Hybridoma最新文献_第10页

Induction of monoclonal antibody with predefined ELNKWA epitope specificity by epitope vaccine. 表位疫苗诱导具有预定义ELNKWA表位特异性的单克隆抗体。

Hybridoma Pub Date : 2000-08-01 DOI: 10.1089/027245700429918

Y Xiao, X N Dong, Y H Chen

{"title":"Induction of monoclonal antibody with predefined ELNKWA epitope specificity by epitope vaccine.","authors":"Y Xiao, X N Dong, Y H Chen","doi":"10.1089/027245700429918","DOIUrl":"https://doi.org/10.1089/027245700429918","url":null,"abstract":"Since the hybridoma technique to produce monoclonal antibodies (MAbs) was discovered, thousands of MAbs with predefined protein specificity have been produced, and a natural or recombinant protein as antigen is necessary for inducing MAbs in the conventional hybridoma technique. To induce epitope-specific MAbs, we suggest an epitope vaccine as a new technique to induce MAbs with predefined epitope specificity. ELDKWA was identified as an important neutralizing epitope on HIV-1 gp41. The MAb 2F5, recognizing ELDKWA epitope, has shown broad neutralizing activity to many HIV strains, including primary isolates, but the mutant in ELNKWA epitope results in escape 2F5-based neutralization. To produce MAbs recognizing this mutated epitope for consideration of passive immunotherapy against the mutant bearing the ELNKWA epitope, MAbs with predefined ELNKWA epitope specificity were induced by synthetic epitope-peptide instead of a natural or recombinant gp41 bearing this epitope. Three MAbs were identified to recognize ELNKWA epitope on the synthetic epitope-peptide, and interestingly could bind the recombinant gp41 with ELDKWA epitope in an ELISA assay and immunoblotting analysis.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"19 4","pages":"347-50"},"PeriodicalIF":0.0,"publicationDate":"2000-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245700429918","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21834661","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

Generation of monoclonal antibodies directed against the immunogenic glycoprotein K8.1 of human herpesvirus 8. 人疱疹病毒8免疫原糖蛋白K8.1单克隆抗体的制备。

Hybridoma Pub Date : 2000-08-01 DOI: 10.1089/027245700429837

D Lang, A Birkmann, F Neipel, W Hinderer, M Rothe, M Ernst, H H Sonneborn

{"title":"Generation of monoclonal antibodies directed against the immunogenic glycoprotein K8.1 of human herpesvirus 8.","authors":"D Lang, A Birkmann, F Neipel, W Hinderer, M Rothe, M Ernst, H H Sonneborn","doi":"10.1089/027245700429837","DOIUrl":"https://doi.org/10.1089/027245700429837","url":null,"abstract":"Human Herpesvirus 8 (HHV-8) is clearly associated with Kaposi's sarcoma (KS), body cavity-based lymphomas (BCBL), and certain forms of multifocal Castleman's disease (MCD). It appears to be the sexually transmissible agent involved in the development of AIDS-associated KS. HHV-8 genomes are invariably present in BCBL-derived cell lines where lytic replication of the virus can be induced by phorbol esters (PE). First-generation HHV-8 serological assays were based on these cell lines. More recently, several genes encoding HHV-8 antigens have been identified. One of the most reactive antigens is encoded by HHV-8 open reading frame K8.1. Although K8.1 does not exhibit overt sequence homology to any other known gene, it is likely to be analogous to gp220/350 of Epstein-Barr or gp150 of murine herpesvirus-68, virion-envelope glycoproteins involved in target cell recognition. Mice were immunized with purified GST-K8.1 fusion protein expressed in E. coli. After fusion of murine plasma cells with the myeloma cell line P3-X63-Ag8. monoclonal antibodies (MAbs) were generated, which are specifically directed against K8.1 protein. The binding site for each MAb was identified by deletion mutant analysis using recombinant GST-K8.1 mutants and K8.1-specific peptides. Without exception, the epitopes recognized by these MAbs were located within the N-terminal part of the protein [amino acids (aa) 29 to 80], thus identifying a highly immunogenic region. These antibodies will not only be useful tools for HHV-8 diagnostics, but will also facilitate the analysis of K8.1 function.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"19 4","pages":"287-95"},"PeriodicalIF":0.0,"publicationDate":"2000-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245700429837","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21834050","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

