Human antibodies and hybridomas最新文献_第7页

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

U Zimmermann, L Love-Homan, P Gessner, D Clark, G Klöck, F C Johlin, G A 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 C Johlin, G A Neil","doi":"","DOIUrl":"","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","pages":"77-80"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18500915","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 DOI: 10.3233/HAB-1995-6303

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

引用次数: 32

Antigen-specific primary immune response of human B-lymphocytes after in vitro immunization with GM3 ganglioside. GM3神经节苷脂体外免疫后人b淋巴细胞抗原特异性初级免疫应答的研究。

Human antibodies and hybridomas Pub Date : 1995-01-01 DOI: 10.3233/HAB-1995-6304

M. Alfonso, B. Lanne, P. Ifversen, A. Vázquez, R. Pérez, J. Portoukalian, J. Zeuthen

{"title":"Antigen-specific primary immune response of human B-lymphocytes after in vitro immunization with GM3 ganglioside.","authors":"M. Alfonso, B. Lanne, P. Ifversen, A. Vázquez, R. Pérez, J. Portoukalian, J. Zeuthen","doi":"10.3233/HAB-1995-6304","DOIUrl":"https://doi.org/10.3233/HAB-1995-6304","url":null,"abstract":"In vitro immunization of human B-lymphocytes was performed with liposomes containing the monosialoganglioside GM3, with or without either complete tetanus toxoid or a synthetic T helper epitope derived from tetanus toxin (determinant 830-843). The immunized B-cells were Epstein-Barr virus transformed and the human anti-ganglioside antibody response was evaluated using an indirect ELISA against different mono- and disialogangliosides. Clones producing antigen-specific human antibodies of the IgM isotype against the ganglioside GM3 used as the immunogen were selected and one clone, IM-11, was further characterized. In addition, a method of positive selection using GM3-coated magnetic beads has been developed which allowed us to rescue unstable clones. The binding of the human antibody IM-11 to a large panel of glycosphingolipids separated on thin-layer plates was studied. The human MAb IM-11 was found to bind strongly to NeuAcGM3, IV3 NeuAcnLc4 and sulfate containing glycosphingolipids and weakly to NeuGcGM3. Immunohistological staining of melanoma and breast cancer biopsy sections showed a selective reactivity of IM-11 with tumor cells which varied among different tumors.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"6 3 1","pages":"102-12"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/HAB-1995-6304","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69906272","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: 6

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

J Yang, R Raju, R Sharma, J Xiang

引用次数: 0

Immunocytochemical detection of lung cancer cells with monoclonal antibodies to 14-3-3 proteins. 肺癌细胞14-3-3蛋白单克隆抗体的免疫细胞化学检测。

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

Y Setoguchi, M Kato, M Shoji, T Honjoh, M Kamei, M Sugitani, S Sato, S Hashizume, T Hanagiri, T Yoshimatsu

{"title":"Immunocytochemical detection of lung cancer cells with monoclonal antibodies to 14-3-3 proteins.","authors":"Y Setoguchi, M Kato, M Shoji, T Honjoh, M Kamei, M Sugitani, S Sato, S Hashizume, T Hanagiri, T Yoshimatsu","doi":"","DOIUrl":"","url":null,"abstract":"Murine monoclonal antibodies were raised against 14-3-3 proteins, the antigen of human monoclonal antibody AE6F4 which had been shown potentially useful for the immunochemical diagnosis of lung cancer via sputum cytology. Enzyme-linked immunosorbent assays of the murine anti-14-3-3 monoclonal antibodies with isolated bovine brain 14-3-3 isoforms showed that the antibodies were classified into four different profiles of isoform reactivity. The comparison of 14-3-3 isoform and lung cancer tissue on the reactivity with murine monoclonal antibodies indicated that beta isoform can be responsible for cancer recognition, whereas human monoclonal antibody AE6F4 showed preferential binding to zeta isoform. No murine monoclonal antibody of the same isoform specificity as human monoclonal antibody AE6F4 was obtained. Since murine monoclonal antibodies with different isoform specificities could immunostain lung cancer cells in sputum successfully, the combination use of murine monoclonal anti-14-3-3 antibodies with human monoclonal antibody AE6F4 is potentially useful for facilitating the sputum cytodiagnosis of lung cancer.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"6 4","pages":"137-44"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19663089","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

