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Literatura académica sobre el tema "Hybridoma cell"

Autor: Grafiati

Publicado: 10 de mayo de 2025

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Artículos de revistas sobre el tema "Hybridoma cell"

Ozaki, S., S. K. Durum, K. Muegge, J. York-Jolley y J. A. Berzofsky. "Production of T-T hybrids from T cell clones. Direct comparison between cloned T cells and T hybridoma cells derived from them." Journal of Immunology 141, n.º 1 (1 de julio de 1988): 71–78. http://dx.doi.org/10.4049/jimmunol.141.1.71.

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Abstract It has been assumed, without direct evidence, that T cell hybridomas and non-transformed T cell clones are both good models of normal Ag-specific T cells. To compare directly the difference in activation of cloned normal T cells and T hybridoma cells with the same TCR, cloned T hybridoma cells were obtained by fusing pre-established, myoglobin-specific, Iad-restricted T cell clones (14.5 and 9.27) with BW5147 cells. T cell clones were pre-activated with IL-2 as well as specific Ag before fusion. Cloned T hybridoma A3.4C6 was derived from Lys 140-specific and I-Ed-restricted clone 14.5. The other cloned T hybridoma, C7R14, was a fusion product of Glu 109-specific and I-Ad-restricted clone 9.27. Both T hybridomas showed the same Ag specificity and Ia restriction as the parental cloned T cells. However, C7R14 showed higher apparent affinity and broader cross-reactivity than 9.27. Clone 14.5, but not hybridoma A3.4C6, appeared to stimulate splenic cells to secrete cytokines inhibiting HT-2A cell proliferation. The most striking difference between the clones and hybridomas was that both clones, but neither of the matched hybridomas, were induced to synthesize IL-1 on stimulation with Ag. Finally, both cloned T cells and T hybridomas killed Ag-pulsed Iad-bearing B lymphoma target cells. This evidence suggests that killing function can be inherited from clones to hybridomas. However, the clones were much more efficient at killing than the hybridomas, and the hybridomas were more efficient at IL-2 production than the clones. Thus, matched pairs of clones and hybridomas differ in their capacity to mediate the two functions or may tend to be selected differently during cloning. Thus, although our results generally support the validity of T cell hybridomas as faithful models of the corresponding T cell clones, a number of subtle and not-so-subtle differences indicate that caution must be used in such an extrapolation.

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Hawksworth, David, Jeff Moore y Isaac Larkin. "Diversity and affinity from plasma vs. B cells for monoclonal antibody development". Journal of Immunology 212, n.º 1_Supplement (1 de mayo de 2024): 0234_4543. http://dx.doi.org/10.4049/jimmunol.212.supp.0234.4543.

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Abstract Diagnostic assay development requires a diverse panel of high affinity antibodies. Historically, our lab has used splenic CD19+ B-cells to generate mouse hybridoma cell lines. However, literature suggests that, as a group, terminally differentiated, CD138+ plasma cells secrete the highest affinity antibodies, making these cells an attractive target for hybridoma development. To compare the use of these two lymphocyte populations for generating mouse hybridomas, three fusion experiments were completed whereby cells were separately isolated and fused. Hybridomas were plated in semi-solid media, expanded, and selected based on size and antibody secretion. An average of 246 (+ 73) plasma cell hybridoma colonies were available for selection, per 1x106 specific lymphocytes used for fusion, compared to 19 (+ 13) B cell hybridoma colonies. Cells from selected colonies were transferred to 96-well plates, grown for 7 days and the antibodies screened for antigen binding by enzyme immunoassay. Across the three fusion experiments, 40 (+19)% of the wells with growth from the plasma cell hybridomas were antigen reactive, vs. 6 (+ 0.5)% from the B cell hybridomas. Our data suggests a higher fusion efficiency from the plasma cell population, which allows for a greater sampling of the mouse immune repertoire, leading to identification of more antigen specific antibodies with a wider range of affinities and greater sequence diversity.

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Iwata, M., K. Katamura, R. T. Kubo y K. Ishizaka. "Relationship between T cell receptors and antigen-binding factors. I. Specificity of functional T cell receptors on mouse T cell hybridomas that produce antigen-binding T cell factors." Journal of Immunology 143, n.º 12 (15 de diciembre de 1989): 3909–16. http://dx.doi.org/10.4049/jimmunol.143.12.3909.

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Abstract Upon antigenic stimulation with OVA-pulsed syngeneic macrophages, the mouse T cell hybridoma 231F1 produced glycosylation inhibiting factor (GIF) having affinity for OVA and IgE-suppressive factors, whereas another T cell hybridoma, 12H5, cells produced OVA-binding glycosylation enhancing factor (GEF) and IgE-potentiating factor. The OVA-binding GIF from the 231F1 cells is an Ag-specific Ts cell factor, whereas OVA-binding GEF from the 12H5 cells is an Ag-specific augmenting factor. Both hybridomas express CD3 complex and functional TCR-alpha beta. Cross-linking of TCR-alpha beta or CD3 molecules on the hybridomas by anti-TCR-alpha beta mAb or anti-CD3 mAb and protein A resulted in the formation of the same factors as those obtained by the stimulation of the cells with OVA-pulsed syngeneic macrophages. It was also found that both the 231F1 cells and 12H5 cells formed IgE-binding factors upon incubation with H-2d and H-2b APC, respectively, with a synthetic peptide corresponding to residues 307-317 in the OVA molecules (P307-317). Six other synthetic peptides, including those containing the major immunogenic epitope, i.e., P323-339, failed to stimulate the hybridomas in the presence of APC. Indeed, all of the 10 T cell hybridoma clones, which could produce either OVA-binding GIF or OVA-binding GEF, responded to P307-317 and APC for the formation of IgE-binding factors. In contrast, GIF/GEF derived from six other hybridoma clones, whose TCR recognized P323-339 in the context of a MHC product, failed to bind to OVA-coupled Sepharose. The results indicate the correlation between the fine specificity of TCR and the affinity of GIF/GEF to the nominal Ag. The amino acid sequence of P307-317 suggested that TCR on the cell sources of Ag-binding factors are specific for an external structure of the Ag molecules.

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Tiebout, R. F., F. van Boxtel-Oosterhof, E. A. Stricker y W. P. Zeijlemaker. "A human hybrid hybridoma." Journal of Immunology 139, n.º 10 (15 de noviembre de 1987): 3402–5. http://dx.doi.org/10.4049/jimmunol.139.10.3402.

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Abstract Hybrid hybridomas are obtained by fusion of two cells, each producing its own antibody. Several authors have reported the construction of murine hybrid hybridomas with the aim to obtain bispecific monoclonal antibodies. We have investigated, in a model system, the feasibility of constructing a human hybrid hybridoma. We fused two monoclonal cell lines: an ouabain-sensitive and azaserine/hypoxanthine-resistant Epstein-Barr virus-transformed human cell line that produces an IgG1 kappa antibody directed against tetanus toxoid and an azaserine/hypoxanthine-sensitive and ouabain-resistant human-mouse xenohybrid cell line that produces a human IgG1 lambda antibody directed against hepatitis-B surface antigen. Hybrid hybridoma cells were selected in culture medium containing azaserine/hypoxanthine and ouabain. The hybrid nature of the secreted antibodies was analyzed by means of two antigen-specific immunoassays. Our results show that it is possible, with the combined use of transformation and xenohybridization techniques, to construct human hybrid hybridomas that produce bispecific antibodies.

