Human antibodies and hybridomas最新文献_第10页

Human monoclonal antibodies recognize early and late viral proteins of human cytomegalovirus. 人单克隆抗体识别人巨细胞病毒的早期和晚期病毒蛋白。

Human antibodies and hybridomas Pub Date : 1994-01-01 DOI: 10.3233/HAB-1994-51-211

H. Alexander, J. Harpprecht, H. -. Podzuweit, P. Rautenberg, W. Müller‐ruchholtz

{"title":"Human monoclonal antibodies recognize early and late viral proteins of human cytomegalovirus.","authors":"H. Alexander, J. Harpprecht, H. -. Podzuweit, P. Rautenberg, W. Müller‐ruchholtz","doi":"10.3233/HAB-1994-51-211","DOIUrl":"https://doi.org/10.3233/HAB-1994-51-211","url":null,"abstract":"Human monoclonal antibodies directed against human cytomegalovirus were generated by fusion of in vitro stimulated human spleen lymphocytes from an HCMV-seropositive 53-year-old organ donor with the mouse myeloma cell line Ag8.653. Fourteen human/mouse hybridomas producing anti-cytomegalovirus IgG were screened by an ELISA technique and four selected clones have been established since March 1988, generating about 5-40 micrograms/24 h IgG per ml culture supernatant. Reference and local cytomegalovirus strains were stained by the antibodies without showing cross-reactivity to other herpes viruses. Three monoclonal antibodies, A4B4 (IgG11), A6B3 (IgG1k) and A6A2 (IgG1k), immunoprecipitated a 68 kDa early viral protein which appears during the infectious cycle, first in the nucleus (18-24 h) and then also in the cytoplasm (24-96 h) of infected cells. Inhibition of DNA replication restricted the detection of the 68 kDa viral protein to the nucleus of infected cells. Staining of unfixed infected cells showed that two of the antibodies bound at the surface of a few cells. The fourth monoclonal antibody A3C5 (IgG11) immunoprecipitated a 34/38 kDa late viral protein which appears in the nucleus (48-72 h) of infected cells. These antibodies enable us to study the human host response to human cytomegalovirus and to elucidate the functions of human antibodies especially in their interaction with the T-cell response.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"5 1-2 1","pages":"81-90"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/HAB-1994-51-211","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69905979","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: 5

Construction and characterization of chimeric and humanized forms of a broadly neutralizing monoclonal antibody to HIV-1. 一种广泛中和型HIV-1单克隆抗体的嵌合和人源化形式的构建和表征。

Human antibodies and hybridomas Pub Date : 1994-01-01 DOI: 10.3233/HAB-1994-51-202

J. Major, R. Liou, L. Sun, L. Yu, S. Starnes, M. Fung, T. Chang, N. Chang

{"title":"Construction and characterization of chimeric and humanized forms of a broadly neutralizing monoclonal antibody to HIV-1.","authors":"J. Major, R. Liou, L. Sun, L. Yu, S. Starnes, M. Fung, T. Chang, N. Chang","doi":"10.3233/HAB-1994-51-202","DOIUrl":"https://doi.org/10.3233/HAB-1994-51-202","url":null,"abstract":"Murine monoclonal antibody (MAb) G3-519 has been shown to recognize a conserved neutralizing epitope in the fourth constant (C4) region of the external glycoprotein gp120 of HIV-1. Inasmuch as this antibody effectively neutralized the infectivity of diverse HIV-1 isolates, it has been selected to be developed for passive immunization against HIV-1 infection in humans. In order to minimize the problem of immunogenicity of murine antibodies and to confer additional accessory immune functions, we have constructed mouse/human chimeric and humanized forms of the antibody. The chimeric antibody was constructed by cloning the murine variable regions and replacing the mouse constant regions with those from human Ig gamma 1,kappa. The humanized antibody was constructed using the human KAS variable region framework sequences as template. Engineering was guided by a three dimensional model of the murine variable region. The murine, chimeric and humanized forms of the antibody exhibited similar reactivity with the peptidic antigen in ELISA, and comparably neutralized the infectivity of HIV-1 in vitro. Taken together, our results show that the chimeric and humanized forms of G3-519 essentially retain the binding activity of the mouse parental antibody. Clinical development is planned to assess the prophylactic and therapeutic usefulness of these reshaped antibodies in humans.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"5 1-2 1","pages":"9-17"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/HAB-1994-51-202","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69906234","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: 3

