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Journal articles on the topic 'ELISA en sandwich'

Author: Grafiati

Published: 11 September 2021

Last updated: 14 February 2022

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1

Katsurada, Akemi, Yoshiaki Hagiwara, Kazuya Miyashita, Ryousuke Satou, Kayoko Miyata, Naro Ohashi, L. Gabriel Navar, and Hiroyuki Kobori. "Novel sandwich ELISA for human angiotensinogen." American Journal of Physiology-Renal Physiology 293, no. 3 (September 2007): F956—F960. http://dx.doi.org/10.1152/ajprenal.00090.2007.

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We recently reported that urinary excretion rates of angiotensinogen (UAGT) provide a specific index of intrarenal renin-angiotensin (ANG) system (RAS) status in ANG II-dependent hypertensive rats. When this is shown to be applicable to human subjects, a diagnostic test to identify those hypertensive patients most likely to respond to an RAS blockade could provide useful information to allow a mechanistic rationale for selection of an optimized approach to treatment of hypertensive patients. However, simple and accurate methods to measure human angiotensinogen (hAGT) are unavailable. For future studies of human subjects, we developed antibodies and a sensitive and specific quantification system for hAGT using a sandwich ELISA. We raised two antibodies against hAGT: a mouse monoclonal antibody and a rabbit polyclonal antibody. The standard curve of this ELISA exhibited a high linearity (0.31–20 ng/ml). The correlation coefficient was >0.99. Plasma angiotensinogen concentrations of healthy volunteers ranged from 28 to 71 μg/ml ( n = 10). The ratio of UAGT to urinary creatinine concentration ranged from 5.0 to 30 μg/g ( n = 7). Intra- and interassay coefficients of variation ranged from 4.4 to 5.5% and from 4.3 to 7.0%, respectively. This ELISA system had no cross-reactivity with major proteins in proteinuric urine samples, such as human albumin, immunoglobulin, or transferrin. Moreover, the cross-reactivity of the system with angiotensin peptides was also negligible. This hAGT ELISA will be a useful tool to investigate the relationship of UAGT and reactivity to antihypertensive drugs in hypertensive patients.

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2

Zhao, Weilin, Seinyone Pao, Fatima Malik, James Soh, Sonalis Fernandez, and William J. Chirico. "A Sandwich ELISA for Phosphoglycerate Kinase." Journal of Immunoassay and Immunochemistry 29, no. 3 (June 2, 2008): 220–33. http://dx.doi.org/10.1080/15321810802119588.

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3

Ibrahim R B Aly, Samir A M Zaahkouk, Alaa A M Samn, Ibrahim A E Zahran, and Sawsan A M EL-Shamy. "Effective diagnosis of schistosomiasis haematobium by Silver beads Sandwich ELISA." International Journal of Research in Pharmaceutical Sciences 11, SPL4 (December 21, 2020): 1398–404. http://dx.doi.org/10.26452/ijrps.v11ispl4.4312.

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Schistosomiasis is a major public health problem. Diagnosis by simple and rapid immunoassays is a priority. The magnetic bead immunoassay using magnetic nanoparticles conjugated with anti-schistosomal antibody was evaluated for diagnosing human schistosomiasis infection. The present study was to evaluate the sandwich ELISA as a simple test for the detection of schistosomal antigen (CSA) in serum and urine samples of S. haematobium patients and compare it with ELISA. Investigation conducted on eighty six cases divided to three groups, 34 were positively for Schistosoma haematobium, 32 were positively for intestinal parasites ova and negative for S. haematobium ova in urine and 20 were negative urine and stool examination (Control). Immunomagnetical bead based Enzyme-linked immunosorbent assay (ELISA) using for detected for antigen in sera and urine infected by S. haematobium. Sandwich ELISA sensitivities was 79.4% (serum) and 73.5% (urine) and which increase by used nano-sandwich ELISA to 88.2% (serum) and 82.4% (urine), respectively. Sandwich ELISA specificities was 86.4% (serum) and 80.8% (urine) and which increase by used nano-sandwich ELISA to 93.3% (serum) and 88.5% (urine). We found that, nano-sandwich ELISA assay had highly sensitive and specifically and technical method was applicably, fast, cheaper, accurate and promising diagnostic method for schistosomiasis.

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Rhee, Jong Il, Jörg Hagedorn, Thomas Scheper, and Karl Schügerl. "Flow-injection sandwich ELISA for bioprocess monitoring." Journal of Automated Methods and Management in Chemistry 21, no. 4 (1999): 121–25. http://dx.doi.org/10.1155/s1463924699000140.

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A fully automated flow-injection immunoassay based on sandwich enzyme-linked immunosorbent assay (ELISA) is described for the model system: protein G-sepharose, rabbit IgG and horseradish peroxidase (HRP)-labelled protein A. After injecting rabbit IgG and HRP-labelled protein A into a cartridge containing protein G-sepharose sequentially, a mixture of hydrogen peroxide and the redox indicator, 2.2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) is passed through the cartridge. The HRP-labelled protein A bound in the cartridge is directly proportional to the concentration of rabbit IgG. The colour variation of ABTS caused during the reaction between HRP and H202in the cartridge is detected photometrically. The whole assay procedure is controlled and evaluated by a computer. Rabbit IgG and HRP-labelled protein A are also detected by a fluorometer, which is introduced into the flow system. In the flow-injection sandwich ELISA, the slope of the calibration curve is 0.4491 in the range of 0 and 300 μg ml-1rabbit IgG, while it is 0.1274 in the heterogeneous immunoassay. So the flow-injection sandwich ELISA system is found to be more sensitive than a heterogeneous immunoassay for the monitoring of the model protein.

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5

Huang, Ruo-Pan, Ying Lin, Li-Pai Chen, Weimin Yang, and Ruochun Huang. "ELISA-based Protein Arrays: Multiplexed Sandwich Immunoassays." Current Proteomics 1, no. 3 (July 1, 2004): 199–210. http://dx.doi.org/10.2174/1570164043379226.

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6

Oka, T., M. Ito, T. Egashira, N. E. Miller, and H. Hattori. "Plasma PLTP concentration measured by sandwich ELISA." Atherosclerosis 151, no. 1 (July 2000): 220. http://dx.doi.org/10.1016/s0021-9150(00)81001-1.

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7

Rhee, Jong Il, Jorg Hagedorn, Thomas Scheper, and Karl Schugerl. "Flow-injection sandwich ELISA for bioprocess monitoring." Journal of Automated Methods & Management in Chemistry 21, no. 4 (July 1, 1999): 121–25. http://dx.doi.org/10.1080/146392499300613.

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8

Lim, Yoon, Jin-Hua Zhong, and Xin-Fu Zhou. "Development of mature BDNF-specific sandwich ELISA." Journal of Neurochemistry 134, no. 1 (April 21, 2015): 75–85. http://dx.doi.org/10.1111/jnc.13108.

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9

Panchal, Manoj, K. Muralidhar, and S. K. Gupta. "A Sensitive Sandwich ELISA for Buffalo Prolactin." Journal of Immunoassay and Immunochemistry 28, no. 2 (March 22, 2007): 147–54. http://dx.doi.org/10.1080/15321810701212062.

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10

VANREE, R., S. STAPEL, and R. AALBERSE. "Reverse-sandwich ELISA may underestimate antibody titers." Journal of Allergy and Clinical Immunology 84, no. 4 (October 1989): 562–63. http://dx.doi.org/10.1016/0091-6749(89)90371-0.