Comparison of three human-murine heteromyeloma cell lines for formation of human hybridomas after electrofusion with human peripheral blood lymphocytes from meningococcal cases and carriers. 三种人鼠异骨髓瘤细胞系与脑膜炎球菌病例和携带者外周血淋巴细胞电融合后形成人杂交瘤的比较。

Human antibodies and hybridomas Pub Date : 1995-01-01 DOI: 10.3233/HAB-1995-6201

A. Delvig, B. Koumaré, R. Glaser, J. Wang, S. Jahn, M. Achtman

引用次数: 2

The biological activity of human monoclonal IgG anti-D is reduced by beta-galactosidase treatment. 经-半乳糖苷酶处理后，人单克隆IgG抗d的生物活性降低。

Human antibodies and hybridomas Pub Date : 1995-01-01 DOI: 10.3233/HAB-1995-6301

B. Kumpel, Yan Wang, Helen L. Griffiths, Andrew G. Hadley, Graham A. W. Rook

{"title":"The biological activity of human monoclonal IgG anti-D is reduced by beta-galactosidase treatment.","authors":"B. Kumpel, Yan Wang, Helen L. Griffiths, Andrew G. Hadley, Graham A. W. Rook","doi":"10.3233/HAB-1995-6301","DOIUrl":"https://doi.org/10.3233/HAB-1995-6301","url":null,"abstract":"The contribution to IgG effector function of exposed galactose residues on the oligosaccharide chains in IgG has been tested experimentally. We studied a human monoclonal antibody (BRAD-5) to the Rh D blood group antigen. The preparation used contained a very low percentage of agalactosyl IgG (3.6%). After digestion with beta-galactosidase there was an increase in terminal GlcNAc indicating an increase in % agalactosyl IgG to approximately 30%. Comparison was made of the Fc receptor-(Fc gamma R)-mediated functional interactions of the two glycoforms of BRAD-5 in assays which measured the recognition and destruction of sensitised erythrocytes by various effector cells. After beta-galactosidase treatment, there was a slight reduction in Fc gamma RI-mediated adherence of erythrocytes to U937 cells and phagocytosis of erythrocytes by monocytes. Fc gamma RII-mediated binding of erythrocytes to K562 cells was also reduced. However there was little difference in adherence of erythrocytes to either Daudi cells (via Fc gamma RII) or NK cells (via Fc gamma RIII). There was a consistent reduction in lysis of erythrocytes mediated through Fc gamma RIII on K cells with the beta-galactosidase treated anti-D. Overall the results showed that reduced levels of galactose on IgG anti-D were associated with reduced biological activity in these assays, but the experiments failed to cast light on the physiological role of the agalactosyl glycoform of IgG.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"6 3 1","pages":"82-8"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/HAB-1995-6301","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69906195","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: 74

Immunocytochemical detection of lung cancer cells with monoclonal antibodies to 14-3-3 proteins. 肺癌细胞14-3-3蛋白单克隆抗体的免疫细胞化学检测。