Production and characterization of specific monoclonal antibodies of the human thyroid stimulating hormone. 人促甲状腺激素特异性单克隆抗体的制备与鉴定。

Hybridoma Pub Date : 2000-08-01 DOI: 10.1089/027245700429891

I B Baluja Conde, A I Brito Moreno, I Amores Sanchéz, C Acosta Bas

引用次数: 4

Monoclonal antibodies generated against recombinant ATM support kinase activity. 生成的抗重组ATM的单克隆抗体支持激酶活性。

Hybridoma Pub Date : 2000-08-01 DOI: 10.1089/027245700429864

K J Alligood, M Milla, N Rhodes, B Ellis, K E Kilpatrick, A Lee, T M Gilmer, T J Lansing

引用次数: 9

Veterinary sources of nonrodent monoclonal antibodies: interspecific and intraspecific hybridomas. 非啮齿动物单克隆抗体的兽医来源:种间和种内杂交瘤。

Hybridoma Pub Date : 2000-06-01 DOI: 10.1089/02724570050109602

D J Groves, B A Morris

引用次数: 35

Isolation and characterization of monoclonal antibodies specific for protein conformational epitopes present in prostate-specific membrane antigen (PSMA). 前列腺特异性膜抗原(PSMA)蛋白构象表位特异性单克隆抗体的分离和鉴定。

Hybridoma Pub Date : 2000-06-01 DOI: 10.1089/02724570050109648

W T Tino, M J Huber, T P Lake, T G Greene, G P Murphy, E H Holmes

{"title":"Isolation and characterization of monoclonal antibodies specific for protein conformational epitopes present in prostate-specific membrane antigen (PSMA).","authors":"W T Tino, M J Huber, T P Lake, T G Greene, G P Murphy, E H Holmes","doi":"10.1089/02724570050109648","DOIUrl":"https://doi.org/10.1089/02724570050109648","url":null,"abstract":"Prostate-specific membrane antigen (PSMA) is a 750-amino acid glycoprotein highly expressed in malignant prostate tissues. PSMA reacts with the murine monoclonal antibody 7E11.C5, whose binding epitope has been mapped to the N-terminal of the protein distributed on the cytoplasmic side of the plasma membrane. We have developed murine monoclonal antibodies specific for extracellular epitopes of PSMA. Three of these antibodies--1G9, 3C6, and 4D4--display distinct binding properties consistent with their recognition of conformational epitopes within native PSMA. Results indicate this panel of antibodies binds to native full-length PSMA, but not to fusion proteins containing portions of the linear sequence of the protein. Antibody binding is greatly reduced upon heat denaturation of native PSMA, and these antibodies do not detect PSMA by Western blot. Immunoprecipitation experiments demonstrate the ability of each to bind to full-length PSMA as well as PSM', a form of the protein missing the first 57 amino acids. These results indicate each antibody is specific for an epitope within the extracellular domain, a region spanning residues 44-750. Flow cytometric experiments indicate strong specific binding to live LNCaP cells. Antibody inhibition studies demonstrate that these antibodies recognize at least two distinct epitopes. Taken together, the results demonstrate that these antibodies are specific for native protein conformational epitopes within the extracellular domain. Their properties, in particular strong binding to live cancer cells, make them ideal candidates that are clearly superior to linear sequence epitope specific antibodies for in vivo applications.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"19 3","pages":"249-57"},"PeriodicalIF":0.0,"publicationDate":"2000-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/02724570050109648","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21790450","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

Antibody binding regions on human nerve growth factor identified by homolog- and alanine-scanning mutagenesis. 用同源和丙氨酸扫描诱变法鉴定人神经生长因子抗体结合区。

Hybridoma Pub Date : 2000-06-01 DOI: 10.1089/02724570050109611

J S Hongo, G R Laramee, R Urfer, D L Shelton, T Restivo, M Sadick, A Galloway, H Chu, J W Winslow

