Hybridoma and hybridomics最新文献_第9页

Anti-idiotypic antibody facilitates scFv chimeric immune receptor gene transduction and clonal expansion of human lymphocytes for tumor therapy. 抗独特型抗体促进scFv嵌合免疫受体基因转导和人淋巴细胞克隆扩增用于肿瘤治疗。

Hybridoma and hybridomics Pub Date : 2003-08-01 DOI: 10.1089/153685903322328938

Nai-Kong V Cheung, Hong-Fen Guo, Shakeel Modak, Irene Y Cheung

{"title":"Anti-idiotypic antibody facilitates scFv chimeric immune receptor gene transduction and clonal expansion of human lymphocytes for tumor therapy.","authors":"Nai-Kong V Cheung, Hong-Fen Guo, Shakeel Modak, Irene Y Cheung","doi":"10.1089/153685903322328938","DOIUrl":"https://doi.org/10.1089/153685903322328938","url":null,"abstract":"Chimeric immune receptors (CIR) transduced into lymphocytes link target recognition by single chain antibody Fv (scFv) to activation through CD28/TCRzeta signaling. As surrogate antigens, anti-idiotypic antibodies may facilitate gene-transduction and clonal expansion of human lymphocytes for in vivo tumor therapy. The murine monoclonal antibody (MAb) 8H9 reacts with a novel antigen widely expressed on solid tumors. A CIR consisting of human CD8-leader sequence, 8H9-scFv, CD28 (transmembrane and cytoplasmic domains), and TCR-zeta chain was constructed, ligated into the pMSCVneo vector, and used to transfect the packaging line GP + envAM12 bearing an amphotropic envelope. Rat anti-idiotypic MAb 2E9 (IgG2a) was used to clone retroviral producer line as well as to expand gene-modified primary human lymphocytes. Sequential enrichments using either affinity chromatography or cell sorting using anti-idiotypic MAb 2E9 significantly improved the percentage of producer clones positive for surface 8H9-scFv and the efficiency of their supernatant in transducing the indicator cell line K562. By 3 weeks of in vitro culture, >95% of transduced primary human lymphocytes were CIR-positive. Upon periodic stimulation with 2E9, these lymphocytes underwent >10(6)-fold expansion by 6 months in culture. They mediated antigen-specific non-MHC restricted cytokine release and tumor cytotoxicity, and inhibited human xenograft engraftment in SCID mice. Anti-idiotypic antibody may provide a useful tool for optimizing gene transduction of CIR fusion constructs into primary human lymphocytes and their continual expansion in vitro.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"22 4","pages":"209-18"},"PeriodicalIF":0.0,"publicationDate":"2003-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/153685903322328938","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40825462","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

Prediction of promiscuous and high-affinity mutated MHC binders. 混杂和高亲和力突变MHC结合物的预测。

Hybridoma and hybridomics Pub Date : 2003-08-01 DOI: 10.1089/153685903322328956

Manoj Bhasin, G P S Raghava

{"title":"Prediction of promiscuous and high-affinity mutated MHC binders.","authors":"Manoj Bhasin, G P S Raghava","doi":"10.1089/153685903322328956","DOIUrl":"https://doi.org/10.1089/153685903322328956","url":null,"abstract":"The identification of peptides in an antigenic sequence that can bind with high affinity to a wide range of MHC alleles is one of the challenges in subunit vaccine design. The mutation of natural peptides is an alternative to obtaining peptides that can bind to a wide range of MHC alleles with high affinity. A large number of experiments are typically necessary to identify mutations that define high-affinity binding peptides. Therefore there is a need to develop a computational method for detecting amino acid mutations in a peptide for making it high-affinity or promiscuous MHC binders. This report describes a high-throughput computer driven solution for the identification of promiscuous and high-affinity mutated binders of 47 MHC class I alleles by introducing mutations in an antigenic sequence. The method implements quantitative matrices for creating optimal mutations in an antigenic sequence. It has two major options: (i) prediction of promiscuous MHC binders and (ii) prediction of high-affinity binders. In case of prediction of promiscuous binders, the server allows a user to select (i) permissible mutations in a peptide; (ii) MHC alleles to whom it should bind; and (iii) positions at which mutation is allowed. In the case of prediction of high-affinity binders, the server allows users to specify the positions that should be conserved in the native protein. In both cases, the method computes the type of mutations and position of mutations in 9-mer peptides required to have the desired results. The web server MMBPred is available at www.imtech.res.in/raghava/mmbpred/.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"22 4","pages":"229-34"},"PeriodicalIF":0.0,"publicationDate":"2003-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/153685903322328956","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40825464","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: 43