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Iwata, M., M. Adachi y K. Ishizaka. "Antigen-specific T cells that form IgE-potentiating factor, IgG-potentiating factor, and antigen-specific glycosylation-enhancing factor on antigenic stimulation." Journal of Immunology 140, n.º 8 (15 de abril de 1988): 2534–42. http://dx.doi.org/10.4049/jimmunol.140.8.2534.

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Abstract BDF1 mice were immunized with alum-absorbed OVA and T cell hybridomas were constructed from their splenic T cells. Many of the hybridomas constitutively produced glycosylation enhancing factor (GEF), which could switch the T cell hybridoma 23A4 cells from the formation of IgE-suppressive factors to the formation of IgE-potentiating factors. When one of the hybridoma clones, 12H5, was incubated with OVA-pulsed syngeneic or semi-syngeneic (H-2b) macrophages, the hybridoma produced GEF that have affinity for OVA, but not for either keyhole limpet hemocyanin or BSA. However, the same hybridoma constitutively produced nonspecific GEF, that lacked affinity for OVA. Upon incubation with OVA-pulsed macrophages, the same hybridoma produced both IgE-potentiating factors and IgG-potentiating factors which selectively enhance the IgE response and IgG response, respectively. Both Ag-specific GEF and nonspecific GEF from the hybridoma bind to p-aminobenzamidine-agarose, and are recovered by elution with benzamidine. It was also found that both OVA-specific GEF and nonspecific GEF from the hybridoma induced the release of arachidonic acid from phospholipids of mouse fibrosarcoma cell line, HSDM1C1 cells. GEF formed by the 12H5 hybridoma bound to alloantibodies reactive to the product(s) of the I-Ab subregion of major histocompatibility complex. The Ag-specific GEF consisted of two Mr species, of 70 to 90 kDa and 50 to 60 kDa, whereas nonspecific GEF consisted of 50 to 60 kDa and 25 to 30 kDa molecules. Reduction and alkylation treatment of the OVA-specific GEF resulted in the formation of nonspecific GEF, suggesting that Ag-specific GEF is composed of Ag-binding polypeptide chain and nonspecific GEF.

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Kawasaki, H., C. A. Martin, T. Uchida, M. Usui, T. Noma, M. Minami y M. E. Dorf. "Functional analysis of cloned macrophage hybridomas. V. Induction of suppressor T cell responses." Journal of Immunology 137, n.º 7 (1 de octubre de 1986): 2145–51. http://dx.doi.org/10.4049/jimmunol.137.7.2145.

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Abstract It has been suggested that macrophage-like accessory cells are involved in suppressor T cell (Ts) induction. To further analyze this issue, we obtained several cloned macrophage hybridoma cell lines by somatic cell fusion of the macrophage tumor P388D1 of DBA/2 (H-2d) origin with splenic adherent cells of CKB mice (H-2k). Several cloned lines displayed the serological and functional characteristics of macrophages. We evaluated the ability of these hybridomas to induce third order or effector Ts (Ts3) to suppress the contact sensitivity response against the hapten 4-hydroxy-3-nitrophenyl acetyl (NP). In contrast to the parental P388D1 and two other macrophage hybridomas, one macrophage hybridoma clone, termed 63, when conjugated with NP, induced Ts3, which suppressed contact sensitivity responses against NP but not DNFB, showing that the Ts3 were antigen specific. Macrophage hybridoma 63 could specifically induce Ts3 activity in either H-2k, H-2d, or H-2k/H-2d heterozygous hosts. Thus, macrophage hybridoma 63 functionally expressed major histocompatibility complex-related restricting determinants, and the fusion with cells from a H-2k macrophage donor caused a functional complementation of H-2d-related, Ts-inducing elements. The genetic restriction governing induction of Ts3 was controlled by genes that mapped to I-J region. Furthermore, NP-conjugated macrophage hybridoma 63 could serve as a target for elicitation of suppressor responses after administration of I-Jk, but not I-Jb, restricted suppressor factor. The data suggest that macrophage hybridomas represent a means to dissect heterogeneity within the macrophage population. The data also imply that the I-J determinants expressed on macrophages represent a ligand for the antigen receptor of Ts.

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Lanier, L. L., J. J. Ruitenberg y J. H. Phillips. "Functional and biochemical analysis of CD16 antigen on natural killer cells and granulocytes." Journal of Immunology 141, n.º 10 (15 de noviembre de 1988): 3478–85. http://dx.doi.org/10.4049/jimmunol.141.10.3478.

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Abstract CD16 Ag is associated with the low affinity FcR for IgG expressed on human NK cells and granulocytes. In this study, we demonstrate that NK cells specifically lyse murine anti-CD16 hybridoma cell lines, but do not lyse hybridomas against other cell surface differentiation Ag expressed on NK cells. Moreover, the CD18 structure is involved in the CD16-specific xenogeneic interaction between human effector cells and murine hybridoma target cells. Although interaction with anti-CD16 hybridomas or antibodies triggers the cytolytic mechanism of NK cells, this interaction does not induce cellular proliferation. In contrast to NK cells, CD16+ granulocytes do not lyse anti-CD16 hybridoma cell targets and do not mediate ADCC against antibody-coated human tumor cell targets. These findings indicate a fundamental difference in the antibody-dependent cellular cytotoxicity mechanisms of NK cells and granulocytes. Comparative biochemical analysis of CD16 on NK cells and granulocytes revealed significant differences in the size of the polypeptides obtained after removal of N-linked carbohydrate residues with endo-F and N-glycanase digestion.

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Kuchroo, V. K., M. C. Byrne, E. Greenfield, M. J. Whitters, E. A. Nalefsky, A. Rao, M. Collins y M. E. Dorf. "Transfection of TCR alpha-chains into suppressor and T helper cell hybridomas. Production of suppressor factors with predicted antigen specificity." Journal of Immunology 154, n.º 10 (15 de mayo de 1995): 5030–38. http://dx.doi.org/10.4049/jimmunol.154.10.5030.

