Hybridoma最新文献_第8页

Preparation and characterization of mabs against different epitopes of CD226 (PTA1). CD226 (PTA1)不同表位单抗的制备与鉴定

Hybridoma Pub Date : 2000-12-01 DOI: 10.1089/027245700750053986

W. Jia, Xue-song Liu, Yong Zhu, Qi Li, W. Han, Yun Zhang, Ji-Shuai Zhang, Kun Yang, Xin-hai Zhang, Boquan Jin

{"title":"Preparation and characterization of mabs against different epitopes of CD226 (PTA1).","authors":"W. Jia, Xue-song Liu, Yong Zhu, Qi Li, W. Han, Yun Zhang, Ji-Shuai Zhang, Kun Yang, Xin-hai Zhang, Boquan Jin","doi":"10.1089/027245700750053986","DOIUrl":"https://doi.org/10.1089/027245700750053986","url":null,"abstract":"Recently the platelet and T-cell activation antigen 1 (PTA1) was assigned as CD226 at the 7th Conference and Workshop on Human Leukocyte Differentiation antigens (HLDA). PTA1 is mainly expressed on activated T cells, natural killer (NK) cells, platelets and stimulated endotheliocytes, and involved in the differentiation of cytotoxic T lymphocytes (CTL) and NK, as well as platelet activation and aggregation. We raised hybridomas secreting monoclonal antibodies (MAbs) to PTA1 by using the natural PTA1 as immunogen, which was purified from platelets via affinity chromatography. These MAbs, designated FMU1, FMU2, FMU3, FMU4, FMU5, FMU6 and FMU7, could recognize PTA1 cDNA transfected COS7 cells detected by flow cytometry (FCM), and also react with both natural PTA1 and PTA1/Ig fusion protein in indirect enzyme-linked immunoadsorbent assay (ELISA). The biosensor epitope mapping assay showed that the seven MAbs, together with previous PTA1-specific MAbs Leo A1 and New E1, could bind seven distinct epitopes of PTA1, respectively. The panel of MAbs might be new powerful tools to study the structure-function relationship of PTA1 molecule, and to search for the ligand of PTA1.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"19 6 1","pages":"489-94"},"PeriodicalIF":0.0,"publicationDate":"2000-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245700750053986","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60496881","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: 17

Production and characterization of a new monoclonal antibody against Neisseria meningitidis: study of the cross-reactivity with different bacterial genera. 一种新的抗脑膜炎奈瑟菌单克隆抗体的制备与鉴定:与不同属细菌的交叉反应性研究。

Hybridoma Pub Date : 2000-12-01 DOI: 10.1089/027245700750053931

E. Gaspari

{"title":"Production and characterization of a new monoclonal antibody against Neisseria meningitidis: study of the cross-reactivity with different bacterial genera.","authors":"E. Gaspari","doi":"10.1089/027245700750053931","DOIUrl":"https://doi.org/10.1089/027245700750053931","url":null,"abstract":"We have generated a hybridoma cell line which produces an 8C7Br1 clone of the IgM antibody isotype. It recognizes the 50-, 65-, and 60-kDa antigens and is reactive with strains of N. meningitidis in the 98% of local Neisseria genera by Dot-ELISA assays. Two percent of the strains of N. meningitidis B do not present reactivity with the 8C7Br1 monoclonal antibody (MAb). The antibody reacted against N. meningitidis of serogroups A, B, C, X, Y, Z, and different serotypes and subtypes of N. meningitidis B and C by means of Dot-ELISA and Immunoblot. It cross-reacted with Neisseria gonorrhoeae, Neisseria lactamica, Haemophilus influenzae type b, Escherichia coli, Salmonella typhimurium, Salmonella typhi, Shigella flexneri, Bordetella pertussis, and Bacillus subtilis. The 8C7Br1 MAb reacted with the 65-kDa protein present in the prototype meningococcal strains B:16:B6(B2a:P1.5.2) and 2996 (B2b:P1.5.2). In H. influenzae type b, E. coli and B. subtilis, the MAb recognized the protein of 60, 65, and 70 kDa, respectively. FACS analysis showed that 8C7Brl MAb could recognize the 50-kDa protein on the surface of N. meningitidis homologous (B:4:P1.9) strain. These results, together with the bactericidal activity of 8C7Br1, and an experiment of passive protection in mice, demonstrated the potential importance of the cross-reactive protein as a candidate antigen for N. meningitidis B vaccine composition.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"19 6 1","pages":"445-53"},"PeriodicalIF":0.0,"publicationDate":"2000-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245700750053931","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60497055","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: 14