Chimeric TNT-3 antibody/murine interferon-gamma fusion protein for the immunotherapy of solid malignancies. 嵌合的TNT-3抗体/小鼠干扰素- γ融合蛋白用于实体恶性肿瘤的免疫治疗。

Hybridoma and hybridomics Pub Date : 2003-08-01 DOI: 10.1089/153685903322328929

Myra M Mizokami, Peisheng Hu, Leslie A Khawli, Jiali Li, Alan L Epstein

{"title":"Chimeric TNT-3 antibody/murine interferon-gamma fusion protein for the immunotherapy of solid malignancies.","authors":"Myra M Mizokami, Peisheng Hu, Leslie A Khawli, Jiali Li, Alan L Epstein","doi":"10.1089/153685903322328929","DOIUrl":"https://doi.org/10.1089/153685903322328929","url":null,"abstract":"Interferon-gamma (IFN-gamma) has been used in the experimental treatment of cancer with limited success. Despite direct cytotoxic effects on tumor cells and the ability to stimulate the antitumor activities of a variety of effector cells, IFN-gamma has not been found to produce impressive therapeutic responses partly because of inadequate sustained intratumoral concentrations and systemic toxicity. To overcome these obstacles, we have developed an antibody/murine IFN-gamma fusion protein (chTNT-3/muIFN-gamma), which utilizes the tumor necrosis therapy antibody, chTNT-3, to target murine IFN-gamma to necrotic regions of solid tumors implanted in immunocompetent BALB/c mice. The genetically engineered fusion protein was expressed in NS0 cells using the Glutamine Synthetase Gene Amplification Expression System. After purification, the fusion protein demonstrated both antigen targeting and cytokine activities as assessed by in vitro assays which, when compared to recombinant free IFN-gamma, demonstrated approximately 40-45% biologic activity by two separate assay determinations. Pharmacokinetic and biodistribution studies in mice demonstrated a relatively long whole body half-life of 32 h in vivo and significant intratumoral accretion, respectively. Most importantly, immunotherapeutic studies in the MAD109 syngeneic murine carcinoma of the lung demonstrated significant intratumoral infiltration by leukocytes, primarily by macrophages and CD4(-) CD8(-) Thy-1.2(+) lymphocytes. Additionally, intravenous administration of the fusion protein significantly decreased the number of metastatic foci in an experimental model of pulmonary metastasis without causing any observable toxicity. These studies demonstrate that chTNT3/muIFN-gamma can safely target syngeneic tumor models as part of a promising strategy for the targeted immunotherapy of solid tumors.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"22 4","pages":"197-207"},"PeriodicalIF":0.0,"publicationDate":"2003-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/153685903322328929","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40825523","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: 32

Cassette vectors for conversion of Fab fragments into full-length human IgG1 monoclonal antibodies by expression in stably transformed insect cells. 在稳定转化的昆虫细胞中表达Fab片段转化为全长人IgG1单克隆抗体的盒式载体。

Hybridoma and hybridomics Pub Date : 2003-06-01 DOI: 10.1089/153685903322286548

Mary C Guttieri, Tanima Sinha, Carol Bookwalter, Mifang Liang, Connie S Schmaljohn

{"title":"Cassette vectors for conversion of Fab fragments into full-length human IgG1 monoclonal antibodies by expression in stably transformed insect cells.","authors":"Mary C Guttieri, Tanima Sinha, Carol Bookwalter, Mifang Liang, Connie S Schmaljohn","doi":"10.1089/153685903322286548","DOIUrl":"https://doi.org/10.1089/153685903322286548","url":null,"abstract":"Phage display technology allows for the production and rapid selection of antigen-specific, Fab antibody fragments. For purposes of immune therapy, though, complete antibodies that retain the Fc domain are often required. In this regard, we designed cassette vectors for converting human Fab fragments selected from combinatorial phage display libraries into full-length IgG(1) monoclonal antibodies (MAbs). Two expression vectors, pIEI-Light and pIEI-Heavy, were engineered to contain respective light- and heavy-chain human signal sequences downstream of the baculovirus immediate early gene promoter, IEI. Vector pIEI-Heavy also contains the coding region for each of the human IgG(1) constant domains. To generate complete antibody genes, the cassette vectors possess convenient restriction enzyme sites for rapid in-frame cloning of coding regions for full-length light chains in pIEI-Light and for the heavy-chain variable domains in pIEI-Heavy of Fab fragments. Using these constructs and a method that allows for stable transformation of insect cells, complete light- and heavy-chain genes can be inserted into the insect cell genome and subsequently expressed under the control of the baculovirus IEI promoter. This cassette vector system was used to generate stably transformed insect cells that continuously secreted functional full-length, IgG(1) MAbs. The expressed antibodies exhibited light and heavy chains of the appropriate molecular sizes and retained the ability to bind antigen. We conclude that our cassette vectors could serve as valuable tools for generating human IgG(1) antibodies.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"22 3","pages":"135-45"},"PeriodicalIF":0.0,"publicationDate":"2003-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/153685903322286548","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22559251","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: 4