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11

NUNTAPRASERT, A., Y. MORI, K. TSUKIYAMAKOHARA, and C. KAI. "Establishment of swine interleukin-6 sandwich ELISA." Comparative Immunology, Microbiology and Infectious Diseases 28, no. 2 (March 2005): 121–30. http://dx.doi.org/10.1016/j.cimid.2004.08.003.

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12

Troutt, Jason S., Robert W. Siegel, Jinbiao Chen, John H. Sloan, Mark A. Deeg, Guoqing Cao, and Robert J. Konrad. "Dual-Monoclonal, Sandwich Immunoassay Specific for Glucose-Dependent Insulinotropic Peptide1-42, the Active Form of the Incretin Hormone." Clinical Chemistry 57, no. 6 (June 1, 2011): 849–55. http://dx.doi.org/10.1373/clinchem.2010.159954.

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BACKGROUND Glucose-dependent insulinotropic peptide (GIP) is an incretin peptide secreted by intestinal K cells that stimulates insulin secretion in a glucose-dependent manner. It is secreted as an active, intact 42–amino acid peptide GIP1-42, which is rapidly degraded by dipeptidyl peptidase 4 to GIP3-42, which is inactive. There is currently no described monoclonal antibody–based sandwich immunoassay to quantify concentrations of GIP1-42, the active form of the peptide. METHODS To create a sandwich ELISA for GIP1-42, we generated a monoclonal antibody specific for the intact N-terminus of the peptide, which was further optimized to increase its affinity. We used this antibody as a conjugate antibody in a sandwich ELISA and paired it with an anti–total GIP capture monoclonal antibody to create a dual monoclonal sandwich ELISA for GIP1-42. RESULTS The sandwich ELISA was highly specific for GIP1-42 and did not recognize GIP3-42. The ELISA demonstrated a broad dynamic range and a lower limit of quantification of 5 ng/L. Using the ELISA, we were able to show that GIP1-42 concentrations in healthy volunteers increased dramatically in the postprandial state compared to the fasting state. GIP1-42 values were correlated with total GIP values overall; however, there was substantial interindividual variation. CONCLUSIONS The use of an N-terminal–specific monoclonal antibody in a sandwich ELISA format provides a robust and convenient method for measuring concentrations of GIP1-42, the active form of the incretin hormone. This ELISA should help to improve our understanding of the role of GIP1-42 in regulating glucose-dependent insulin secretion.

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13

Zhang, Yuan, Gang Xu, Lu Zhang, Jiakai Zhao, Pinpin Ji, Yaning Li, Baoyuan Liu, et al. "Development of a double monoclonal antibody–based sandwich enzyme-linked immunosorbent assay for detecting canine distemper virus." Applied Microbiology and Biotechnology 104, no. 24 (November 7, 2020): 10725–35. http://dx.doi.org/10.1007/s00253-020-10997-y.

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Abstract Canine distemper virus (CDV) infection causes mass mortality in diverse carnivore species. For effective virus surveillance, rapid and sensitive assays are needed to detect CDV in field samples. In this study, after BABL/c mice were immunized with recombinant CDV-fusion (F) protein, monoclonal antibodies (mAbs) against recombinant CDV-F protein (designated 1A5, 1A6, and 7D5) were produced using traditional hybridoma cell technology. Next, capture antibody (1A6, 800 ng/well) and horseradish peroxidase (HRP)–conjugated detection antibody (HRP-7D5, 1:100, 500 ng/well) were used in a double monoclonal antibody–based sandwich enzyme-linked immunosorbent assay (ELISA) for CDV detection after optimization of both mAb amounts per well using a checkerboard titration test. Based on sandwich ELISA test results for 120 known CDV-negative samples, the cutoff value for a positive result was set to an OD450 nm value ≥ 0.196. As compared with test results obtained from commercial immune colloidal gold test strips, the low limits of detection for the two assays were revealed to be 100 TCID50 per 100 μL. In addition, the sandwich ELISA agreed 100% and 96.4% with commercial immune colloidal gold test strips when testing serum and stool samples. The sandwich ELISA assay provided statistically similar CDV detection. Thus, the sandwich ELISA developed here to detect CDV in fecal and serum samples provided good sensitivity, high specificity, and good reproducibility and should serve as an ideal method for large-scale surveillance of CDV infections in carnivores. Key points • Three CDV mAbs that recognized different epitopes and bound to virion were generated. • The sandwich ELISA based mAbs to detect CDV in fecal and serum samples was developed. • The sandwich ELISA is an ideal method for detecting CDV infections in the field.

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14

Ge, Meng, Wei Luo, Daliang Jiang, Runcheng Li, Wenwei Zhao, Guoliang Chen, Xingdong Yang, and Xinglong Yu. "Development and Application of a Double-Antigen Sandwich Enzyme-Linked Immunosorbent Assay for Detection of Antibodies to Porcine Circovirus 2." Clinical and Vaccine Immunology 19, no. 9 (July 18, 2012): 1480–86. http://dx.doi.org/10.1128/cvi.00234-12.

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ABSTRACTA double-antigen sandwich enzyme-linked immunosorbent assay (ELISA) is described for detection of porcine circovirus 2 (PCV2) antibodies using the well-characterized recombinant PCV2 capsid protein. In a comparative test of 394 pig sera against an indirect immunofluorescence (IIF) test and a commercial ELISA kit (also based on the recombinant PCV2 capsid protein), the results showed that the diagnostic sensitivity, specificity, and accuracy of the assay were, respectively, 90.61, 94.02, and 91.62% compared with IIF and 94.38, 95.28, and 94.67% compared with the commercial ELISA kit. Assay of 12 PCV-free pigs over a 5-week period produced only PCV2-negative titers by all 3 methods. These results and the seroprofiles of 4 pig farms obtained by both the commercial ELISA kit and the double-antigen sandwich ELISA indicate that the sandwich ELISA is a reliable method for detection of antibodies to PCV2. Additionally, the method described here permits the use of undiluted test serum samples simultaneously loaded with horseradish peroxidase (HRP)-conjugated antigen into the test well, and the complete test procedure can be performed in less than 90 min. This double-antigen sandwich ELISA should be a useful tool to aid swine industry professionals in deciding the intervention strategies for the control of PCV2-associated diseases.

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15

Graham, Nicholas A., Melissa D. Pope, Tharathorn Rimchala, Beijing K. Huang, and Anand R. Asthagiri. "A Microtiter Assay for Quantifying Protein-Protein Interactions Associated with Cell-Cell Adhesion." Journal of Biomolecular Screening 12, no. 5 (August 2007): 683–93. http://dx.doi.org/10.1177/1087057107301941.