Production and characterization of an estrogen receptor beta subtype-specific mouse monoclonal antibody. 雌激素受体β亚型特异性小鼠单克隆抗体的制备和鉴定。

Hybridoma Pub Date : 2000-12-01 DOI: 10.1089/027245700750053977

J. Su, D. D. Mckee, B. Ellis, S. Kadwell, G. Wisely, L. Moore, J. Triantafillou, T. Kost, S. Fuqua, J. Moore

{"title":"Production and characterization of an estrogen receptor beta subtype-specific mouse monoclonal antibody.","authors":"J. Su, D. D. Mckee, B. Ellis, S. Kadwell, G. Wisely, L. Moore, J. Triantafillou, T. Kost, S. Fuqua, J. Moore","doi":"10.1089/027245700750053977","DOIUrl":"https://doi.org/10.1089/027245700750053977","url":null,"abstract":"An important step in differentiating the unique physiological roles of the alpha and beta forms of estrogen receptor is to determine the precise expression pattern of each of these receptors. We report the generation and characterization of a murine IgG1 monoclonal antibody (MAb), ER15.64A that is ERbeta subtype-specific and capable of recognizing full-length human ERbeta as well as all of its known protein isoforms. ER15.64A, raised against a ERbeta peptide (aa2-18)-keyhole limpet hemocyanine conjugate, reacted to the immunizing peptide and the full-length E. coli expressed ERbeta in ELISA and BIAcore assays. It also immunostained nuclei of Sf9 insect cells that were infected with an ERbeta-baculovirus. In Western analysis, ER15.64A recognized ERbeta1 and ERbeta2 proteins from a reticulocyte in vitro transcription/translation preparation. This antibody did not cross-react with recombinant ERalpha in ELISA, BIAcore, immunocytochemistry, or Western blot analysis. The specificity of ER15.64A should make this antibody a useful tool for monitoring expression of ERbeta and its isoforms at the protein level and should aid in distinguishing the pattern of ERbeta receptor expression from that of ERalpha.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"19 6 1","pages":"481-7"},"PeriodicalIF":0.0,"publicationDate":"2000-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245700750053977","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60496822","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

Immune response to 17beta-estradiol involved in polymer gels: antigen specificity and affinity of hybridoma clones. 聚合物凝胶对17 -雌二醇的免疫反应:杂交瘤克隆的抗原特异性和亲和力。

Hybridoma Pub Date : 2000-12-01 DOI: 10.1089/027245700750053995

A. Başalp, Z. Mustafaeva, M. Mustafaev, E. Bermek

引用次数: 10

Removal of amphipathic epitopes from genetically engineered antibodies: production of modified immunoglobulins with reduced immunogenicity. 从基因工程抗体中去除两亲性表位:产生免疫原性降低的修饰免疫球蛋白。