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Abstract Conditioned medium from Ag-specific suppressor T cell hybridomas contains soluble factors (TsF) that modulate immune responses in an Ag-specific manner. We previously generated a series of TCR-alpha- and TCR-beta- expression variants from a 4-hydroxy-3-nitrophenyl acetyl (NP)-specific inducer suppressor T cell hybridoma and demonstrated that loss of TCR alpha-chain mRNA, but not TCR-beta chain mRNA, was accompanied by concomitant loss of suppressor bioactivity. Suppressor factor bioactivity was restored by expression of TCR alpha-chain cDNA, suggesting that the TCR alpha-chain plays a critical role in Ag-specific suppressor cell function. We have now transfected TCR alpha-chain from a Th cell clone specific for arsanylated peptides plus I-Ad into a TCR-alpha- derivative of an NP-specific inducer suppressor T cell hybridoma. The transfectants expressed a new hybrid TCR-alpha beta complex and produced soluble factors that suppressed azobenzenearsonate hapten (ABA) but not NP delayed-type hypersensitivity responses. These supernatants mediated suppression of the induction, but not the effector phase of the delayed-type hypersensitivity reaction. In reciprocal experiments we transfected a TCR alpha-chain from an NP-specific suppressor T cell hybridoma into a TCR-alpha- hybridoma derived from the ABA-specific Th cell hybridoma. The NP-specific TCR alpha-chain was expressed in the Th cell hybridoma, but the supernatant from this transfectant did not suppress DTH responses to either NP or ABA. However, the latter supernatants, when combined with cell lysates derived from a TCR-alpha- Ts hybridoma, specifically suppress NP DTH responses. These data are consistent with the interpretation that TCR alpha-chain imparts Ag specificity to the suppressor molecule and a second, yet undefined, component produced by the Ts hybridoma controls the immunoregulatory bioactivity.

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Takahashi, M. y S. A. Fuller. "Production of murine hybrid-hybridomas secreting bispecific monoclonal antibodies for use in urease-based immunoassays." Clinical Chemistry 34, n.º 9 (1 de septiembre de 1988): 1693–96. http://dx.doi.org/10.1093/clinchem/34.9.1890.

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Abstract To produce bispecific antibodies (BiAbs) for enzyme immunoassay (EIA) to replace antibody-enzyme conjugates, we developed a panel of 8-azaguanine/ouabain-resistant anti-urease variant hybridoma cell lines for use in hybridoma-hybridoma fusions. These variants represent mouse immunoglobulin subclasses IgM, IgG1, IgG2a, and IgG2b and have growth rates equal to those of the parental hybridomas. We fused an anti-urease-secreting variant hybridoma with an anti-human choriogonadotropin (hCG)-secreting hybridoma (both of IgG1 subclass) and selected the desired product with growth media containing hypoxanthine-aminopterin-thymidine (HAT) and ouabain. Over 95% of the resulting hybrids secreted anti-urease, and 60% of these secreted anti-hCG. The bispecific nature of secreted antibodies was demonstrated in a simultaneous EIA where BiAbs, hCG, and urease (EC 3.5.1.5) were incubated together in anti-hCG-coated microwells. As little as 25 int. units of hCG per liter could be reliably detected, which is equivalent to that for antibody-enzyme conjugates in EIA.

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Dzhambazov, Balik, Tsvetelina Batsalova, Patrick Merky, Franziska Lange y Rikard Holmdahl. "NIH/3T3 Fibroblasts Selectively Activate T Cells Specific for Posttranslationally Modified Collagen Type II". International Journal of Molecular Sciences 24, n.º 13 (28 de junio de 2023): 10811. http://dx.doi.org/10.3390/ijms241310811.

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It has been shown that synovial fibroblasts (SF) play a key role in the initiation of inflammation and joint destruction, leading to arthritis progression. Fibroblasts may express major histocompatibility complex class II region (MHCII) molecules, and thus, they could be able to process and present antigens to immunocompetent cells. Here we examine whether different types of fibroblasts (synovial, dermal, and thymic murine fibroblasts, destructive LS48 fibroblasts, and noninvasive NIH/3T3 fibroblasts) may be involved in the initiation of rheumatoid arthritis (RA) pathogenesis and can process and present type II collagen (COL2)—an autoantigen associated with RA. Using a panel of MHCII/Aq-restricted T-cell hybridoma lines that specifically recognize an immunodominant COL2 epitope (COL2259–273), we found that NIH/3T3 fibroblasts activate several T-cell clones that recognize the posttranslationally glycosylated or hydroxylated COL2259–273 epitope. The HCQ.3 hybridoma, which is specific for the glycosylated immunodominant COL2 epitope 259–273 (Gal264), showed the strongest response. Interestingly, NIH/3T3 cells, but not destructive LS48 fibroblasts, synovial, dermal, or thymic fibroblasts, were able to stimulate the HCQ.3 hybridoma and other COL2-specific T-cell hybridomas. Our experiments revealed that NIH/3T3 fibroblasts are able to activate COL2-specific T-cell hybridomas even in the absence of COL2 or a posttranslationally modified COL2 peptide. The mechanism of this unusual activation is contact-dependent and involves the T-cell receptor (TCR) complex.

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Tesis sobre el tema "Hybridoma cell"

Damico, Nicole. "Preparing and Cloning a Natural Killer Cell Hybridoma". Youngstown State University / OhioLINK, 2000. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1004458025.

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Wilson, James Samuel. "Process intensification of hybridoma cell fermentation". Thesis, University of Edinburgh, 1992. http://hdl.handle.net/1842/12155.

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Monoclonal antibodies can be produced in culture fluid by the fermentation of specificially selected hybridoma cells. Hybridoma cells exhibit suspension type fermentation characteristics and therefore the simplest method for large scale fermentation is that of the stirred tank fermenter. However, such is the growing demand for monoclonal antibodies, methods for increasing the production capacity of a commercial process are being developed. This study examines some of the current process intensification methods in relation to an established production facility. As well as examining the actual productivity increases possible with any method, the applicability of that method to a commercial environment is taken into account. Hollow-fibre systems are investigated, with a potential increase in productivity which was outweighed by the significant retooling and retraining costs. Gel Bead entrapment systems are shown to have great promise, as they can be readily placed into existing equipment and production methods. However, all methods examined, including alginate bead entrapment, were found unsuitable for hybridoma cell culture. A novel method for cell entrapment was developed, using an agarose/alginate gel mixture which allowed greatly improved growth and consistent antibody production. The entrapment method was examined in a continuous chemostatic system. This system was then scaled-up and applied to the existing facility, to give a 25L airlift operating in a chemostatic mode at a rate of 1.2-1.5 day^-1.

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Amribt, Zakaria. "Macroscopic modelling of hybridoma cell fed-batch cultures with overflow metabolism: model-based optimization and state estimation". Doctoral thesis, Universite Libre de Bruxelles, 2014. http://hdl.handle.net/2013/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/209279.

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Monoclonal antibodies (MAbs) have an expanding market for use in diagnostic and therapeutic applications. Industrial production of these biopharmaceuticals is usually achieved based on fed-batch cultures of mammalian cells in bioreactors (Chinese hamster ovary (CHO) and Hybridoma cells), which can express different kinds of recombinant proteins. In order to reach high cell densities in these bioreactors, it is necessary to carry out an optimization of their production processes. Hence, macroscopic model equations must be developed to describe cell growth, nutrient consumption and product generation. These models will be very useful for designing the bioprocess, for developing robust controllers and for optimizing its productivity.