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Cell-cell adhesions are a hallmark of epithelial tissues, and the disruption of these contacts plays a critical role in both the early and late stages of oncogenesis. The interaction between the transmembrane protein E-cadherin and the intracellular protein β-catenin plays a crucial role in the formation and maintenance of epithelial cell-cell contacts and is known to be downregulated in many cancers. The authors have developed a protein complex enzyme-linked immunosorbent assay (ELISA) that can quantify the amount of β-catenin bound to E-cadherin in unpurified whole-cell lysates with a Z′ factor of 0.74. The quantitative nature of the E-cadherin:β-catenin ELISA represents a dramatic improvement over the low-throughput assays currently used to characterize endogenous E-cadherin:β-catenin complexes. In addition, the protein complex ELISA format is compatible with standard sandwich ELISAs for parallel measurements of total levels of endogenous E-cadherin and β-catenin. In 2 case studies closely related to cancer cell biology, the authors use the protein complex ELISA and traditional sandwich ELISAs to provide a detailed, quantitative picture of the molecular changes occurring within adherens junctions in vivo. Because the E-cadherin: β-catenin protein complex plays a crucial role in oncogenesis, this protein complex ELISA may prove to be a valuable quantitative prognostic marker of tumor progression. ( Journal of Biomolecular Screening 2007:683-693)

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16

Koppelman, Stef J., Ashley L. Lardizabal, Lynn Niemann, Joe L. Baumert, and Steve L. Taylor. "Development of a Sandwich Enzyme-Linked Immunosorbent Assay for Detection and Quantification of Clam Residues in Food Products." BioMed Research International 2021 (March 19, 2021): 1–9. http://dx.doi.org/10.1155/2021/6685575.

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Seafood is a frequent cause of allergic reactions to food globally. The presence of undeclared trace amounts of clam can cause allergic reactions in sensitive individuals. Limited tools are available to test food products for the presence of traces of clam. We report on the development of a sandwich ELISA that can detect and quantify clam protein in food. Antisera against a mix of two commercially important clam species, Atlantic Surf (Spisula solidissima) and ocean quahog (Arctica islandica), were raised in rabbit and sheep. A sandwich ELISA was constructed with this antisera, and sensitivity and specificity were evaluated. Also, model food products spiked with clam protein were analyzed to assess the performance of the ELISA. Comparison was made with a commercially available ELISA for crustacea. The lower limit of quantification of the sandwich ELISA is 2.5 ppm clam protein in food samples, allowing the detection of low amounts of clam that may trigger a reaction in clam allergic patients. The sandwich ELISA was highly specific with cross-reactivity only noted for other molluscan shellfish (mussel and scallop). Clam protein in tomato juice and potato cream soup was detected well with recoveries ranging from 65 to 74% and from 74 to 113%, respectively. However when potato cream soup was retorted, the recover fell to 20%, imposing the risk of underestimating the clam content of a food product. A commercially available crustacean ELISA test was not suitable to detect clam protein. The sandwich ELISA described here is suitable for detection and quantification of clam protein in food products. Care should be taken with food products that have been retorted as the results may be underestimated.

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17

MIYAMOTO, TAKAHISA, HUAIZE TIAN, KIYOSHI MATSUNO, RYOJI TAKATA, and SHOJI HATANO. "Application of Monoclonal Antibodies to Dulcitol 1-Phosphate Dehydrogenase for Rapid Detection of Salmonella." Journal of Food Protection 58, no. 8 (August 1, 1995): 847–52. http://dx.doi.org/10.4315/0362-028x-58.8.847.

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Monoclonal antibodies raised against dulcitol 1-phosphate dehydrogenase of Salmonella typhimurium IFO 12529 were screened against 20 serotypes of Salmonella and 13 non-Salmonella bacteria. A sandwich-capture, enzyme-linked immunosorbent assay (sandwich ELISA) was developed for detection of Salmonella in food. The assay utilizes two monoclonal antibodies (DUI2 and DU28) which show no cross-reactions with non-Salmonella bacteria. The limit of detection of the sandwich ELISA was about 1 × 107 CPU/ml. After cultivation in a medium containing dulcitol at 37°C for 18 h followed by the sandwich ELISA. 1 CPU of Salmonella was detected. Although a high inoculum level of E. coli interfered with the detection of Salmonella, the interference was minimized by using a selective dulcitol-magnesium chloride-pyridinesulfonic acid medium for enrichment. The novel ELISA procedure detected Salmonella in chicken filtrates inoculated with 1.4 CPU/50 m1 and 1.3 × 107 CPU/50 ml of E. coli within 25 h.

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Allam, G., I. R. Bauomy, Z. M. Hemyeda, T. M. Diab, and T. F. Sakran. "Diagnostic potential ofFasciola gigantica-derived 14.5 kDa fatty acid binding protein in the immunodiagnosis of bubaline fascioliasis." Journal of Helminthology 87, no. 2 (March 13, 2012): 147–53. http://dx.doi.org/10.1017/s0022149x12000168.

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AbstractThe 14.5 kDa fatty acid binding protein (FABP) was isolated from the crude extract of adultFasciola giganticaworms. Polyclonal anti-FABP IgG was generated in rabbits immunized with prepared FABP antigen. Sandwich enzyme-linked immunosorbent assay (ELISA) was applied to detect coproantigen in stools and circulatingFasciolaantigen (CA) in sera of 126 water buffaloes by using purified and horseradish peroxidase (HRP)-conjugated anti-FABP IgG. Sandwich ELISA sensitivity was 96.97% and 94.95%; while specificity was 94.12% and 82.35% for coproantigen and CA detection, respectively. However, sensitivity and specificity of the Kato–Katz technique was 73.74% and 100%, respectively. The diagnostic efficacy of sandwich ELISA was 96.55% and 93.1% for coproantigen and CA detection, respectively. In contrast, the diagnostic efficacy of the Kato–Katz technique was 77.59%. In conclusion, these results demonstrate that the purified 14.5 kDa FABP provides a more suitable antigen for immunodiagnosis of early and current bubaline fascioliasis by using sandwich ELISA.

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Amosova, I. V., and M. P. Grudinin. "Diagnostic Potential of Monoclonal Antibodies to Adenovirus." Biotekhnologiya 37, no. 2 (2021): 48–53. http://dx.doi.org/10.21519/0234-2758-2021-37-2-48-53.

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The diagnostic potential of 4B7 and 6B12 monoclonal antibodies (mAbs) against human adenovirus hexon protein has been studied in various immunological tests, namely: in-cell enzyme-linked immunosorbent assay (cell-ELISA), sandwich ELISA, and immunofluorescent assay (IFA). It was shown that the sensitivity of cell-ELISA, sandwich ELISA and IFA was 96%, 86% and 84%, respectively as compared to PCR. Thus, 4B7 and 6B12 mAbs are promising immunoreagents for the construction of various diagnostic kits to use in laboratory practice for adenovirus detection in clinical samples. monoclonal antibodies, human adenovirus, adenovirus infection, diagnosis

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20

Miyazawa, H., H. Bannai, T. Yanase, C. Morita, S. Satoh, J. Sugiyama, S. Taniguchi, and S. Inouye. "A Reverse-Sandwich Enzyme-Linked Immunosorbent Assay for Verocytotoxin 1 and 2 Antibodies in Human and Bovine Sera." Clinical Diagnostic Laboratory Immunology 6, no. 5 (September 1, 1999): 701–4. http://dx.doi.org/10.1128/cdli.6.5.701-704.1999.

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ABSTRACT A reverse-sandwich enzyme-linked immunosorbent assay (ELISA), in which an antibody is sandwiched by antigens, was established for the titration of antibodies to verocytotoxins (VT) in human and animal sera. This assay has two advantages over a conventional indirect ELISA: (i) higher specificity and sensitivity and (ii) the ability to comparably titrate antibodies from different species. The VT1 (Shiga-like toxin 1) antibody-positive rates were 5% in 202 normal adult humans and 99% in 93 normal cattle at a dairy farm. This ELISA is most suitable for seroepidemiologic studies of infections with VT-producing Escherichia coli in humans and various animal species.

