Human antibodies and hybridomas最新文献_第5页

Phage displayed antibodies specific for a cytoskeletal antigen. Selection by competitive elution with a monoclonal antibody. 噬菌体显示针对细胞骨架抗原的特异性抗体。单克隆抗体竞争性洗脱选择。

Human antibodies and hybridomas Pub Date : 1995-01-01

E V Meulemans, L J Nieland, W H Debie, F C Ramaekers, G J van Eys

{"title":"Phage displayed antibodies specific for a cytoskeletal antigen. Selection by competitive elution with a monoclonal antibody.","authors":"E V Meulemans, L J Nieland, W H Debie, F C Ramaekers, G J van Eys","doi":"","DOIUrl":"","url":null,"abstract":"The display of repertoires of antibody fragments on the surface of filamentous phage offers a new way to produce immunoreagents with defined specificities. Here we report the selection of antibody fragments against the cytoskeletal fraction of T24 bladder cancer cells. To focus selection to a specific antigen, we eluted bound phage with a mouse monoclonal antibody directed against cytokeratins. Our initial studies proved that such a selection procedure with a library, carrying the mouse antibody fragment repertoire, resulted in phage specificity for the antigen against cytokeratin 8, recognized by the mouse monoclonal antibody. To facilitate detection of reactive clones, monoclonal antibodies against phage epitopes were developed. A human synthetic library (> 10(8) clones) was used for selection by competitive elution after binding to T24 cytoskeleton. About 50% of the phage reacted in ELISA with cytoskeletons of T24 cells, while with noncytokeratin containing cells no reaction was observed. Immunofluorescence studies and Western blotting with a number of these clones showed reactivity against cytokeratin. We conclude that the competitive elution method can be used as a rapid technique to obtain immunoreactive phages, and eventually human single chain antibodies directed against defined epitopes which were formerly characterized and validated by mouse monoclonal antibodies.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"6 3","pages":"113-8"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19577153","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Efficient immortalization of rheumatoid synovial tissue B-lymphocytes. A comparison between the techniques of electric field-induced and PEG fusion. 类风湿性滑膜组织b淋巴细胞的高效永生化。电场诱导与聚乙二醇融合技术之比较。

Human antibodies and hybridomas Pub Date : 1995-01-01 DOI: 10.3233/HAB-1995-6202

V. Krenn, P. von Landenberg, E. Woźniak, C. Kissler, H. Hermelink, U. Zimmermann, H. Vollmers

{"title":"Efficient immortalization of rheumatoid synovial tissue B-lymphocytes. A comparison between the techniques of electric field-induced and PEG fusion.","authors":"V. Krenn, P. von Landenberg, E. Woźniak, C. Kissler, H. Hermelink, U. Zimmermann, H. Vollmers","doi":"10.3233/HAB-1995-6202","DOIUrl":"https://doi.org/10.3233/HAB-1995-6202","url":null,"abstract":"In this study, B-cells isolated from rheumatoid synovial tissue were immortalized, without prior in vitro stimulation, by means of electric-field induced fusion and conventional PEG fusion in order to compare the efficiency of these methods. Two myeloma cell lines were used as fusion partners, the murine myeloma Ag8 and the murine-human heteromyeloma HAB-1. The results of seven fusion experiments performed simultaneously with identical cell populations showed that fusion frequencies obtained by electrofusion were 4 to 35 times higher than by the PEG fusion technique. The morphological and immunohistochemical evaluation of synovial tissues used for fusion showed that only tissues exhibiting a follicular distribution of B-cells with a high percentage of CD 22-positive lymphocytes gave rise to high fusion yields and produced B-cell clones, whereas synovial tissues with the same percentage of plasma cells but lower percentages of CD 22 lymphocytes yielded very low fusion rates. In conclusion, electrofusion is more efficient for immortalizing small amounts of synovial tissue B-lymphocytes than PEG fusion, since high fusion frequencies could be obtained by this technique without the need for prior in vitro stimulation. Synovial tissue exhibiting a follicular distribution of B-lymphocytes with high percentages of CD 22-positive lymphocytes gave rise to high hybridoma yields and therefore an ideal source of human rheumatoid B-cell clones.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"6 2 1","pages":"47-51"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/HAB-1995-6202","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69906041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 12

Comparison of anti-TNF alpha autoantibodies in plasma and from EBV transformed lymphocytes of autoimmune and normal individuals. 自身免疫与正常人血浆及EBV转化淋巴细胞抗tnf α自身抗体的比较。

