentified antibody (ies) and in patients with autoimmune hemolytic anemia. Results: Atotal of 912 BioArray tests were performed on 912 patients [444 (49%) males:468 (51%) females; median age 60 years (range 1–92)]. Antibody screen was positive in 424 (46%) and negative in 488 (54%) patients. A DAT was performed in 126 patients with a positive antibody screen to establish immune from non-immune hemolytic anemia. The results are listed in Tables 1,2 and 3. Conclusion: We found 100% concordance between the predicted antigen negativity phenotype and the patient's RBC alloantibody (ies) identified by serologic testing. The RBC antigen typing has allowed us to transfuse patients with unidentified antibodies and RACT with extended phenotype matched red cell units preventing the development of additional red cell alloantibody (ies) as well as allowed us to transfuse antigen-negative units preventing patient/blood donor incompatibility issues. Disclosures: No relevant conflicts of interest to declare.

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Perret, E., M. Moudjou, M. L. Geraud, J. Derancourt, M. O. Soyer-Gobillard i M. Bornens. "Identification of an HSP70-related protein associated with the centrosome from dinoflagellates to human cells". Journal of Cell Science 108, nr 2 (1.02.1995): 711–25. http://dx.doi.org/10.1242/jcs.108.2.711.

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The monoclonal antibody CTR210 raised against isolated human centrosomes strongly decorates the centrosome and more weakly a domain congruent with the Golgi apparatus in several animal cells (HeLa, 3T3, CHO, PtK2). Both decorations resist Triton extraction in conditions which totally extract the Golgi apparatus, as judged by galactosyltransferase decoration. A 67 kDa centrosomal antigen can be demonstrated in human cells with this antibody. CTR210 also decorates the centrosome or associated structures in several systems, including unicellular eukaryotes such as dinoflagellates or ciliates. A 72 kDa antigen has been identified and purified from the dinoflagellate C. cohnii and its NH2-terminal sequence partially established. It shows a close homology with HSP70 proteins. The possibility that the 72 kDa antigen belongs to this chaperone family was further supported using a mAb reacting, in most species, with HSP70. A polyclonal antibody raised against the 72 kDa antigen from C. cohnii decorates the centrosome in human cells and reacts with the CTR210 centrosomal 67 kDa antigen. These results suggest that specific chaperone proteins are associated with the centrosome in eukaryotic cells. The centrosomal chaperones could participate in the microtubule nucleation reaction or in the process of centrosome assembly.

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Ito, Hiromi, Kyoka Hoshi, Fumihiko Osuka, Mitsukazu Gotoh, Takuro Saito, Hiroshi Hojo, Rei Suzuki, Hiromasa Ohira, Takashi Honda i Yasuhiro Hashimoto. "Lectin inhibits antigen–antibody reaction in a glycoform‐specific manner: Application for detecting α2,6sialylated‐carcinoembryonic antigen". PROTEOMICS 16, nr 24 (15.09.2016): 3081–84. http://dx.doi.org/10.1002/pmic.201600117.

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Rodero, M., C. Cuéllar, T. Chivato, J. M. Mateos i R. Laguna. "Western blot antibody determination in sera from patients diagnosed with Anisakis sensitization with different antigenic fractions of Anisakis simplex purified by affinity chromatography". Journal of Helminthology 81, nr 3 (wrzesień 2007): 307–10. http://dx.doi.org/10.1017/s0022149x07820220.

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AbstractUsing Western blot techniques, the specificities of crude and purified (PAK and PAS) Anisakis simplex antigens were compared against 24 sera from patients diagnosed with Anisakis sensitization. All patients recognized a 60 kDa protein against the A. simplex crude extract, while 37.5% and 12.5% reacted with proteins of 40 and 25 kDa, respectively, when IgG was tested. In the case of IgE determination, 41.6% of sera were negative, while 12.5% and 20.8% appeared to cross-react against Toxocara canis and Ascaris suum, respectively. When the PAK antigen (A. simplex antigen purified by means of a column of IgG anti-A. simplex) was tested, immune recognition towards the 60, 40 and 25 kDa proteins increased in 83.3%, 16.7% and 4.2%, respectively, when the Ig antibodies were tested. In the case of the PAS antigen (PAK antigen purified by means of a column of IgG anti-A. suum), the reaction against the 40 and 25 kDa proteins increased to 45.8% and 25%, respectively, when Ig antibodies were used. Finally, when the EAS antigen (eluted from the anti-A. suum column after PAK purification) was tested, 83.3% of the assayed sera reacted against the 14 kDa protein, when the Ig antibodies, IgG and IgM immunoglobulins were measured. With the IgE determination, the reactions were observed in 41.7% of patients with proteins between 60 and 35 kDa against the PAS antigen. With the EAS antigen, reactive bands of 184, 84 and 14 kDa appeared. In conclusion, in the purification process of the A. simplex larval crude extract, the proteins implicated in cross-reactions with Ascaris and Toxocara were eliminated, with an important concentration of proteins responsible for the induction of specific responses.