Hybridoma Pub Date : 2000-12-01 DOI: 10.1089/027245700750053959

C. Mateo, J. Lombardero, E. Moreno, A. Morales, G. Bombino, J. Coloma, L. Wims, S. Morrison, R. Pérez

{"title":"Removal of amphipathic epitopes from genetically engineered antibodies: production of modified immunoglobulins with reduced immunogenicity.","authors":"C. Mateo, J. Lombardero, E. Moreno, A. Morales, G. Bombino, J. Coloma, L. Wims, S. Morrison, R. Pérez","doi":"10.1089/027245700750053959","DOIUrl":"https://doi.org/10.1089/027245700750053959","url":null,"abstract":"Several approaches have been developed to reduce the human immune response to nonhuman antibodies. However, chimeric antibodies and humanized antibodies often have decreased binding affinity. We described a new approach for reducing the immunogenicity of chimeric antibodies while maintaining the affinity. This approach seeks to prevent the recognition of murine immunogenic peptides from the antibody variable region by human lymphocytes. Putative immunogenic epitopes in the variable region are identified and subjected to site directed mutagenesis to make them human and/or to break the amphipathic motifs. The R3 antibody, which blocks the epidermal growth factor (EGF) receptor, was used as a model system to test this approach. Four segments containing possible amphipathic epitopes were found in the heavy variable domain using the program AMPHI. Six amino acids within two of these segments were substituted by the corresponding residues from a homologous human sequence. No mutations were made in the murine light variable domain. Experiments in monkeys suggested that the \"detope\" R3 antibody was less immunogenic than its chimeric analogue. A search for possible amphipathic epitopes in the Kabat database revealed the presence of conserved patterns in the different families of variable region sequences, suggesting that the proposed method may be of general applicability.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"19 6 1","pages":"463-71"},"PeriodicalIF":0.0,"publicationDate":"2000-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245700750053959","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60497139","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: 42

Production and characterization of monoclonal antibodies directed to the kringle V and protease domains of human apolipoprotein(a). 针对人载脂蛋白(a) kringle V和蛋白酶结构域的单克隆抗体的制备和鉴定

Hybridoma Pub Date : 2000-12-01 DOI: 10.1089/027245700750053922

Y. Seo, K. You, J. Kwak

{"title":"Production and characterization of monoclonal antibodies directed to the kringle V and protease domains of human apolipoprotein(a).","authors":"Y. Seo, K. You, J. Kwak","doi":"10.1089/027245700750053922","DOIUrl":"https://doi.org/10.1089/027245700750053922","url":null,"abstract":"Production and use of anti-apolipoprotein(a) monoclonal antibodies (MAbs) specific to single copy regions in the polymorphic lipoprotein(a) (Lp(a)) has been emphasized to be important for the standardization of measurements of the coronary heart disease risk factor, Lp(a). Here, mouse MAbs were prepared against the kringle V (V) and protease (P) domains of human apolipoprotein(a) (apo(a)), which domains are present in single copy in the apo(a) molecule. The cDNA for apo(a)VP was cloned from human liver cDNA library, and the V-P recombinant protein overexpressed in Escherichia coli was used as an antigen for the antibody production. Two antibodies named as MAb(a)20 and MAb(a)23 were finally produced, and they were characterized for their binding specificity and epitopes. The specificity of the antibodies was confirmed by an immunoblotting procedure and an enzyme-linked immunoassay (ELISA). It was shown that the antibodies had little, if any, cross-reactivity with human plasminogen, which is relatively abundant in human serum and is highly homologous (85%) with apo(a) in amino acid (aa) sequence. For epitope analysis, 3'-deletional series of apo(a)VP cDNA were constructed, and expression products of them were analyzed for the binding MAb(a)20 and MAb(a)23 do. It has been revealed that distinct epitopes were recognized by the two MAbs: MAb(a)23 (gamma2b, kappa) bound to the V region about 60 aa downstream from the N-terminal, and MAb(a)20 (gamma1, kappa) bound to the P region close to the C-terminal. A one step-sandwich ELISA system for Lp(a) was developed using MAb(a)20 as a capturing antibody and horseradish peroxidase (HRP)-coupled MAb(a)23 as a detecting antibody. The assay was found to be sensitive and useful for detecting Lp(a) in the range of 4-150 microg/dL (80 pM-3 nM).","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"19 6 1","pages":"435-44"},"PeriodicalIF":0.0,"publicationDate":"2000-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245700750053922","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60496835","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: 2

Characterization of the in vitro and in vivo activity of monoclonal antibodies to human IL-18. 人IL-18单克隆抗体体外和体内活性的研究。

Hybridoma Pub Date : 2000-10-01 DOI: 10.1089/02724570050198875

S Holmes, J A Abrahamson, N Al-Mahdi, S S Abdel-Meguid, Y S Ho

引用次数: 16

Development and characterization of monoclonal antibodies to chicken riboflavin carrier protein. 鸡核黄素载体蛋白单克隆抗体的制备与鉴定。