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PANDA, RAKHI, HANS F. ZOERB, CHUNG Y. CHO, LAUREN S. JACKSON, and ERIC A. E. GARBER. "Detection and Quantification of Gluten during the Brewing and Fermentation of Beer Using Antibody-Based Technologies." Journal of Food Protection 78, no. 6 (June 1, 2015): 1167–77. http://dx.doi.org/10.4315/0362-028x.jfp-14-546.

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In 2013 the U.S. Food and Drug Administration (FDA) defined the term “gluten-free” and identified a gap in the analytical methodology for detection and quantification of gluten in foods subjected to fermentation and hydrolysis. To ascertain the ability of current enzyme-linked immunosorbent assays (ELISAs) to detect and quantify gluten in fermented and hydrolyzed products, sorghum beer was spiked in the initial phases of production with 0, 20, and 200 μg/ml wheat gluten, and samples were collected throughout the beer production process. The samples were analyzed using five sandwich ELISAs and two competitive ELISAs and by sodium dodecyl sulfate–polyacrylamide gel electrophoresis with Western analysis employing four antibodies (MIoBS, R5, G12, and Skerritt). The sensitivity of the MIoBS ELISA (0.25 ppm) enabled the reliable detection of gluten throughout the manufacturing process, including fermentation, when the initial concentration of 20 μg/ml dropped to 2 μg/ml. The R5 antibody–based and G12 antibody–based sandwich ELISAs were unable to reliably detect gluten, initially at 20 μg/ml, after the onset of production. The Skerritt antibody–based sandwich ELISA overestimated the gluten concentration in all samples. The R5 antibody–based and G12 antibody–based competitive ELISAs were less sensitive than the sandwich ELISAs and did not provide accurate results for quantifying gluten concentration. The Western analyses were able to detect gluten at less than 5 μg/ml in the samples and confirmed the results of the ELISAs. Although further research is necessary before all problems associated with detection and quantification of hydrolyzed and fermented gluten are resolved, the analytical methods recommended by the FDA for regulatory samples can detect ≥20 μg/ml gluten that has undergone brewing and fermentation processes associated with the manufacture of beer.

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SHIRASU, Naoto, Takeshi ONODERA, Kazutaka NAGATOMO, Yasuyuki SHIMOHIGASHI, Kiyoshi TOKO, and Kiyoshi MATSUMOTO. "Noncompetitive Immunodetection of Benzaldehyde by Open Sandwich ELISA." Analytical Sciences 25, no. 9 (2009): 1095–100. http://dx.doi.org/10.2116/analsci.25.1095.

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23

Saika, Y., S. Yamashita, N. Sakai, K. Hirano, Y. Utida, S. Itou, and Y. Matsuzawa. "Application of a sandwich ELISA for apo B48." Atherosclerosis 151, no. 1 (July 2000): 161. http://dx.doi.org/10.1016/s0021-9150(00)80732-7.

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Kummitha, China Malakondaiah, Kristine M. Mayle, Mark A. Christman, Sudhir P. Deosarkar, Anthony L. Schwartz, Kelly D. McCall, Leonard D. Kohn, Ramiro Malgor, and Douglas J. Goetz. "A sandwich ELISA for the detection of Wnt5a." Journal of Immunological Methods 352, no. 1-2 (January 2010): 38–44. http://dx.doi.org/10.1016/j.jim.2009.11.005.

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Hoffmann, Louis G., A. Kay Pitts, Peter Densen, John M. Weiler, John E. Butler, and Jeffrey H. Peterman. "A sandwich ELISA for properdin in clinical specimens." Journal of Immunological Methods 98, no. 2 (April 1987): 161–72. http://dx.doi.org/10.1016/0022-1759(87)90001-9.

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Saha, PK, MH Ali, MB Rahman, and MA Islam. "DETERMINATION OF SENSITIVITY AND SPECIFICITY OF IN-HOUSE SANDWICH ELISA FOR THE DETECTION OF INFECTIOUS BURSAL DISEASE VIRUSES." Bangladesh Journal of Veterinary Medicine 8, no. 2 (July 11, 2012): 97–106. http://dx.doi.org/10.3329/bjvm.v8i2.11185.

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The study was designed for the development of an In-House sandwich ELISA as a suitable serological method for the rapid detection of infectious bursal disease virus (IBDV). The test was also designed to compare and evaluate its sensitivity and specificity with other traditional methods used for the detection of IBDV from field outbreak cases prevalent among the poultry population of Bangladesh. To develop the In-House sandwich ELISA, hyper-immune serum was raised against live IBDV vaccine in rabbit which was used to coat each of the 96-well flat bottomed polystyrene microtitre plate whereas, hyper-immune sera raised in chickens against IBDV used as secondary antibody. The newly developed In-House sandwich ELISA was standardized by dispensing different dilutions (10-1 up to 10-4) of rabbit serum. Among them, the 10-2 dilution of serum showed most suitable reading for the detection of IBD virus and used to coat the plate to evaluate its sensitivity and specificity. Sensitivity test was done by different dilutions (10-0 to 10-4) of reference IBD virus. The virus dilution, 10-3 was the highest dilution having lowest capacity to bind with coated antibody of the ELISA plate which indicated that IBD viruses was absent in the dilutions of above 10-3. The cut-off value of negative control samples was determined as 0.937 which indicated titer of tested samples >0.937 was positive and <0.937 was negative. Specificity test was performed using different known viruses (IBDV and NDV) using different dilutions (10-1 up to 10-4). Only the IBDV showed positive result which indicated high specificity of newly developed ELISA plate. A total of 26 samples (feces, cloacal swab, spleen and bursa) from control group, experimental and natural IBDV outbreaks were used as field viral antigen for the evaluation of sensitivity and specificity of the newly developed In-House sandwich ELISA. In case of experimental infection, 5 (62.5%) of 8 feces sample but none of cloacal swab were positive for IBDV whereas, all bursa and spleen samples were positive by both InHouse sandwich ELISA and AGIDT. In case of natural outbreak cases, 6 of 6 bursal samples and 4 of 6 spleen samples were positive by In-House sandwich ELISA whereas, AGIDT detected all bursal and 3 spleen samples. No virus was detected from the samples of control group. The result showed 92.85% specificity of the developed sandwich ELISA for detection of IBDV with AGIDT which indicated that the developed ELISA is a sensitive, specific, cost effective and reliable tool for the detection of IBDV antigen from a large number of field samples.DOI = http://dx.doi.org/10.3329/bjvm.v8i2.11185 Bangl. J. Vet. Med. (2010). 8 (2) : 97-106

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27

Speirs, Joan I., and Mumtaz Akhtar. "Detection of Escherichia coli cytotoxins by enzyme-linked immunosorbent assays." Canadian Journal of Microbiology 37, no. 8 (August 1, 1991): 650–53. http://dx.doi.org/10.1139/m91-110.

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Sandwich enzyme-linked immunosorbent assays (ELISAs) were developed to detect Escherichia coli cytotoxins. Wells were coated with monoclonal antibodies from hybridomas 13C4 and (or) 11E10, and biotin conjugates of these antibodies were used for detecting verotoxin 1 and Shiga-like toxin II, respectively. Sensitivities were about 100 and 200 cytotoxic doses, respectively. Verotoxin 2 was detected by ELISA with monoclonal antibody 11E10, but at a sensitivity of only about 4000 cytotoxic doses. ELISA results of polymyxin-treated cell extracts from cultures of 67 E. coli strains were in agreement with Vero cell assay as regards the presence and type of toxin. Key words: Escherichia coli, cytotoxin, ELISA.