This thesis presents a new kinetic model of hybridoma cell metabolism in fed batch culture and typical illustration of a systematic methodology for mathematical modelling, parameter estimation and model-based optimization and state estimation of bioprocesses.

In the first part, a macroscopic model that takes into account phenomena of overflow metabolism within glycolysis and glutaminolysis is proposed to simulate hybridoma HB-58 cell cultures. The model of central carbon metabolism is reduced to a set of macroscopic reactions. The macroscopic model describes three metabolism states: respiratory metabolism, overflow metabolism and critical metabolism. The model parameters and confidence intervals are obtained via a nonlinear least squares identification. It is validated with experimental data of fed-batch hybridoma cultures and successfully predicts the dynamics of cell growth and death, substrate consumption (glutamine and glucose) and metabolites production (lactate and ammonia). Based on a sensitivity analysis of the model outputs with respect to the parameters, a model reduction is proposed.

In the next step, the effort is directed to the maximization of biomass productivity in fed-batch cultures of hybridoma cells based on the overflow metabolism model. Optimal feeding rate, on the one hand, for a single feed stream containing both glucose and glutamine and, on the other hand, for two separate feed streams of glucose and glutamine are determined using a Nelder-Mead simplex optimization algorithm. Two different objective functions (performance criteria) are considered for optimization; the first criterion to be maximized is the biomass productivity obtained at the end of the fed-batch culture, the second criterion to be minimized is the difference between global substrate consumption and the maximum respiratory capacity.

The optimal multi exponential feed rate trajectory improves the biomass productivity by 10% as compared to the optimal single exponential feed rate. Moreover, this result is validated by the one obtained with the analytical approach in which glucose and glutamine are fed to the culture so as to control the hybridoma cells at the critical metabolism state, which allows maximizing the biomass productivity. The robustness analysis of optimal feeding profiles obtained with different optimization strategies is considered, first, with respect to parameter uncertainties and, finally, with respect to model structure errors.

Finally, the overflow metabolism model is used to develop an extended Kalman filter for online estimation of glucose and glutamine in hybridoma cell fed-batch cultures based on the considered available measurements (biomasses (on-line), lactate and ammonia (on-line or off-line)). The observability conditions are examined, and the performances are analysed with simulations of hybridoma cell fed-batch cultures. Glutamine estimation sensitivity is enforced by minimizing a cost function combining a usual least-squares criterion with a state estimation sensitivity criterion.

Doctorat en Sciences de l'ingénieur
info:eu-repo/semantics/nonPublished

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Faraday, David Brian Foster. "The mathematical modelling of the cell cycle of a hybridoma cell line". Thesis, University of Surrey, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.341620.

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Costello, Mark Eugene. "Growth and productivity of hybridoma cell lines in vitro". Thesis, Manchester Metropolitan University, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.280629.

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El, Debs Bachir. "Functional single-cell hybridoma screening using droplet-based microfluidics". Strasbourg, 2011. http://www.theses.fr/2011STRA6182.

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µm de diamètre) pour la culture de cellules mammifères. On a utilisé ce système pour sélectionner spécifiquement des cellules hybridomes sécrétant des anticorps inhibant l’Enzyme de Conversion de l’Angiotensine. -1 (ECA-1). L’émulsion contenant les cellules encapsulées dans les gouttes a été incubé pendant 6 heurres pour obtenir une quantité considérable d’anticorps avec l’ECA-1. Ensuite, cette émulsion a été réinjectée dans une puce microfluidique, fusionnée avec des gouttes contenant un mélange réactionnel permettant l’obtention d’un signal fluorescent d° à l’activité de l’ECA-1. Les gouttes ayant une faible intensité de fluorescence ont été triées. Une variance considérable dans le taux d’expression d’anticorps a été constatée au niveau mono-cellulaire au sein d’une même lignée de cellules hybridomes o_ les cellules exprimant un taux élevé d’anticorps ont été isolées et cultivées. Ce système permet le criblage de 5_104 cellules par heure et pourra être utiliser pour le criblage de lymphocytes B non immortalisées
This thesis describes a microfluidic platform allowing the functional screening of hybridoma cells on the single-cell level. In this system, individual cells from a heterogeneous population are encapsulated into aqueous microdroplets of a water-in-oil emulsion and assayed directly for the release of antibodies inhibiting drug targets. The microfluidic setup comprises a novel fully integrated chip which allows reinjection, fusion and sorting of droplets sufficiently large (~100 µm in diameter) for the cultivation of mammalian cells. We successfully used this device for the specific selection of hybridoma cells releasing antibodies inhibiting angiotensin converting enzyme-1 (ACE-1). After cell encapsulation, the resulting emulsion was incubated off-chip for 6h to obtain significant antibody concentrations. Subsequently, the droplets were reinjected into another chip, fused with a second droplet species containing all components of a fluorescence assay for ACE-1 activity, and droplets with low fluorescence intensity (indicating ACE-1 inhibition) were sorted. A wide variance in antibody expression levels at the single-cell level within a single hybridoma line was observed and high expressors could be sorted and recultivated. The approach enabled screening more than 5_104 cells per hour and should even be applicable to non-immortalized primary B-cells, as no cell proliferation is required

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Okerlund, Linda Susan. "Energy consumption among static and proliferating hybridoma cell populations". Diss., The University of Arizona, 1991. http://hdl.handle.net/10150/185447.

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To investigate the effects of proliferation on metabolism and cell product yields, proliferating and growth-limited EPOBF7 hybridoma cells have been compared as to their growth rates, energy demand, relative energy distributions, and monoclonal antibody (MAb) yield. Medium deprivation of leucine or serum was used to prevent growth. Energy consumption rates were determined in cell suspensions from rates of glucose consumption, lactate production, and oxygen utilization. In addition, the energy consumption of pathways critical to cell growth and survival were estimated from the relative decreases occurring in oxygen and glucose consumption upon pathway inhibition. The overall rate of energy consumption was significantly lower among growth-limited cultures. In addition, the distributions of oxidative and glycolytic energy among cellular synthetic pathways differed significantly between the culture conditions. Non-growing cultures also produced significantly lower antibody yields. Cell growth was also investigated using ³¹p nuclear magnetic resonance (NMR) spectroscopy of cells grown and maintained in bioreactor culture. Saturation transfer methods detected measurable transfer between inorganic phosphate (P(i)) and the gamma resonance of ATP. This transfer rate could be correlated with cellular growth rates within the reactor. Transfer of magnetization from the gamma resonance of ATP to P(i) was also detected, although the rate of this transfer did not appear to be related to the growth rate. The ratio of these transfer rates was consistently near 4. This information is believed to suggest the importance of other reactions through which ATP may donate its terminal phosphate. Cells in bioreactor culture were found to grow more slowly and produce lower levels of monoclonal antibody when compared to cells proliferating in suspension. such phenomena may be a function of diffusion limitations within the reactor such that the cells cannot obtain the nutrient supply required for optimal cell growth or product formation.