{"title":"Antibody binding regions on human nerve growth factor identified by homolog- and alanine-scanning mutagenesis.","authors":"J S Hongo, G R Laramee, R Urfer, D L Shelton, T Restivo, M Sadick, A Galloway, H Chu, J W Winslow","doi":"10.1089/02724570050109611","DOIUrl":"https://doi.org/10.1089/02724570050109611","url":null,"abstract":"The binding specificities of a panel of mouse monoclonal antibodies (MAbs) to human nerve growth factor (hNGF) were determined by epitope mapping using chimeric and point mutants of NGF. Subsequently, the MAbs were used to probe NGF structure-function relationships. Six MAbs, which recognize distinct or partially overlapping regions of hNGF, were evaluated for their ability to block the binding of hNGF to the TrkA and p75 NGF receptors in various in vitro assays, which included blocking of TrkA autophosphorylation and blocking of NGF-dependent survival of dorsal root ganglion sensory neurons. Three MAbs (911,912,938) were potent blockers of all activities. Potent blocking of p75 binding occurs only with MAb 909, which recognizes an NGF region identified by mutagenesis as important for NGF-p75 binding. These results are consistent with recently proposed models of binding regions involved in NGF-TrkA and NGF-p75 interactions generated through mutagenic analysis and structure determination of the NGF-TrkA complex. These studies provide insight to the epitope specificities and potency of MAbs that would be useful for physiological NGF blocking studies.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"19 3","pages":"215-27"},"PeriodicalIF":0.0,"publicationDate":"2000-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/02724570050109611","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21790506","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: 37

Lewis antigens and argyrophilic nucleolar organizer regions staining for assessment of potential malignancy of adenomatous polyps of the gastrointestinal tract in children. Lewis抗原和亲嗜嗜核仁组织区染色评估儿童胃肠道腺瘤性息肉的潜在恶性。

Hybridoma Pub Date : 2000-06-01 DOI: 10.1089/02724570050109675

W Romańczuk, K Steplewska-Mazur, B M Woźniewicz, R Korczowski

{"title":"Lewis antigens and argyrophilic nucleolar organizer regions staining for assessment of potential malignancy of adenomatous polyps of the gastrointestinal tract in children.","authors":"W Romańczuk, K Steplewska-Mazur, B M Woźniewicz, R Korczowski","doi":"10.1089/02724570050109675","DOIUrl":"https://doi.org/10.1089/02724570050109675","url":null,"abstract":"Adenomatous polyps (AP) of the gastrointestinal tract in children are very rare. Because of their potential malignancy, they are of great clinical importance. There is little experience in the management of children with AP. The immunohistochemical expression of the Lewis blood group antigens (BGA) (sialosyl-Le(a), Le(a), Leb, Le(x), and Le(y)) and the number of activated nucleoli with the silver staining method for nucleolar organizer regions (AgNORs) were studied in two children with AP. In a girl with isolated AP of the stomach and colon, it was found that antigens Le(b) and s-Le(a) were expressed extensively in the gastric adenoma, and sialosyl-Le(a) throughout the entire length of the rectal adenoma crypts, but in the AgNORs stain the number of nucleoli ranged from two to four, evidencing changes of a benign character. In the case of familial adenomatous polyposis diagnosed in a 9-year-old boy, in some colonic adenomas the number of activated nucleoli was greater than five, and the Le(b) antigen was expressed in superficial epithelial cells in one of the adenomas. Also, extensive expression of antigens Le(y) and s-Le(a) throughout the entire length of the crypt in another polyp removed was observed. We believe that immunohistochemical study of the intensity and extent of the expression of Lewis BGA in the polyp tissue simultaneously with the determination of the number of activated nucleoli by the AgNORs staining method can be helpful in better analysis of cytological risk factors of a malignant transformation.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"19 3","pages":"269-76"},"PeriodicalIF":0.0,"publicationDate":"2000-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/02724570050109675","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21790453","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

14F7 Anti-GM3(NeuGc)ganglioside. 14 f7 Anti-GM3神经节苷脂(NeuGc)。

Hybridoma Pub Date : 2000-06-01 DOI: 10.1089/02724570050109684

引用次数: 0

2F12-H9; 2C4-B2; 2C4-A2, anti HBsAg. 2 f12-h9;2 c4-b2;2C4-A2，抗HBsAg。

Hybridoma Pub Date : 2000-06-01 DOI: 10.1089/02724570050109693

引用次数: 0