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Arvedson, Tara L., George Doellgast, Hossein Salimi-Moosavi, Chadwick King, Ian Foltz, Ching Chen, Hongyan Li, and Barbra J. Sasu. "Development of a Sandwich ELISA to Detect Hepcidin in Human Serum." Blood 114, no. 22 (November 20, 2009): 2000. http://dx.doi.org/10.1182/blood.v114.22.2000.2000.

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Abstract Abstract 2000 Poster Board I-1022 Hepcidin is a 25 amino acid peptide that is the central mediator of iron metabolism. Iron excess, deficiency and maldistribution have been implicated in the etiology of many diseases including atherosclerosis, diabetes, neurodegeneration and the anemia of inflammation. Determination of hepcidin levels may be useful in diagnosis and treatment decisions for some or all of these diseases. Serum hepcidin measurement has so far been limited to a prohepcidin (60 amino acid hepcidin precursor) ELISA, mass spectrometry (MS)-based assays or competition ELISAs using polyclonal anti-hepcidin antibodies. The current work describes the generation of a sandwich ELISA using monoclonal antibodies to detect human hepcidin (hHepc) and optimization of assay conditions to resolve inconsistencies between MS- and ELISA-based detection. The ability of two anti-hHepc antibodies to sandwich (bind simultaneously) with hHepc was demonstrated using a rabbit polyclonal antibody preparation from hHepc-immunized animals. The same polyclonal antibody preparation was used for both hHepc capture and detection. The limit of detection achieved with this assay was O.D.450<1, suggesting that only a small proportion of the total antibodies could bind concurrently. To improve hHepc detection, a panel of monoclonal antibodies was screened for the ability to sandwich. Antibody epitope characterization studies using purified antibodies and >1000 hybridoma supernatants identified three classes of antibodies: classes 1 and 2 each recognized epitopes found in both full length mature hHepc (hHepc 25) and a shorter version (hHepc 20); class 3 bound a different epitope and demonstrated an increased affinity for hHepc 25 over hHepc 20. The majority of antibodies characterized were in class 1 while antibodies in classes 2 and 3 were rare (∼1% of antibody panel) highlighting the difficulty in achieving a sandwiching event. Antibodies 19D12 (class 1) and 23F11 (class 2) were identified as the optimal sandwich pair with a detection range of approximately 0.2-1000 ng/ml using synthetic hHepc. Initial comparisons of data generated using the sandwich ELISA and a fully-quantitative MS-based assay demonstrated a lack of consistent agreement. This issue was somewhat addressed by introduction of an alkaline treatment step to dissociate any protein/hHepc complexes in serum. Subsequent comparison of the two assays using sera from several different patient populations (anemia of cancer, chemotherapy-induced anemia, kidney disease) as well as healthy donors demonstrated good correlation (R2 range = 0.83-0.92; n=237). This sandwich ELISA may represent a tool for aligning the MS and ELISA-generated results in a format that has the potential to be high throughput and widely available. Disclosures: Arvedson: Amgen: Employment. Doellgast:Amgen: Employment. Salimi-Moosavi:Amgen: Employment. King:Amgen: Employment. Foltz:Amgen: Employment. Chen:Amgen: Employment. Li:Amgen: Employment. Sasu:Amgen: Employment.

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Butterfield, Anthony M., Peng Luan, Derrick R. Witcher, Joseph Manetta, Anthony T. Murphy, Victor J. Wroblewski, and Robert J. Konrad. "A Dual-Monoclonal Sandwich ELISA Specific for Hepcidin-25." Clinical Chemistry 56, no. 11 (November 1, 2010): 1725–32. http://dx.doi.org/10.1373/clinchem.2010.151522.

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BACKGROUND Hepcidin, a key regulator of iron metabolism, binds to the iron transporter ferroportin to cause its degradation. In humans, hepcidin deficiency has been linked to hemochromatosis and iron overload, whereas increased concentrations have been reported in anemia of cancer and chronic disease. There is currently an unmet clinical need for a specific immunoassay with a low limit of quantification to measure serum concentrations of hepcidin-25, the active form of the protein. METHODS We generated 2 antihepcidin-25 monoclonal antibodies and used them to build a sandwich ELISA. We correlated ELISA results to hepcidin-25 measurements by LC-MS and used ELISA to measure serum hepcidin-25 concentrations in normal individuals, cancer patients, and patients with rheumatoid arthritis. RESULTS The sandwich ELISA was highly specific for hepcidin-25, having a limit of quantification of 0.01 μg/L (10 pg/mL). Serum concentrations of hepcidin-25 measured by ELISA correlated with hepcidin-25 concentrations measured by using an independent LC-MS assay (r = 0.98, P < 0.001). Hepcidin-25 concentrations were increased in patients with cancer (median 54.8 μg/L, 25%–75% range 23.2–93.5 μg/L, n = 34) and rheumatoid arthritis (median 10.6 μg/L, 25%–75% range 5.9–18.4 μg/L, n = 76) compared with healthy individuals (median 1.20 μg/L, 25%–75% range 0.42–3.07 μg/L, n = 100). CONCLUSIONS The use of 2 monoclonal antibodies in a sandwich ELISA format provides a robust and convenient method for measuring concentrations of the active form of hepcidin. This ELISA should help to improve our understanding of the role of hepcidin in regulating iron metabolism.

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ANURACPREEDA, PANAT, KULLANID TEPSUPORNKUL, and RUNGLAWAN CHAWENGKIRTTIKUL. "Immunodiagnosis of paramphistomosis using monoclonal antibody-based sandwich ELISA for detection ofParamphistomum gracilecirculating 16 kDa antigen." Parasitology 144, no. 7 (February 21, 2017): 899–903. http://dx.doi.org/10.1017/s003118201600264x.

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SUMMARYIn this study, we have produced a monoclonal antibody (MoAb) against 16 kDa antigen ofParamphistomum gracile(16 kDaAgPg), and developed an accurate sandwich enzyme-linked immunosorbent assay (sandwich ELISA) for the detection of circulating 16 kDaAg in the serum and fecal samples from cattle naturally infected withP. gracile. MoAb 1D10 was immobilized on a microtitre plate, and the antigen in the samples was captured and detected with biotinylated rabbit anti-16 kDaAgPg antibody. The lower detection limit of sandwich ELISA was 3·5 pg mL−1, and no cross-reaction with other parasite antigens was evaluated. The reliability of the assay was examined using the serum and fecal samples from cattle naturally infected withP. gracile, Fasciola gigantica, Moniezia benedeni, Trichurissp.,Strongyloidessp., strongylids and non-infected animals. The sandwich ELISA showed the sensitivity, specificity and accuracy at 98·33, 100 and 99·55% (serum samples), and 96·67, 100 and 99·09% (fecal samples). Therefore, this detection method is a rapid and excellent potential assay for the accurate diagnosis of paramphistomosis.

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Himananto, Orawan, Kirana Yoohat, Kannawat Danwisetkanjana, Mallika Kumpoosiri, Sombat Rukpratanporn, Yada Theppawong, Sudtida Phuengwas, et al. "Double antibody pairs sandwich-ELISA (DAPS-ELISA) detects Acidovorax citrulli serotypes with broad coverage." PLOS ONE 15, no. 8 (August 27, 2020): e0237940. http://dx.doi.org/10.1371/journal.pone.0237940.