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Futch, William S. Jr. "Dissection of Microphage Activation Using T Cell Hybridoma Derived Lymphokines". VCU Scholars Compass, 1985. http://scholarscompass.vcu.edu/etd/4566.

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Macrophage phenotype/function can be modulated by various T-cell lymphokines (LK). The alteration of macrophage phenotype is a result of LK concentration, duration of exposure, and the level of macrophage activation when obtained from in vivo sources through elicitation by either sterile irritants or immune cellular mechanisms. In order to dissect macrophage activation into discrete signals T cell hybridomas were constructed by fusing HAT sensitive BW5147 cells with nylon-wool purified, con A stimulated T cells. The resulting T cell hybrids were screened for their ability to: (a) protect macrophages from the cytopathic effect of Naegleria; (b) induce class II MHC gene product (Ia antigen) expression; (c) increase cytostasis and tumoricidal activity; and (d) alter ectoenzyme profiles on either resident or thioglycollate (TG) elicited macrophages. Two hybridomas (T-3 and T-9) were selected for further evaluation because of their activity patterns. Supernatants from T-3 and T-9 were compared with cloned Y-interferon (γ-IFN) for alteration of biological activities. Both T-3 and T-9 were able to protect resident macrophage cells from Naegleria but had no protective effect on TG-macrophages. T-9 supernatant had patterns of activity similar to γ-IFN while T-3 patterns were different. The addition of anti-Y-IFN removed T-9 cytostatic activity while not affecting T-3 induced activity. The LK inducing protection from the cytopathic effect of Naegleria lysate is not γ-IFN but another molecular moiety. It was also shown that γ-IFN does not protect TG-macrophages from the destructive effects of adenylate cyclase produced by Bordetella pertussis. We conclude that activation of macrophages for the destruction of tumor cells and activation for protection against amoeba and bacteria occur via different biological pathways. Furthermore, we have proposed an association between the cell cycle and the responsiveness of resident and TG-elicited macrophages to specific LK.

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Balcarcel, R. Robert. "Effects of rapamycin and insulin on the cell cycle and apoptosis of hybridoma cell cultures". Thesis, Massachusetts Institute of Technology, 1999. http://hdl.handle.net/1721.1/85361.

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Rousseau, Fanny. "Systèmes microfluidiques pour la génération d'hybridomes et d'anticorps monoclonaux". Electronic Thesis or Diss., université Paris-Saclay, 2025. http://www.theses.fr/2025UPASQ013.

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Les anticorps sont des molécules produites par le système immunitaire, caractérisées par une grande spécificité de liaison pour un antigène donné, ce qui en fait des outils biologiques puissants pour des applications thérapeutiques et de diagnostic. La production en quantité d'anticorps monoclonaux in vitro est rendue possible en 1975 grâce à la technique des hybridomes. Bien que cette technique soit simple, facile à implémenter et peu coûteuse, ses faibles rendements limitent aujourd'hui son utilisation et ont donné lieu à des méthodes plus modernes de production d'anticorps, qui présentent toutefois de nouvelles limitations.Dans ce contexte, l'objectif de cette thèse est d'améliorer la technique des hybridomes actuelle afin de la repositionner comme une technologie innovante et attractive. En particulier, il s'agit d'implémenter trois dispositifs microfluidiques, intervenant aux différentes étapes du processus de production d'anticorps monoclonaux, pour en optimiser les rendements et en faciliter la procédure.La première partie de ce projet est consacrée à l'identification et à l'isolement des cellules sécrétrices d'anticorps issues de rates de souris immunisées. Après fusion avec les cellules de myélome, ces cellules favorisent la génération d'hybridomes sécrétant des anticorps spécifiques de l'antigène cible. Le but est ainsi de pouvoir réaliser des fusions cellulaires ciblées pour limiter la génération d'hybridomes non fonctionnels. Pour cela, après identification par cytométrie en flux à l'aide d'un panel de marqueurs membranaires ad hoc, l'isolement des cellules d'intérêt est réalisé à l'aide d'un tri magnétique intégré en puce microfluidique.La deuxième partie de la thèse porte sur le développement d'une puce microfluidique dédiée à la fusion cellulaire par voie chimique, en utilisant le polyéthylène glycol (PEG). Ce dispositif vise à optimiser les conditions de fusion entre des cellules de rate et de myélome, et à améliorer les rendements de la méthode conventionnelle. Une version alternative de cette puce, adaptée à la fusion cellulaire par électroporation est également démontrée.Enfin, la dernière partie de cette thèse est consacrée à la sélection des hybridomes sécréteurs d'anticorps spécifiques de la cible d'intérêt par criblage en microfluidique de gouttes. Cette partie, réalisée en collaboration avec l'entreprise strasbourgeoise MicroOmix, permet de simplifier et d'accélérer les étapes post fusion de sélection des clones d'intérêt employées dans la technique des hybridomes
Antibodies are molecules produced by the immune system and are characterized by their high binding affinity and specificity for a given antigen, thus making them powerful biological tools for therapeutic and diagnostic applications. The in vitro production of antibodies was made possible in 1975 by the development of the hybridoma technology. This technique is simple, easy to implement and inexpensive, but its use has been limited by low yields, which has led to the emergence of more modern methods that present their own set of challenges.The central aim of this thesis is to unlock the existing hybridoma technique, repositioning it as an efficient and appealing technology. In particular, the objective is to implement three microfluidic devices at each step of the monoclonal antibody production process in order to optimise yields and facilitate the procedure.The first part of this project is focused on the identification and isolation of antibody-secreting cells from the spleen of immunized mice. Following fusion with myeloma cells, this cell subset may facilitate the generation of hybridomas with the ability to secret antibodies that are specific to the target antigen. Our aim is thus to perform targeted fusions between myeloma and antibody-secreting cells, thereby preventing the generation of non-functional hybridomas. To achieve this objective, the cells of interest are identified by flow cytometry using a dedicated surface markers panel. These cells subset are then isolated using an integrated microfluidic magnetic cell-sorting chip.The second part of the thesis concerns the development of a microfluidic chip dedicated to chemical cell fusion, using polyethylene glycol (PEG). The objective of this device is to optimize the conditions for efficient fusion between spleen and myeloma cells, and to improve the yields of the conventional method in tube. An alternative version of this chip, adapted for cell fusion by electroporation, is also demonstrated.Finally, the last part of this project illustrates the potential of droplet-based microfluidics for the single-cell selection and high-throughput screening of hybridomas that secret antigen-specific antibodies. This demonstration, carried out in collaboration with the Strasbourg-based company MicroOmix, aims to simplify and accelerate the post-fusion steps of the hybridoma technology

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Libros sobre el tema "Hybridoma cell"

Murray, Kevin. Metabolic management of a hybridoma cell line. Manchester: University of Manchester, 1994.

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Welsh, Jonathan Peter. Relationships between hybridoma cell mechanical properties and physiology. Birmingham: University of Birmingham, 1998.