Human antibodies and hybridomas Pub Date : 1995-01-01

L Tsuchiyama, T Wong, J Kieran, P Boyle, D Penza, G D Wetzel

引用次数: 0

Generation of a human monoclonal antibody to hepatitis C virus, JRA1 by activation of peripheral blood lymphocytes and hypo-osmolar electrofusion. 通过激活外周血淋巴细胞和低渗透电熔制备人丙型肝炎病毒单克隆抗体JRA1。

Human antibodies and hybridomas Pub Date : 1995-01-01 DOI: 10.3233/HAB-1995-6207

U. Zimmermann, L. Love-Homan, P. Gessner, D. Clark, G. Klöck, F. Johlin, G. Neil

{"title":"Generation of a human monoclonal antibody to hepatitis C virus, JRA1 by activation of peripheral blood lymphocytes and hypo-osmolar electrofusion.","authors":"U. Zimmermann, L. Love-Homan, P. Gessner, D. Clark, G. Klöck, F. Johlin, G. Neil","doi":"10.3233/HAB-1995-6207","DOIUrl":"https://doi.org/10.3233/HAB-1995-6207","url":null,"abstract":"We have generated a human monoclonal antibody with binding specificity for hepatitis C virus (HCV)-specific peptides using peripheral blood lymphocytes isolated from a HCV antibody positive patient. The B-lymphocytes were stimulated with lipopolysaccharide (LPS) for 72 hours prior to the fusion. A recently described high efficiency hypo-osmolar electrofusion technique was employed, allowing generation of a large number of human hybridomas. The hybridomas were screened for human immunoglobulin and HCV-specific peptide binding by EIA. A single HCV-positive clone, JRA1, was detected and sub-cloned. Isotype analysis showed it to secrete an IgM lambda monoclonal antibody. The antibody was positive on both first and second generation HCV antibody analysis. This study confirms that viable pathogen-specific B-cells may be recovered from the peripheral blood. Although such cells are likely to be relatively uncommon in the circulating B-cell pool, they may be successfully immortalized by high efficiency electrofusion techniques. This technique might be valuable for the generation of human monoclonal antibodies with specificity for other human pathogens.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"6 2 1","pages":"77-80"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/HAB-1995-6207","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69906139","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

A human-mouse chimeric Lym-1 monoclonal antibody with specificity for human lymphomas expressed in a baculovirus system. 一种在杆状病毒系统中表达的人类淋巴瘤特异性的人-小鼠嵌合Lym-1单克隆抗体。

Human antibodies and hybridomas Pub Date : 1995-01-01

P Hu, M S Glasky, A Yun, M M Alauddin, J L Hornick, L A Khawli, A L Epstein

{"title":"A human-mouse chimeric Lym-1 monoclonal antibody with specificity for human lymphomas expressed in a baculovirus system.","authors":"P Hu, M S Glasky, A Yun, M M Alauddin, J L Hornick, L A Khawli, A L Epstein","doi":"","DOIUrl":"","url":null,"abstract":"A murine anti-human B-cell monoclonal antibody, Lym-1, has shown considerable promise for the treatment of human malignant lymphomas and has been utilized as a new radioimmunotherapy for refractory lymphoma. In order to enhance its clinical potential, a genetically engineered chimeric Lym-1 (chLym-1) with murine variable regions and human gamma 1 and kappa constant regions was constructed and expressed. The goal of this study was to generate a Lym-1 reagent with decreased immunogenicity and improved effector functions. Murine Lym-1 variable region cDNAs were isolated from the murine Lym-1 hybridoma cell line, fused to gamma 1 and kappa constant region cDNAs, and expressed in an insect cell expression system with the baculovirus transfer vector pAcUW31. The chLym-1 antibody expressed in this system was correctly processed and assembled into the expected immunoglobulin monomer. Chimeric Lym-1 bound to both target antigen-bearing Raji cells and a Lym-1 anti-idiotype antibody and had a similar binding affinity as murine Lym-1. The chimeric and murine versions of Lym-1 were assayed for their ability to mediate antibody-dependent cellular cytotoxicity (ADCC) and to induce complement-mediated cytotoxicity (CMC) against Raji targets. Chimeric Lym-1 mediated a two-fold higher level of ADCC than murine Lym-1 and slightly lower levels of CMC than murine Lym-1. In addition, in Raji lymphoma-bearing nude mice, chLym-1 localized to the tumor with approximately equal uptake at 24 and 72 hours. Chimeric Lym-1, however, cleared from the blood of nontumor-bearing mice approximately 5 times faster than murine Lym-1 (20 h vs. 5 days), as expected for a xenogeneic protein. The improved in vitro and in vivo activities of this genetically engineered monoclonal antibody render it a new potential immunotherapeutic reagent for the treatment of human malignant lymphomas.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"6 2","pages":"57-67"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18500912","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Single-chain antibody streptavidin fusions: tetrameric bifunctional scFv-complexes with biotin binding activity and enhanced affinity to antigen. 单链抗体链亲和素融合物:具有生物素结合活性和增强抗原亲和力的四聚体双功能scfv复合物。