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Yamashita, J., I. Kobayashi, K. Tatematsu, H. Sezutsu, K. Noda, and H. Ishihara. "Sandwich ELISA Using a Mouse/Human Chimeric CSLEX-1 Antibody." Clinical Chemistry 62, no. 11 (November 1, 2016): 1516–23. http://dx.doi.org/10.1373/clinchem.2016.260968.

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Abstract BACKGROUND An assay using a mouse antisialyl Lewis X (sLeX) antibody (CSLEX-1) is used clinically for screening and monitoring patients with breast cancer in Japan. However, the IgM isoform of CSLEX-1 is not preferred for the assay because the bulkiness of IgM generally causes poor accessibility to the antigen. To solve this problem, we developed an antisLeX mouse/human chimeric IgG antibody, CH-CSLEX-1, using transgenic silkworms. The performance of a homologous sandwich ELISA of CH-CSLEX1 was then evaluated. METHODS To generate CH-CSLEX-1, we used a GAL4/UAS binary gene expression system in transgenic silkworms. The reactivities of CSLEX-1 and CH-CSLEX-1 were determined in a Biacore analysis. To confirm antigen specificity, 3 antigens [sLeX, sLeA, and Lewis Y (LeY)] were used. RESULTS CH-CSLEX-1 formed correctly as an IgG class of immunoglobulin molecule with an isoelectric point close to the predicted value. The best combination for capturing and probing in a sandwich ELISA was determined as a homologous combination of CH-CSLEX-1. The CH-CSLEX-1 assay specifically detected sLeX, but not sLeA and LeY. A correlation analysis with 107 human samples showed good concordance between the conventional CSLEX-1 assay (homologous sandwich ELISA using CSLEX-1) and the CH-CSLEX-1 assay (r = 0.98). Moreover, the CH-CSLEX-1 assay was not affected by either human antimouse IgG antibodies (HAMA IgG) or HAMA IgM. CONCLUSIONS The mouse/human chimeric antibody CH-CSLEX-1 allowed the establishment of a highly specific sandwich ELISA for sLeX that was not affected by HAMA.

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Manolov, V., B. Atanasova, V. Vasilev, K. Tzatchev, and M. Velizarova. "Elisa Method For Serum Hepcidin Quantification In Bulgarian Population." Acta Medica Bulgarica 41, no. 1 (June 1, 2014): 22–29. http://dx.doi.org/10.2478/amb-2014-0003.

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Summary Hepcidin is a 25-aminoacid cysteine-rich iron regulating peptide. Hepcidin quantification in human blood may provide further insights for the pathogenesis of disorders of iron homeostasis and might prove a valuable tool for clinicians for the differential diagnosis of anaemia. This study describes ELISA immunoassay for hepcidin quantification in human serum. We used a sandwich ELISA method from USCN Life Science inc., that consists of ready to use, pre-coated 96-well strip plate with 2 antihepcidin-25 monoclonal antibodies. A recombinant hepcidin in 16 μg/l concentration is used as a standard; it reconstitutes with 1.0 ml standard diluent to prepare a stock solution. We correlated ELISA results of hepcidin-25 measurements in healthy population with hemodialysis patients. The sandwich ELISA was highly specific for hepcidin-25, having a low limit of quantification of 0.020 μg/l. Hepcidin- 25 concentrations were increased in hemodialysis patients (median 33.05 μg/l, range 22.31 -60.98 μg/l, n = 10) compared with healthy individuals (median 12.41 μg/l, range 6.05-18.53 μg/l, n = 40). The use of 2 monoclonal antibodies in a sandwich ELISA format provides a robust, convenient and not very expensive method for measuring concentrations of the active form of hepcidin. It should help to improve our understanding of the role of hepcidin in regulating iron metabolism.

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Tsumuraya, Takeshi, Takeshi Sato, Masahiro Hirama, and Ikuo Fujii. "Highly Sensitive and Practical Fluorescent Sandwich ELISA for Ciguatoxins." Analytical Chemistry 90, no. 12 (May 17, 2018): 7318–24. http://dx.doi.org/10.1021/acs.analchem.8b00519.

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Suzaki, Yuki, Yuri Ozawa, and Hiroyuki Kobori. "Quantification of human angiotensinogen by a novel sandwich ELISA." Peptides 27, no. 11 (November 2006): 3000–3002. http://dx.doi.org/10.1016/j.peptides.2006.05.006.

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Hayashi, Yuzuru, Rieko Matsuda, Tamio Maitani, Katsutoshi Ito, Waka Nishimura, Kazuhiro Imai, and Masako Maeda. "An expression of within-plate uncertainty in sandwich ELISA." Journal of Pharmaceutical and Biomedical Analysis 36, no. 1 (September 2004): 225–29. http://dx.doi.org/10.1016/j.jpba.2004.05.017.

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Vasconcellos, Flávia Aleixo, Mariana Loner Coutinho, Gabriela Hädrich, Leonardo Garcia Monte, Núbia Seyffert, Cláudia Pinho Hartleben Fernandes, and José Antonio Guimarães Aleixo. "Antigen capture sandwich ELISA for the diagnosis of leptospirosis." Veterinary Immunology and Immunopathology 128, no. 1-3 (March 2009): 339. http://dx.doi.org/10.1016/j.vetimm.2008.10.273.

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Han, Huiling, Keith Westerman, Vaughn Ostland, Patrick Gutschow, Gordana Olbina, Jacqueline da Silva Guimarães, Juçara Gastaldi Cominal, et al. "A Novel Dual Monoclonal Sandwich ELISA for Human Erythroferrone." Blood 128, no. 22 (December 2, 2016): 1272. http://dx.doi.org/10.1182/blood.v128.22.1272.1272.