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Guérin, Paul. Role of oxidative stress in the induction of cell death in the hybridoma cell line SP2/0-Ag14. Sudbury, Ont: Laurentian University, School of Graduate Studies, 2004.

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1946-, Taussig Michael J., ed. T cell hybridomas. Boca Raton, Fla: CRC Press, 1985.

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J, Taussig Michael, ed. T cell hybridomas. Boca Raton, Fla: CRC Press, 1985.

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LeFort, Natalie. Characterisation of NAD metabolism and evaluation of the role of NAD-consuming enzymes in apoptosis of Sp2/0-Ag14 murine hybridoma cell line. Sudbury, Ont: Laurentian University, 2004.

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Robert, Hay, ed. Catalogue of cell lines and hybridomas. 6^a ed. Rockville, Md: American Type Culture Collection, 1988.

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Karns, Tanya. The behaviour of hybridoma cells in culture. Sudbury, Ont: Laurentian University, 2003.

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Koski, Pamela. The regulation of GADD 153 in hybridoma cells. Sudbury, Ont: Laurentian University, 2003.

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Oancea, Adriana Ecaterina. Immunoglobulin heavy chain gene expression in hybridoma cells. Ottawa: National Library of Canada = Bibliothèque nationale du Canada, 1997.

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Capítulos de libros sobre el tema "Hybridoma cell"

McDonnell, Susan. "Production of Antibodies in Hybridoma and Non-hybridoma Cell Lines". En Cell Engineering, 65–88. Cham: Springer International Publishing, 2014. http://dx.doi.org/10.1007/978-3-319-10320-4_3.

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Klebe, Robert J. y Kevin L. Bentley. "Chemically Mediated Cell Fusion". En Methods of Hybridoma Formation, 77–96. Totowa, NJ: Humana Press, 1987. http://dx.doi.org/10.1007/978-1-4612-4826-2_4.

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Westerwoudt, Regine J. J. M. "Proliferation and Immune Secretion of B-Cell Hybridomas". En Methods of Hybridoma Formation, 209–30. Totowa, NJ: Humana Press, 1987. http://dx.doi.org/10.1007/978-1-4612-4826-2_10.

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Conrad, Mary K. y Mathew M. S. Lo. "B-Cell Hybridoma Production by Avidin-Biotin Mediated Electrofusion". En Electromanipulation in Hybridoma Technology, 89–102. London: Palgrave Macmillan UK, 1989. http://dx.doi.org/10.1007/978-1-349-11339-2_5.

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Rock, Kenneth L. "Functional T-Cell Hybridomas". En Hybridoma Technology in the Biosciences and Medicine, 527–44. Boston, MA: Springer US, 1985. http://dx.doi.org/10.1007/978-1-4684-4964-8_33.

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Rodrigues, Maria Teresa A., Isaias Raw y Ana Maria Moro. "Residual DNA from Hybridoma Cultures: Decorrence of Apoptosis?" En Animal Cell Technology, 467–72. Dordrecht: Springer Netherlands, 1997. http://dx.doi.org/10.1007/978-94-011-5404-8_75.

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Shay, Jerry W. "Mechanisms of Cell Fusion and Selection in the Generation of Hybridomas". En Methods of Hybridoma Formation, 63–75. Totowa, NJ: Humana Press, 1987. http://dx.doi.org/10.1007/978-1-4612-4826-2_3.

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Romein, B., I. Q. I. O. Melchy, C. Hellinga, J. P. Van Dijken y K. Ch A. M. Luyben. "Monitoring and Modelling Hybridoma Cultures". En Animal Cell Technology: Developments Towards the 21st Century, 847–50. Dordrecht: Springer Netherlands, 1995. http://dx.doi.org/10.1007/978-94-011-0437-1_135.

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Franěk, F. y K. Šrámková. "Unbalanced Media for Hybridoma Cell Culture —an Alternative Reality". En Animal Cell Technology, 675–80. Dordrecht: Springer Netherlands, 1997. http://dx.doi.org/10.1007/978-94-011-5404-8_106.

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Franěk, F. y T. Vomastek. "Spontaneous Apoptosis in Mouse Hybridoma Culture". En Animal Cell Technology: Basic & Applied Aspects, 529–34. Dordrecht: Springer Netherlands, 1993. http://dx.doi.org/10.1007/978-94-011-2044-9_72.

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Actas de conferencias sobre el tema "Hybridoma cell"

Moffat, E. H., R. H. Furlong, A. L. Bloom y J. C. Giddings. "A MURINE MODEL FOR FACTOR VIII ANTIBODY ANTI-IDIOTYPE REAGENTS". En XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644030.

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The regulation of factor VIII antibody (FVIIIAb) production in haemophilic and non-haemophilic patients may be effected by anti-idiotype (Aid) antibodies which specifically react with FVIIIAb. Aid antibodies (reagents) were prepared from rabbits immunised with murine monoclonal FVIIIAb. Immuno fluorescent microscopy and cell culture studies were performed using murine hybridoma cells which secreted the FVIIIAbs.Immuno fluorescence studies examined the ability of the Aid reagents to bind to acetone fixed FVIIIAb secreting hybridoma cells. Positive surface membrane and intra-cytoplasmic staining patterns were seen with the Aid reagent when incubated with the corresponding murine hybridoma cell line. This reaction was blocked subsequent to the addition of the corresponding monoclonal FVIIIAb but was preserved when unrelated monoclonal FVIIIAb was incubated with the hybridoma cells. No fluorescence was observed when Aid reagent was incubated with unrelated FVIIIAb secreting hybridoma cultures.Following the addition of Aid reagent to FVIIIAb secreting hybridoma cultures and incubation for 19 hours, the resultant hybridoma supernatants were examined for FVIIIAb content using an immunoradiometric technique. The Aid reagent failed to inhibit FVIIIAb secretion by hybridoma cells. Thus, although Aid reagents were capable of binding to fixed FVIIIAb secreting cells, they failed to inhibit FVIIIAb secretion from hybridoma cultures. The conjugation of Aid reagent with immunotoxin may however have cytotoxic potential. The murine model provides a method for the study of Aid regulation of FVIIIAb production in haemophilia.

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Cruse-Sawyer, Janet E., B. Dixon, David J. Roberts, John Griffiths y Stanley B. Brown. "Photodynamic response of an endothelial hybridoma cell line using zinc(II) tetrasubstituted phthalocyanines". En Fifth International Photodynamic Association Biennial Meeting, editado por Denis A. Cortese. SPIE, 1994. http://dx.doi.org/10.1117/12.203449.

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Van Lent, Julie, Iene Rutten, Karen Ven, Jolien Breukers, Eleonore Verstraete, Katrien Van, Julie Van et al. "MICROFLUIDIC TOOLS FOR STUDYING SINGLE CELL SECRETIONS: A CASE STUDY ON HYBRIDOMA ANTIBODY SECRETION". En The 28th International Conference on Miniaturized Systems for Chemistry and Life Sciences - Micro-Total Analysis Systems. San Diego: Chemical and Biological Microsystems Society, 2024. https://doi.org/10.70477/zhun7496.