Human antibodies and hybridomas Pub Date : 1995-01-01

S M Kipriyanov, F Breitling, M Little, S Dübel

引用次数: 0

Induction of Goodpasture antibodies to noncollagenous domain (NC1) of type IV collagen in mice by idiotypic manipulation. 通过独特型操作诱导小鼠IV型胶原非胶原结构域(NC1)抗体。

Human antibodies and hybridomas Pub Date : 1995-01-01

Y Shoenfeld, B Gilburd, M Hojnik, M Damianovich, S Hacham, Y Kopolovic, P Polak-Charcon, I Goldberg, A Afek, L Hun-Chi

引用次数: 0

A human-mouse chimeric Lym-1 monoclonal antibody with specificity for human lymphomas expressed in a baculovirus system. 一种在杆状病毒系统中表达的人类淋巴瘤特异性的人-小鼠嵌合Lym-1单克隆抗体。

Human antibodies and hybridomas Pub Date : 1995-01-01 DOI: 10.3233/HAB-1995-6204

P. Hu, M. Glasky, A. Yun, M. Alauddin, J. Hornick, L. Khawli, A. Epstein

{"title":"A human-mouse chimeric Lym-1 monoclonal antibody with specificity for human lymphomas expressed in a baculovirus system.","authors":"P. Hu, M. Glasky, A. Yun, M. Alauddin, J. Hornick, L. Khawli, A. Epstein","doi":"10.3233/HAB-1995-6204","DOIUrl":"https://doi.org/10.3233/HAB-1995-6204","url":null,"abstract":"A murine anti-human B-cell monoclonal antibody, Lym-1, has shown considerable promise for the treatment of human malignant lymphomas and has been utilized as a new radioimmunotherapy for refractory lymphoma. In order to enhance its clinical potential, a genetically engineered chimeric Lym-1 (chLym-1) with murine variable regions and human gamma 1 and kappa constant regions was constructed and expressed. The goal of this study was to generate a Lym-1 reagent with decreased immunogenicity and improved effector functions. Murine Lym-1 variable region cDNAs were isolated from the murine Lym-1 hybridoma cell line, fused to gamma 1 and kappa constant region cDNAs, and expressed in an insect cell expression system with the baculovirus transfer vector pAcUW31. The chLym-1 antibody expressed in this system was correctly processed and assembled into the expected immunoglobulin monomer. Chimeric Lym-1 bound to both target antigen-bearing Raji cells and a Lym-1 anti-idiotype antibody and had a similar binding affinity as murine Lym-1. The chimeric and murine versions of Lym-1 were assayed for their ability to mediate antibody-dependent cellular cytotoxicity (ADCC) and to induce complement-mediated cytotoxicity (CMC) against Raji targets. Chimeric Lym-1 mediated a two-fold higher level of ADCC than murine Lym-1 and slightly lower levels of CMC than murine Lym-1. In addition, in Raji lymphoma-bearing nude mice, chLym-1 localized to the tumor with approximately equal uptake at 24 and 72 hours. Chimeric Lym-1, however, cleared from the blood of nontumor-bearing mice approximately 5 times faster than murine Lym-1 (20 h vs. 5 days), as expected for a xenogeneic protein. The improved in vitro and in vivo activities of this genetically engineered monoclonal antibody render it a new potential immunotherapeutic reagent for the treatment of human malignant lymphomas.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"11 5 1","pages":"57-67"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/HAB-1995-6204","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69906080","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 27

Induction of Goodpasture antibodies to noncollagenous domain (NC1) of type IV collagen in mice by idiotypic manipulation. 通过独特型操作诱导小鼠IV型胶原非胶原结构域(NC1)抗体。

Human antibodies and hybridomas Pub Date : 1995-01-01 DOI: 10.3233/HAB-1995-6401

Y. Shoenfeld, B. Gilburd, M. Hojnik, M. Damianovich, S. Hacham, Y. Kopolovic, P. Polak-Charcon, I. Goldberg, A. Afek, L. Hun-Chi

引用次数: 7

A genetically engineered fusion protein M4/TNF with increased bifunctional activity refolded in the presence of protein disulfide isomerase. 具有增加双功能活性的基因工程融合蛋白M4/TNF在蛋白二硫异构酶存在下重新折叠。

Human antibodies and hybridomas Pub Date : 1995-01-01 DOI: 10.3233/HAB-1995-6402

J. Yang, R. Raju, R. Sharma, J. Xiang

引用次数: 5