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Abstract Erythroferrone (ERFE) is a hormone produced by erythroblasts in the bone marrow in response to erythropoietin (EPO). Recent animal studies have shown that rather than being involved in regulation of baseline erythropoiesis, ERFE acts as a stress erythropoiesis-specific regulator of hepcidin expression. By suppressing hepcidin expression in the liver, EFRE contributes to increased dietary iron absorption and recycling of stored iron necessary for recovery of blood mass after hemorrhage. In addition, ERFE was found to be involved in hepcidin regulation in inherited iron loading anemias, such as b-Thalassemia. ERFE has potential as a clinical marker for assessing erythropoiesis in patients with blood disorders. To date, there have been no reports of a human ERFE assay development and validation. To study the biological function and potential clinical applications of ERFE in humans, we developed the first dual monoclonal sandwich ELISA for serum measurement. Purified recombinant ERFE was used as antigen to immunize mice and subsequently screen 3000 hybridomas for binding to human ERFE. The nine positive hybridomas were expanded and monoclonal antibody from each clone purified and isotyped. We biotinylated each antibody and queried all possible combinations of capture and detection antibodies for binding activity. We discovered at least two pairs of antibodies suitable for assay optimization. Two mAbs were selected, 4C1 and 2D2, as the capture and detection antibody, respectively. We used streptavidin-HRP to quantify binding and detection of ERFE. The ERFE ELISA standard curve ranges from 0-50 ng/mL. The assay's lower limit of detection (LLOD) is 0.15 ng/mL and lower limit of quantitation (LLOQ) is 0.17 ng/mL. We assessed the normal range of ERFE by analyzing serum from110 healthy first time blood donors with normal iron status determined by assessment of ferritin, plasma iron, and transferrin saturation. Serum ERFE in the first time blood donors ranged from 0.15 to 3.94 ng/mL with a mean of 0.83 ng/mL. To re-capitulate the animal data previously observed in mice (Kautz et al., Nat Genet. 2014; 46(7): 678-684), we examined the effect of blood donation on human serum ERFE concentrations (Figure 1). We analyzed sera from three donors which underwent platelet and plasma-apheresis at baseline and day 2, 3, 4, 5, 7, 9, 10, 14, and 120 (Li et al., J Clin Apher., 2016). It was estimated that 30ml of RBCs were lost in the procedure. Serum ERFE concentrations were elevated from baseline in each patient within 2 days and remained higher through 14 days. At 120 days serum ERFE returned to baseline levels. We went on to test the concept that serum ERFE concentrations would be elevated in blood disorders associated with ineffective erythropoiesis, we obtained serum samples from X-linked sideroblastic anemia probands and 15 of their family members. Nine of the probands had point mutation in the ALAS2 gene and two had a-globin duplications. We measured serum ERFE in the probands and family controls and discovered that ERFE was significantly increased in probands relative to familial controls (Figure 2). Family members had ERFE concentrations similar to healthy first time blood donors (<1 ng/ml). An additional study was conducted to examine ERFE in thalassemia patients whom are known to exhibit ineffective erythropoiesis due to mutations in the α- or β-globin genes that cause production of deformed red blood cells. We obtained sera from patients with both α- and β-thalassemia and discovered that both β+-thalassemia and β0-thalassemia patients had significantly higher serum ERFE concentrations than controls and a group of iron deficient (ID) controls (Figure 3). The β0-thalassemia patients had highly elevated serum ERFE which is likely due to the degree that erythropoiesis is affected. The data we present lends strong support to the quality of the dual monoclonal sandwich ELISA we have developed. The ERFE assay is very sensitive, has excellent reproducibility and spike recovery characteristics, and is easy to perform. The physiological and clinical data we present supports the assertion that the assay is specific for ERFE and will allow insight into a number of hematological diseases. Figure 1 Effect of plasma- or platelet-apheresis on ERFE. Figure 1. Effect of plasma- or platelet-apheresis on ERFE. Figure 2 ERFE in X-linked Sideroblastic Anemia. ****p<0.0001 Figure 2. ERFE in X-linked Sideroblastic Anemia. ****p<0.0001 Figure 3 ERFE in Iron Deficient and Thalassemic Patients. ****p<0.0001, **p<0.005 Figure 3. ERFE in Iron Deficient and Thalassemic Patients. ****p<0.0001, **p<0.005 Disclosures Han: Intrinsic Lifesciences.: Employment, Equity Ownership. Westerman:Intrinsic LifeSciences: Employment. Ostland:Intrinsic LifeScienc s: Employment, Equity Ownership. Gutschow:Intrinsic LifeSciences: Employment, Equity Ownership. Olbina:Intrinsic LifeSciences: Employment, Equity Ownership. Westerman:Intrinsic LifeSciences: Employment, Equity Ownership.

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Abd Elreheim, Amal Mahfouz, Alyaa A. Farid, and Noha A. Mahana. "Sandwich-Elisa Development for the Diagnosis of Toxoplasma Gondii." Journal of the Egyptian Society of Parasitology 46, no. 2 (August 2016): 429–40. http://dx.doi.org/10.12816/0031649.

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Fesus, L., and Gabriella Arato. "Quantitation of tissue transglutaminase by a sandwich ELISA system." Journal of Immunological Methods 94, no. 1-2 (November 1986): 131–36. http://dx.doi.org/10.1016/0022-1759(86)90225-5.

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Shields, J. G., and M. W. Turner. "The importance of antibody quality in sandwich ELISA systems." Journal of Immunological Methods 87, no. 1 (February 1986): 29–33. http://dx.doi.org/10.1016/0022-1759(86)90340-6.

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Granström, M., M. Eriksson, and G. Edevåg. "A sandwich ELISA for bovine serum in viral vaccines." Journal of Biological Standardization 15, no. 3 (January 1987): 193–97. http://dx.doi.org/10.1016/0092-1157(87)90022-9.

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ABD ELREHEIM, AMAL, ALYAA FARID, NOHA MAHANA, IBRAHIUM BAUIOMY, and AZZA ELAMEER. "SANDWICH-ELISA DEVELOPMENT FOR THE DIAGNOSIS OF TOXOPLASMA GONDII." Journal of the Egyptian Society of Parasitology 46, no. 2 (August 1, 2016): 429–40. http://dx.doi.org/10.21608/jesp.2016.88726.

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Todini, Luca, and Alessandro Malfatti. "A leptin sandwich ELISA kit unusable for domestic animals." Chronobiology International 38, no. 8 (June 16, 2021): 1087–88. http://dx.doi.org/10.1080/07420528.2021.1941076.

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de LUIS, R., M. D. PÉREZ, L. SÁNCHEZ, M. LAVILLA, and M. CALVO. "Development of Two Immunoassay Formats To Detect β-Lactoglobulin: Influence of Heat Treatment on β-Lactoglobulin Immunoreactivity and Assay Applicability in Processed Food." Journal of Food Protection 70, no. 7 (July 1, 2007): 1691–97. http://dx.doi.org/10.4315/0362-028x-70.7.1691.

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Milk proteins are commonly used as ingredients in the food industry because of their functional properties, but they can cause severe reactions in milk-allergic individuals. In this work, two enzyme-linked immunosorbent assay (ELISA) formats were developed to detect bovine β-lactoglobulin. The indirect competitive ELISA involved the use of anti–β-lactoglobulin antisera, and the sandwich ELISA involved the use of specific antibodies isolated using a β-lactoglobulin immunosorbent material. The effect of heat treatment on immunoreactivity of the protein in buffer and in milk was determined with both assays. The amount of immunoreactive protein in buffer and in milk decreased as determined by the sandwich ELISA, whereas the amount increased when measuring with the competitive ELISA. Several food products, including meat, bakery products, sauces, and snacks, were analyzed. With both assays, 10 of 11 products in which the ingredient list included the terms “powdered milk” or “milk proteins” contained β-lactoglobulin. However, the β-lactoglobulin concentration in these products obtained with the competitive ELISA were much higher than those obtained with the sandwich ELISA. These differences could be explained by the fact that the determination of β-lactoglobulin concentration by immunoassay is influenced by differences in antibody recognition of the protein present in highly processed foods. Therefore, the antigen-binding properties of antibodies used in a particular immunoassay are important for a correct interpretation of results obtained in food processed at high temperature.

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ANURACPREEDA, PANAT, RUNGLAWAN CHAWENGKIRTTIKUL, and PRASERT SOBHON. "Immunodiagnostic monoclonal antibody-based sandwich ELISA of fasciolosis by detection ofFasciola giganticacirculating fatty acid binding protein." Parasitology 143, no. 11 (June 17, 2016): 1369–81. http://dx.doi.org/10.1017/s0031182016001104.