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Kitagawa, H., N. Yamamoto, G. Kosaki y H. Yamazaki. "AN IMPORTANT ROLE OF CARBOHYDRATE MOIETIES ON CANCER CELL MEMBRANE GLYCOPROTEINS IN CANCER CELL-INDUCED PLATELET AGGREGATION". En XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644667.

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Platelet aggregation induced by cancer cells may be an essential process in the development of hematogenous metastasis of cancers. A mechanism in HMV-I (human vaginal melanoma cell line)-induced platelet aggregation was studied by using monoclonal antibodies against membrane proteins of cancer cells or platelets. HMV-I cells or their membrana ractions induced platelet aggregation of human heparinized PRP, to which hirudin had no inhibitory effect. The platelet aggregation by HMV-I was completely lost after the pretreatment of the cells with 0.3U/ml neuraminidase for 60 min at 37°C. Preincubation of platelets with monoclonal antibodies against platelet GP lb or GP Ilb/lIIa inhibited HMV-I induced aggregation. A monoclonal antibody MB3 (igM) against another human melanoma (HMMB) which had been transplanted in nude mice was produced by hybridoma technique. Screening studies by cell binding ELISA revealed that MB3 antibody reacted with not only HMMB cells but also many other cells including HMV-I, M7609 (colon carcinoma cell line) and normal fibroblasts. Western-blot analyses showedthat MB3 antibody reacted with multiple, more than ten, proteins with molecular weights ranging from UO to 200 kDa in unreduced SDS-PAGE of HMV-I, HMMB or M7609. In contrast, when .these cells pretreated with neuraminidase were used in Western-blot, MB3 reactivity were all lost. MB3 reacted with at least three glycoproteins of human red cell membrane in Western-blot, but it did not react with human platelets. Immune adherent asgay with trypsin-treated HMV-I or HMMB cells as target cells showed negative reactivity. MB3 antibody inhibited HMV-I-induced aggregation of platelets, but did not inhibit M7609-induced aggregation which depended on thrombin generation.These results suggest that MB3 antibody may be against sialic acid-containing carbohydrate moieties of membrane glycoproteins on these cancercells and that the carbohydrate(s) may play a critical role in' cancer cell-platelet interaction.

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Chan, Leo L., Haohai Zhang, William Rice, Nasim Kassam, Maria S. Longhi, Haitao Zhao, Simon C. Robson, Wenda Gao y Yan Wu. "Abstract 5774: Novel cell-based high-throughput hybridoma screening method using the Celigo image cytometer for antibody discovery". En Proceedings: AACR Annual Meeting 2018; April 14-18, 2018; Chicago, IL. American Association for Cancer Research, 2018. http://dx.doi.org/10.1158/1538-7445.am2018-5774.

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Loureiro, Rafaele, José Senna y Álvaro Sousa. "Use of medium supplements to improve anti-MRSA mAb final concentration in hybridoma cell culture and reduce the cost production". En International Symposium on Immunobiologicals. Instituto de Tecnologia em Imunobiológicos, 2022. http://dx.doi.org/10.35259/isi.2022_52156.

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Sen, Sucharita y P. K. Roychoudhury. "Production of IgG1 monoclonal antibody 520C9 specific for human breast cancer oncoprotein c-erbB-2, using hybridoma cell line 8696 in perfusion bioreactor". En 3rd Annual International Conference on Advances in Biotechnology (BioTech 2013). Global Science and Technology Forum, 2013. http://dx.doi.org/10.5176/2251-2489_biotech13.25.

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Rahmani, A. F., S. Pambudi, S. A. Puteri, T. Subiantistha y R. Lestari. "Cloning of the heavy chain of fragment antigen binding anti-NS1 from hybridoma cell 71E2 induced by dengue virus on pTA2 vector in Escherichia coli TOP10". En PROCEEDINGS OF THE 4TH INTERNATIONAL SYMPOSIUM ON CURRENT PROGRESS IN MATHEMATICS AND SCIENCES (ISCPMS2018). AIP Publishing, 2019. http://dx.doi.org/10.1063/1.5132531.

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Haber, Edgar, Marchall T. Runge, Christoph Bode, Betsy Branscomb y Janet Schnee. "ANTIBODY TARGETED FIBRINOLYSIS". En XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643723.

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Chemical conjugates of fibrin-specificantibodies and plasminogen activators. Urokinase or tPA were linked covalently toamonoclonal antibody specific for the amino terminus of the beta chain of human fibrin (59D8) by means of the unidirectionalcross-linking reagent SPDP. The fibrinolytic potency of the conjugates at equal amidolytic activities was compared to the native plasminogen activators in an assay measuring lysis of 1251-fibrin monomer covalently linked to Sepharose CL-4B. Urokinase was least potent, tPA exhibited a 10fold increase in fibrinolysis whereas both the urokinase and tPA antibody conjugates and a urokinase-Fab conjugate were 250fold more potent than urokinase and 25 fold more potent than tPA. Enhanced fibrinolysis was fully inhibited by b peptide indicating its dependence on antigen binding. In a plasma assay conjugates of tPA orUK to antibody produced a 3.2- to 4.5-fold enhancement in clot lysis in human plasma over that of the respective unconjugated plasminogen activator. However, the UK-59D8 conjugate was only as potent as tPAalone. Antibody-conjugated tPA or UK consumed less fibrinogen, alpha 2-antiplasminand plasminogen than did the unconjugatedactivators, at equipotent thrombolytic concentrations. In a quantitative rabbit thrombolysis model, the activity of the purified conjugate was compared with that oftPA alone and that of a conjugate betweentPA and a digoxin-specific monoclonal antibody. After correction for spontaneous lysis, tPA-59D8 was shown to be 2.8 to,9.6times more potent than tPA alone. Unconjugated tPA and tPA-digoxin were equipotent.At equivalent thrombolytic concentrations, tPA-59D8 degraded less fibrinogen and consumed less alpha 2-antiplasmin than did tPA alone. These results suggest that tPA can be efficiently directed to the site of a thrombus by conjugation to an antifibrin monoclonal antibody, resulting in both more potent and more selective thrombolysis.A recombinant fusion protein comprising a fibrin-specific antibody site and theB chain of tPA. The rearranged 59D8 heavychain gene was cloned and combined in theexpression vector pSV2gpt withsequence coding for a portion of the Gamma 2b constant region and the catalytic beta chain of t-PA. This construct was transfected into heavy chain loss variant cells derived from the 59D8 hybridoma. Recombinant protein was purified by affinitychromatography and analyzed with Western blots. These revealed a 65-kD heavy chain-t-PA fusion protein that is secreted in association with the 59D8 light chain in the form of a 170-kD disulfide linked dimer. A chromogenic substrate assay showed the fusion protein to have 70 percent of the peptidolytic activity of native t-PA and to activate plasminogen as efficiently as t-PA. In a competitive binding assay, reconstituted antibody was shown to have a binding profile similar to that of native 59D8. Thus by recombinant techniques we have produced a novel hybrid protein capable of high affinity fibrin binding andplasminogen activation.Chemical conjugates between a fibrin-specific and a tPA-specific antibody. A heteroantibody duplex (duplex) with specificities for both tPA and fibrin was synthesized by conjugating iminothiolane-modified anti-tPA monoclonal antibody (TCL8) toantifibrin antibody 59D8. Addition of both duplex and tPA to a plasma clot assay gave more lysis (200 units produced 23.1 lysis; 400 units, 29.5 lysis) than did tPAalone (200 units, 1.8% lysis; 400 units,19% lysis). Despite increased potency associated with duplex addition, fibrinogen and alpha-2-antiplasmin levels at equal tPA concentrations did not differ. Thus, itis possible to concentrate tPA (added separately) to the site of a thrombus in plasma using a heteroantibody duplex with specificities for both tPA and fibrin.Biosynthetically produced heteroduplexantibodies that are both fibin and tPA-specific. The bispecific antibodies were prepared in two ways. First, polyethylene glycol-mediated fusions were performed with two different hybridoma cell lines: anti-fribrin b chain producer, 59D8 and anti-tPA producer, TCL8. TCL8 cells were selected for HPRT-minus variants and then fused with TK-deficient 59D8 cells. One cell line, F36.23, possessed both anti-human fibrin and anti-human t-PA immunoreactivities. A second method yielded another bispecific antibody, F32.1. This cell line was selected after fusing TCL8 (HPRT-minus) cells with spleen cells from a mouseimmunized with a fibrin-like peptide corresponding to the amino terminus of fibrinalpha-chains. Affinity-purified F32.1 andF36.23 retained anti-fibrin and anti-t-PAactivity and enhanced fibrinolytic potency of tPA by a factor of 10.