Human antibodies and hybridomas Pub Date : 1995-01-01 DOI: 10.3233/HAB-1995-6403

Y. Setoguchi, M. Kato, M. Shoji, T. Honjoh, M. Kamei, M. Sugitani, S. Sato, S. Hashizume, T. Hanagiri, T. Yoshimatsu

{"title":"Immunocytochemical detection of lung cancer cells with monoclonal antibodies to 14-3-3 proteins.","authors":"Y. Setoguchi, M. Kato, M. Shoji, T. Honjoh, M. Kamei, M. Sugitani, S. Sato, S. Hashizume, T. Hanagiri, T. Yoshimatsu","doi":"10.3233/HAB-1995-6403","DOIUrl":"https://doi.org/10.3233/HAB-1995-6403","url":null,"abstract":"Murine monoclonal antibodies were raised against 14-3-3 proteins, the antigen of human monoclonal antibody AE6F4 which had been shown potentially useful for the immunochemical diagnosis of lung cancer via sputum cytology. Enzyme-linked immunosorbent assays of the murine anti-14-3-3 monoclonal antibodies with isolated bovine brain 14-3-3 isoforms showed that the antibodies were classified into four different profiles of isoform reactivity. The comparison of 14-3-3 isoform and lung cancer tissue on the reactivity with murine monoclonal antibodies indicated that beta isoform can be responsible for cancer recognition, whereas human monoclonal antibody AE6F4 showed preferential binding to zeta isoform. No murine monoclonal antibody of the same isoform specificity as human monoclonal antibody AE6F4 was obtained. Since murine monoclonal antibodies with different isoform specificities could immunostain lung cancer cells in sputum successfully, the combination use of murine monoclonal anti-14-3-3 antibodies with human monoclonal antibody AE6F4 is potentially useful for facilitating the sputum cytodiagnosis of lung cancer.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"2 1","pages":"137-44"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/HAB-1995-6403","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69906408","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: 8

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 DOI: 10.3233/HAB-1995-6305

E. Meulemans, L. Nieland, W. Debie, F. Ramaekers, G. V. van Eys

{"title":"Phage displayed antibodies specific for a cytoskeletal antigen. Selection by competitive elution with a monoclonal antibody.","authors":"E. Meulemans, L. Nieland, W. Debie, F. Ramaekers, G. V. van Eys","doi":"10.3233/HAB-1995-6305","DOIUrl":"https://doi.org/10.3233/HAB-1995-6305","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 1","pages":"113-8"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/HAB-1995-6305","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69906332","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: 9

A human monoclonal antibody with the capacity to neutralize Staphylococcus aureus alpha-toxin. 一种人单克隆抗体，具有中和金黄色葡萄球菌α毒素的能力。

Human antibodies and hybridomas Pub Date : 1994-01-01

N Heveker, A Hansen, K D Hungerer, R von Baehr, R W Glaser

{"title":"A human monoclonal antibody with the capacity to neutralize Staphylococcus aureus alpha-toxin.","authors":"N Heveker, A Hansen, K D Hungerer, R von Baehr, R W Glaser","doi":"","DOIUrl":"","url":null,"abstract":"A human monoclonal antibody, CB STL-1, against staphylococcal alpha-toxin has been established by hybridoma technology. It is of IgG1 subclass with lambda light chain and possesses a dissociation constant of 8 x 10(-10) mol/l. 1 mg of purified antibody neutralizes the hemolytic activity of 800 micrograms/ml alpha-toxin in an in vitro hemolysis assay using rabbit erythrocytes. The antibody does not bind to overlapping (7 residues) decapeptides spanning the sequence of alpha toxin, thus it might bind to a conformational epitope. The epitope recognized by the antibody is not accessible in oligomeric toxin. The antibody binds both to the hydrophilic and amphipathic forms of the monomeric toxin Fab fragments of the antibody are stable and show no significant loss of activity. CB STL-1 was able to protect mice in vivo from i.p. challenge with alpha toxin. Thus, the antibody is a candidate for passive immunotherapy. The variable regions of the antibody secreted by CB STL-1 were sequenced and found to be encoded by a VH gene segment belonging to the VH1 family, and a Vlambda segment most likely belonging to the VlambdaIII subgroup. Further analysis concerning the third complementarity determining region (CDR3) of the heavy chain is presented.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"5 1-2","pages":"18-24"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18855961","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