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SUMMARYUp to now, parasitological diagnosis of fasciolosis is often unreliable and possesses low sensitivity. Hence, the detection of circulating parasite antigens is thought to be a better alternative for diagnosis of fasciolosis, as it reflects the real parasite burden. In the present study, a monoclonal antibody (MoAb) against recombinantFasciola giganticafatty acid binding protein (rFgFABP) has been produced. As well, a reliable sandwich enzyme-linked immunosorbent assay (sandwich ELISA) has been developed for the detection of circulating FABP in the sera of mice experimentally and cattle naturally infected withF. gigantica. MoAb 3A3 and biotinylated rabbit anti-recombinant FABP antibody were selected due to their high reactivities and specificities. The lower detection limit of sandwich ELISA was 5 pg mL−1, and no cross-reaction with other parasite antigens was observed. This assay could detectF. giganticainfection from day 1 post infection. In experimental mice, the sensitivity, specificity and accuracy of this assay were 93·3, 100 and 98·2%, while in natural cattle they were 96·7, 100 and 99·1%. Hence, this sandwich ELISA method showed high efficiencies and precisions for diagnosis of fasciolosis byF. gigantica.

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Nithyanandham Masilamani and Dhanraj Ganapathy. "Awareness about ELISA technique among dental students." International Journal of Research in Pharmaceutical Sciences 11, SPL3 (September 17, 2020): 806–10. http://dx.doi.org/10.26452/ijrps.v11ispl3.3024.

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The enzyme-linked immunosorbent assay (ELISA) can be used to recognize proteins, peptides, antibodies as well as hormones. Often known also as an enzyme immunoassay (EIA), ELISA is used as a diagnostic test in the field of biomedicine and science. This study was conducted to determine the understanding of the ELISA technique among dental students. This survey was performed for assessing the awareness about ELISA technique amongst the dental students. This study was a questionnaire oriented, cross-sectional type of survey comprising 100 dental college students in Chennai. A self-designed questionnaire with 10 questions eliciting the knowledge and awareness about applications of ELISA technique among dental college students. Questionnaires were circulated through an online website survey planet. The questions explored the awareness on ELISA technique diagnostic indications, Direct ELISA, Indirect ELISA, Sandwich ELISA, Competitive ELISA and mechanism of ELISA technique. After the responses were received from 100 participants, data were collected and analysed.67% of the respondents were aware of the ELISA technique .52% were aware of direct ELISA technique. 45%% were aware of the indirect ELISA technique. 42% were aware of the sandwich ELISA technique, 38% were aware of the competitive ELISA technique. 35% were aware of the mechanism of the ELISA technique. The awareness about the ELISA technique in diagnostic medical applications was less among dental students. Increased awareness and educational programs should be initiated to spread knowledge about the ELISA technique among all students and clinicians.

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Veling, J., F. G. van Zijderveld, A. M. van Zijderveld-van Bemmel, H. W. Barkema, and Y. H. Schukken. "Evaluation of Three Newly Developed Enzyme-Linked Immunosorbent Assays and Two Agglutination Tests for Detecting Salmonella enterica subsp. enterica Serovar Dublin Infections in Dairy Cattle." Journal of Clinical Microbiology 38, no. 12 (2000): 4402–7. http://dx.doi.org/10.1128/jcm.38.12.4402-4407.2000.

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In this study test characteristics of three newly developed enzyme-linked immunosorbent assays (ELISAs) for Salmonella enterica subsp. enterica serovar Dublin were evaluated and compared with two agglutination tests. The ELISAs involved were an indirect ELISA with serovar Dublin lipopolysaccharide (LPS ELISA), an indirect ELISA with serovar Dublin flagellar antigen (GP ELISA), and a double-antibody sandwich blocking ELISA that uses monoclonal antibodies against S. enterica subsp.enterica serovar Enteritidis flagellin (GM-DAS ELISA). The agglutination tests involved were two routine serum agglutination tests with either somatic (O) or flagellar (H) antigen. Diagnostic specificity of the three ELISAs was determined using 840 serum samples from seven dairy herds without any history of serovar Dublin infection. Cutoff values at a titer of 100, 100, and 10, respectively, for the LPS ELISA, GP ELISA, and GM-DAS blocking ELISA resulted in a specificity of 99.3, 100, and 100%, respectively. Using these cutoff values the LPS ELISA, GP ELISA, and GM-DAS ELISA were able to detect, respectively, 30, 46, and 38% of 50 fecal culture-positive animals from 13 herds with a recent serovar Dublin infection. With the same cutoff values, active carriers (n = 18) were detected for 94.4% with the LPS ELISA and for 100% with the GP and GM-DAS ELISAs. Kappa values determined on the results of all tests from 8 of the 13 serovar Dublin-infected herds and the 7 control herds demonstrated a good correlation between the results of all ELISAs and the H-agglutination test. The results of the O-agglutination test failed to correlate with those of the other tests. Using a set of sera from 170 aborting cows (with 25 abortions due to serovar Dublin), test results of the ELISAs and the H-agglutination test were comparable. The H-agglutination test may be used successfully for single sample testing, especially to diagnose abortion due to serovar Dublin. It is concluded that the ELISAs are useful diagnostic tools in serovar Dublin control programs and that they are preferred to agglutination tests for reasons of automation and costs.

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Schoenthaler, S. L., and S. Kapil. "Development and Applications of a Bovine Coronavirus Antigen Detection Enzyme-Linked Immunosorbent Assay." Clinical Diagnostic Laboratory Immunology 6, no. 1 (January 1, 1999): 130–32. http://dx.doi.org/10.1128/cdli.6.1.130-132.1999.

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ABSTRACT We developed a monoclonal antibody-based, antigen capture sandwich enzyme-linked immunosorbent assay (ELISA) for bovine coronavirus. We compared the ELISA with electron microscopy and the hemagglutination test and found a close correlation between them. The sensitivity of the ELISA was 104 bovine coronavirus particles per ml of 10% fecal suspension. Compared with electron microscopy, bovine coronavirus ELISA had 96% specificity.

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Himananto, Orawan, Petcharat Thummabenjapone, Plearnpis Luxananil, Mallika Kumpoosiri, Ratchanee Hongprayoon, Wichai Kositratana, and Oraprapai Gajanandana. "Novel and Highly Specific Monoclonal Antibody to Acidovorax citrulli and Development of ELISA-Based Detection in Cucurbit Leaves and Seed." Plant Disease 95, no. 9 (September 2011): 1172–78. http://dx.doi.org/10.1094/pdis-12-10-0889.

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A novel monoclonal antibody (MAb) specific to the seedborne bacterium Acidovorax citrulli was produced. MAb 11E5 reacted specifically with 19 strains of A. citrulli but not with three closely related bacteria in the family Comamonadaceae (i.e., A. facilis, Comamonas acidovorans, and C. testosteroni) and another seven phytopathogenic bacteria. Moreover, this MAb detected a strain of A. citrulli that was not detected by a commercial enzyme-linked immunosorbent assay (ELISA)-based kit and a commercial immunochromatographic strip test. In Western blot analysis, MAb 11E5 reacted with an A. citrulli protein of a molecular mass >170 kDa. MAb 11E5 was employed to develop two sandwich ELISA systems: MAb captured-sandwich ELISA (MC-sELISA) and polyclonal antibody captured-sandwich ELISA (PC-sELISA). MC-sELISA was 10 times more sensitive than PC-sELISA for detection of A. citrulli in cucurbit leaf and seed extracts. The detection limit of the MC-sELISA was 5 × 104 CFU/ml. Detection of A. citrulli in naturally infected cucurbit leaves, fruit, and seed was also feasible using MC-sELISA. The newly established MCsELISA provides another alternative for specific detection of A. citrulli in cucurbits and can be applied for routine field inspection.

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