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Varon, D., S. Linder, E. Gembom, L. Guedj, A. Berrebi y Z. Eshhar. "MONOCLONAL ANTI-CYTOSKELETON ANTIBODY DERIVED FROM AN ITP PATIENT". En XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644581.

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A monoclonal anti-platelet IcpI antibody was established by the fusion of splenocytes from Immune Thrombocytopenic purpura (ITP) patient with a human-mouse-hetercmyelcma cell line. The splenic lynphocytes were cultured with platelets and lipopoly saccharide for 9 days prior to fusion. Anti-platelet activity in the hybridcma supernatant was tested by the ELISA technique using plastic adherent platelets as antigen. Out of three hybridomas that demonstrated anti-platelets activity, one (4G9) appeared to be stable producer of IgM, was cloned and served for further characterizations. In immunofluorescence studies it was found that 4G9 stains both platelets and mononuclear cells, exhibiting mainly intracellular pattern. When different cell lines were fixed and stained, specific decoration of cytoskeletal elements was detected on normal and transformed fibroblasts and not in epithelial or carcinoma cell lines. To characterize the antigen reacting with 4G9 antibody, whole platelet lysate was electrophoreased on SDS-PAGE, transferred to nitrocellulose filter and reacted with 4G9 followed by peroxidase anti-Ig staining. Major band of 45 Kd corresponding to act in, and minor band of 16 Kd reacted with 4G9 but not with control antibodies. The anti-actin activity of 4G9 was confirmed by immunoblot using purified rabbit-muscle actin, and by ELISA, using act in coated plates. Whether this autoantibody plays a role in the pathogenesis of ITP, or whether it belongs to the family of natural autoantibodies is under investigation.

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Informes sobre el tema "Hybridoma cell"

Jordan, Ramon L., Abed Gera, Hei-Ti Hsu, Andre Franck y Gad Loebenstein. Detection and Diagnosis of Virus Diseases of Pelargonium. United States Department of Agriculture, julio de 1994. http://dx.doi.org/10.32747/1994.7568793.bard.

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Pelargonium (Geranium) is the number one pot plant in many areas of the United States and Europe. Israel and the U.S. send to Europe rooted cuttings, foundation stocks and finished plants to supply a certain share of the market. Geraniums are propagated mainly vegetatively from cuttings. Consequently, viral diseases have been and remain a major threat to the production and quality of the crop. Among the viruses isolated from naturally infected geraniums, 11 are not specific to Pelargonium and occur in other crops while 6 other viruses seem to be limited to geranium. However, several of these viruses are not sufficiently characterized to conclude that they are distinct agents and their nomenclature and taxonomy are confusing. The ability to separate, distinguish and detect the different viruses in geranium will overcome obstacles te developing effective detection and certification schemes. Our focus was to further characterize some of these viruses and develop better methods for their detection and control. These viruses include: isolates of pelargonium line pattern virus (PLPV), pelargonium ringspot virus (PelRSV), pelargonium flower break virus (PFBV), pelargonium leaf curl (PLCV), and tomato ringspot virus (TomRSV). Twelve hybridoma cell lines secreting monoclonal antibodies specific to a geranium isolate of TomRSV were produced. These antibodies are currently being characterized and will be tested for the ability to detect TomRSV in infected geraniums. The biological, biochemical and serological properties of four isometric viruses - PLPV, PelRSV, and PFBV (and a PelRSV-like isolate from Italy called GR57) isolated from geraniums exhibiting line and ring pattern or flower break symptoms - and an isolate ol elderbeny latent virus (ELV; which the literature indicates is the same as PelRSV) have been determined Cloned cDNA copies of the genomic RNAs of these viruses were sequenced and the sizes and locations of predicted viral proteins deduced. A portion of the putative replicase genes was also sequenced from cloned RT-PCR fragments. We have shown that, when compared to the published biochemical and serological properties, and sequences and genome organizations of other small isometric plant viruses, all of these viruses should each be considered new, distinct members of the Carmovirus group of the family Tombusviridae. Hybridization assays using recombinant DNA probes also demonstrated that PLPV, PelRSV, and ELV produce only one subgenomic RNA in infected plants. This unusual property of the gene expression of these three viruses suggests that they are unique among the Carmoviruses. The development of new technologies for the detection of these viruses in geranium was also demonstrated. Hybridization probes developed to PFBV (radioactively-labeled cRNA riboprobes) and to PLPV (non-radioactive digoxigenin-labeled cDNAs) were generally shown to be no more sensitive for the detection of virus in infected plants than the standard ELISA serology-based assays. However, a reverse transcriptase-polymerase chain reaction assay was shown to be over 1000 times more sensitive in detecting PFBV in leaf extracts of infected geranium than was ELISA. This research has lead to a better understanding of the identity of the viruses infecting pelargonium and to the development of new tools that can be used in an improved scheme of providing virus-indexed pelargonium plants. The sequence information, and the serological and cloned DNA probes generated from this work, will allow the application of these new tools for virus detection, which will be useful in domestic and international indexing programs which are essential for the production of virus-free germplasm both for domestic markets and the international exchange of plant material.

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