Design and analysis of PCR primers for the amplification and cloning of human immunoglobulin Fab fragments. 人免疫球蛋白Fab片段PCR引物的设计与分析。

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

M de Boer, S Y Chang, G Eichinger, H C Wong

引用次数: 0

Human monoclonal Fab fragments from a combinatorial library prepared from an individual with a low serum titer to a virus. 从对病毒具有低血清效价的个体制备的组合文库中的人单克隆Fab片段。

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

E Bender, G R Pilkington, D R Burton

引用次数: 0

Comparison of expression of a humanized monoclonal antibody in mouse NSO myeloma cells and Chinese hamster ovary cells. 人源化单克隆抗体在小鼠NSO骨髓瘤细胞和中国仓鼠卵巢细胞中的表达比较。

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

T C Peakman, J Worden, R H Harris, H Cooper, J Tite, M J Page, D R Gewert, M Bartholemew, J S Crowe, S Brett

{"title":"Comparison of expression of a humanized monoclonal antibody in mouse NSO myeloma cells and Chinese hamster ovary cells.","authors":"T C Peakman, J Worden, R H Harris, H Cooper, J Tite, M J Page, D R Gewert, M Bartholemew, J S Crowe, S Brett","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>We report a detailed comparison of two commonly used stable, amplifiable mammalian expression systems (Chinese Hamster Ovary cells/dihydrofolate reductase and Mouse NSO myeloma/glutamine synthetase) used to express a humanized IgG1 monoclonal antibody. We compare copy number and steady state mRNA levels of both the selectable marker and heavy chain of the antibody throughout the selection and amplification process. In both cell lines, copy number and steady state levels of heavy chain and selectable marker increased during selection and were further increased during amplification. As expected, an increase in steady state mRNA levels of heavy chain correlated with an increase in expression of antibody whilst an increase in the steady state levels of mRNA of the selectable marker correlated with increased resistance to the selective agent. In NSO and CHO cells producing equivalent amounts of antibody, the copy number of the antibody genes and selectable marker was significantly higher in the CHO cells than in the NSO cells. However, the steady state mRNA levels of the heavy chain of the antibody were virtually identical. Rates of protein secretion in the two cell lines were also compared and found to be very similar. When the antibody purified from both systems was compared in a number of functional assays they behaved identically.</p>","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"5 1-2","pages":"65-74"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18539622","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

Anti-glycolipid antibodies produced by Epstein-Barr virus (EBV)-transformed oligoclonal B cell lines obtained from normal persons and autoimmune disease patients. 从正常人和自身免疫性疾病患者获得的eb病毒(EBV)转化寡克隆B细胞系产生抗糖脂抗体

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

Y Nagatsuka, S Hanazawa, T Yasuda, Y Ono

引用次数: 0

Specific reactivity of human monoclonal antibody AE6F4 against cancer cells in tissues and sputa from lung cancer patients. 人单克隆抗体AE6F4对肺癌患者组织及痰液中癌细胞的特异性反应性。

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

M Shoji, S Kawamoto, S Sato, M Kamei, M Kato, S Hashizume, K Seki, K Yasumoto, A Nagashima, H Nakahashi

引用次数: 0

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 DOI: 10.3233/HAB-1994-51-203

N. Heveker, A. Hansen, K. Hungerer, R. von Baehr, R. Glaser

{"title":"A human monoclonal antibody with the capacity to neutralize Staphylococcus aureus alpha-toxin.","authors":"N. Heveker, A. Hansen, K. Hungerer, R. von Baehr, R. Glaser","doi":"10.3233/HAB-1994-51-203","DOIUrl":"https://doi.org/10.3233/HAB-1994-51-203","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 1","pages":"18-24"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/HAB-1994-51-203","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69906243","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: 3

Anti-Tn human monoclonal antibodies generated following active immunization with partially desialylated ovine submaxillary mucin. 部分脱氮羊颌下黏液蛋白主动免疫后产生的抗tn人单克隆抗体。

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

K P O'Boyle, K Wright

引用次数: 0

Human monoclonal anti-Rh antibodies produced by human-mouse heterohybridomas express the Gal alpha 1-3 Gal epitope. 人小鼠异杂交瘤产生的人单克隆抗rh抗体表达Gal α 1-3 Gal表位。

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

R F Montaño, E L Romano

引用次数: 0