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Grigorescu, Cristina, Liviu Ciprian Gavril, Laura Gavril, Tiberiu Lunguleac, Bogdan Mihnea Ciuntu, Delia Hinganu, Alexandru Patrascu i Paul Salahoru. "Use of Immunohistochemistry to Establish Ethiology in the Case of Primary Spontaneous Pneumotorax Patients". Revista de Chimie 69, nr 10 (15.11.2018): 2734–36. http://dx.doi.org/10.37358/rc.18.10.6614.

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Primary spontaneous pneumothorax has a complex morphopathological substrate, in which active smoking plays an essential etiopathogenic role. Inflammation of the distal airways, bronchial anomalies, perivascular eosinophilic infiltrate associated with hereditary factors and physiognomy (longilli patients) lead to obstruction of distal airwayswhich is the essential element in the emergence of emphysematous changes. Immunohistochemistry (IHC) is a technique used to identify cellular or tissue (antigens) constituents by Ag-Ac, the Ac link site being identified either by direct labeling of the antibody or by a secondary labeling method. IHC reactions are based on tissue-antibody antigen binding, the latter being evidenced by direct conjugation to tracer molecules (direct reaction) or by another chain of other labeled free antibody linkages. We can consider the immunohistochemical method as having a potential utility, especially in selected patients, where there are sufficient clinical and epidemiological reasons to suspect a pneumothorax-causing disease but where the classical investigations did not provide diagnostic performance.

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Nurhadi, Nurhadi, Kamaruzaman Sijam i Inon Sulaiman. "PRODUCTION OF POLYCLONAL ANTIBODY TO THE COAT PROTEIN OF CITRUS TRISTEZA VIRUS IN CHICKEN EGGS". Indonesian Journal of Agricultural Science 4, nr 1 (25.10.2016): 18. http://dx.doi.org/10.21082/ijas.v4n1.2003.18-26.

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Citrus tristeza virus (CTV) is one of the most destructive diseases in many citrus growing areas of Indonesia. Effective strategies for controlling CTV depend on diagnostic procedure namely enzyme-linked immunosorbent assay (ELISA). Study aimed to purify the CTV antigen and produced its polyclonal antibody. Virion of the severe CTV isolate designated UPM/ T-002 was concentrated by polyethylene glycol (PEG) precipitation combined with low speed centrifugation. Semipurified antigen was further purified by sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE). The specific coat protein (CP) band of CTV with molecular weight of 25 kD was excised and eluted using elution buffer containing 0.25 M Tris-HCl pH 6.8 + 0.1% SDS, then used as antigen for injection into 6-month-old female of White Leghorn chicken. Results, showed than the specific polyclonal antibody raised against the 25-kDa CP had a titer of approximately 104, gave low background reaction with healthy plant sap and reacted specifically with CTV isolates. The reaction was equally strong for a severe, a moderate, a mild, and a symptomless isolate, suggesting a broad reaction range of this antibody toward different CTV isolates. Optimal virus titer can be obtained since virus loss during purification could be minimized and the highly purified antigen as an immunogen could be obtained by cutting out the CP band from SDS-PAGE gels. Large amount of highly titer of CTV antibody can be produced in chicken egg. The simplicity of the procedure makes it economically acceptable and technically adoptable because the antibody can be produced in basic laboratory.

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Nurhadi, Nurhadi, Kamaruzaman Sijam i Inon Sulaiman. "PRODUCTION OF POLYCLONAL ANTIBODY TO THE COAT PROTEIN OF CITRUS TRISTEZA VIRUS IN CHICKEN EGGS". Indonesian Journal of Agricultural Science 4, nr 1 (25.10.2016): 18. http://dx.doi.org/10.21082/ijas.v4n1.2003.p18-26.

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Citrus tristeza virus (CTV) is one of the most destructive diseases in many citrus growing areas of Indonesia. Effective strategies for controlling CTV depend on diagnostic procedure namely enzyme-linked immunosorbent assay (ELISA). Study aimed to purify the CTV antigen and produced its polyclonal antibody. Virion of the severe CTV isolate designated UPM/ T-002 was concentrated by polyethylene glycol (PEG) precipitation combined with low speed centrifugation. Semipurified antigen was further purified by sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE). The specific coat protein (CP) band of CTV with molecular weight of 25 kD was excised and eluted using elution buffer containing 0.25 M Tris-HCl pH 6.8 + 0.1% SDS, then used as antigen for injection into 6-month-old female of White Leghorn chicken. Results, showed than the specific polyclonal antibody raised against the 25-kDa CP had a titer of approximately 104, gave low background reaction with healthy plant sap and reacted specifically with CTV isolates. The reaction was equally strong for a severe, a moderate, a mild, and a symptomless isolate, suggesting a broad reaction range of this antibody toward different CTV isolates. Optimal virus titer can be obtained since virus loss during purification could be minimized and the highly purified antigen as an immunogen could be obtained by cutting out the CP band from SDS-PAGE gels. Large amount of highly titer of CTV antibody can be produced in chicken egg. The simplicity of the procedure makes it economically acceptable and technically adoptable because the antibody can be produced in basic laboratory.

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