Hybridoma Pub Date : 2000-10-01 DOI: 10.1089/02724570050198901

A Deshmukh, M Gani, U Natraj

引用次数: 0

Generation of monoclonal antibodies to cryptic collagen sites by using subtractive immunization. 利用减法免疫产生针对隐性胶原位点的单克隆抗体。

Hybridoma Pub Date : 2000-10-01 DOI: 10.1089/02724570050198893

J Xu, D Rodriguez, J J Kim, P C Brooks

{"title":"Generation of monoclonal antibodies to cryptic collagen sites by using subtractive immunization.","authors":"J Xu, D Rodriguez, J J Kim, P C Brooks","doi":"10.1089/02724570050198893","DOIUrl":"https://doi.org/10.1089/02724570050198893","url":null,"abstract":"The extracellular matrix (ECM) plays a fundamental role in the regulation of normal and pathological processes. The most abundantly expressed component found in the ECM is collagen. Triple helical collagen is known to be highly resistant to proteolytic cleavage except by members of the matrix metalloproteinase (MMP) family of enzymes. To date little is known concerning the biochemical consequences of collagen metabolism on human diseases. This is due in part to the lack of specific reagents that can distinguish between proteolyzed and triple helical forms of collagen. Here we used the technique of Subtractive Immunization (SI) to generate two unique monoclonal antibodies (MAbs HUIV26 and HUI77) that react with denatured and proteolyzed forms of collagen, but show little if any reaction with triple helical collagen. Importantly, HUIV26 and HUI77 react with cryptic sites within the ECM of human melanoma tumors, demonstrating their utility for immunohistochemical analysis in vivo. Thus, the generation of these novel MAbs not only identify specific cryptic epitopes within triple helical collagen, but also provide important new reagents for studying the roles of collagen remodeling in normal as well as pathological processes.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"19 5","pages":"375-85"},"PeriodicalIF":0.0,"publicationDate":"2000-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/02724570050198893","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21951315","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: 56

Generation and characterization of a neutralizing monoclonal antibody against erythroid cell stimulating factor. 抗红细胞刺激因子的中和性单克隆抗体的制备和鉴定。

Hybridoma Pub Date : 2000-10-01 DOI: 10.1089/02724570050198866

S Ponnappan, U Ponnappan, K B Udupa

{"title":"Generation and characterization of a neutralizing monoclonal antibody against erythroid cell stimulating factor.","authors":"S Ponnappan, U Ponnappan, K B Udupa","doi":"10.1089/02724570050198866","DOIUrl":"https://doi.org/10.1089/02724570050198866","url":null,"abstract":"Erythroid cell stimulating factor (ESF) is present in mouse serum and has been reported to function in concert with erythropoietin (EPO) in the formation of erythroid cells in in vitro culture systems. We report here the generation and characterization of a monoclonal antibody (MAb) directed against ESF, with potent anti-ESF-neutralizing activity. A hybridoma-producing MAb to ESF was selected following enzyme-linked immunosorbent assay (ELISA)-based screening of 270 colonies obtained from a fusion of immunized mouse splenocytes with NS1 myeloma cells. Western blot analyses of mouse serum using this antibody specifically detected a single protein (approximate molecular weight of 60 kDa and 120 kDa, under reducing and nonreducing conditions, respectively) corresponding to ESF, with no reactivity to EPO. Furthermore, this MAb demonstrated reactivity to a protein similar in molecular mass, across species, showing reactivity in sera obtained from human, horse, goat, guinea pig, rabbit, and rat. Immuno-chemical characterization demonstrated this antibody to be of IgG3 isotype, bearing kappa light chains. Injection of this monoclonal anti-ESF antibody to exhypoxic polycythemic mice at 6 and 24 h after EPO injection significantly reduced 59Fe incorporation into red blood cells, demonstrating its ability to neutralize in vivo erythropoiesis in our mouse model system. Thus, this novel erythroid cell-specific MAb will be an invaluable tool for further delineating the physiological role of ESF in in vivo erythropoiesis.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"19 5","pages":"355-61"},"PeriodicalIF":0.0,"publicationDate":"2000-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/02724570050198866","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21951312","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