Production of monoclonal antibody inhibiting dipeptidylaminopeptidase IV activity of Porphyromonas gingivalis. 抑制牙龈卟啉单胞菌二肽氨基肽酶IV活性单克隆抗体的制备。

Hybridoma and hybridomics Pub Date : 2003-06-01 DOI: 10.1089/153685903322286557

K Teshirogi, M Hayakawa, T Ikemi, Y Abiko

引用次数: 2

Partial characterization of the human serum transferrin epitope reactive with the monoclonal antibody TRC-2. 人血清转铁蛋白表位与单克隆抗体TRC-2反应的部分鉴定。

Hybridoma and hybridomics Pub Date : 2003-06-01 DOI: 10.1089/153685903322286584

S Oztürk, B Cirakoglu, E Bermek

{"title":"Partial characterization of the human serum transferrin epitope reactive with the monoclonal antibody TRC-2.","authors":"S Oztürk, B Cirakoglu, E Bermek","doi":"10.1089/153685903322286584","DOIUrl":"https://doi.org/10.1089/153685903322286584","url":null,"abstract":"A murine monoclonal antibody (MAb) (TRC-2) specific for human serum transferrin (Tf(h)) was developed. This antibody was depressive on cell growth in serum-free medium in the presence of limiting amounts of Tf(h), but it did not inhibit the binding of Tf(h)-alkaline phosphatase (AP) conjugate to the Tf-receptor (TfR) in a cellular enzyme-linked immunosorbent assay (CELISA) system. On the other hand, the immune complex Tf(h)-TRC-2 was implicated to bind to the receptor in indirect CELISA. Moreover, the detectability of Tf(h)-TfR on the cell surface via Tf-bound TRC-2 suggested that the antibody may inhibit the rapid internalization of this complex. To map the TRC-2-specific epitope, Tf(h) was subjected to proteolytic degradation following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. The treatment with trypsin gave rise to, among others, a fragment of about 42 kDa, which was reactive with TRC-2. Through sequence analysis by automated Edman degradation, the N-terminal sequence of the 42 kDa-tryptic fragment was aligned to the N-terminus of mature transferrin (VPDKTVR). The N-terminal sequence of an immunoreactive CNBr-fragment of about 13 kDa was, in turn, identical with the sequence (NQLRGKK) corresponding to the residues 110-116 on Tf(h).","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"22 3","pages":"165-71"},"PeriodicalIF":0.0,"publicationDate":"2003-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/153685903322286584","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22559255","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

Production of monoclonal antibody, PR81, recognizing the tandem repeat region of MUC1 mucin. 生产单克隆抗体PR81，识别MUC1粘蛋白串联重复区。

Hybridoma and hybridomics Pub Date : 2003-06-01 DOI: 10.1089/153685903322286566

M Paknejad, M J Rasaee, F Karami Tehrani, S Kashanian, M A Mohagheghi, K Omidfar, M Rajabi Bazl

{"title":"Production of monoclonal antibody, PR81, recognizing the tandem repeat region of MUC1 mucin.","authors":"M Paknejad, M J Rasaee, F Karami Tehrani, S Kashanian, M A Mohagheghi, K Omidfar, M Rajabi Bazl","doi":"10.1089/153685903322286566","DOIUrl":"https://doi.org/10.1089/153685903322286566","url":null,"abstract":"A monoclonal antibody (MAb) was generated by immunizing BALB/c mice with homogenized breast cancerous tissues. This antibody (PR81) was found to be of IgG(1) class and subclass, containing kappa light chain. PR81 reacted with either the membrane extracts of several breast cancerous tissues or the cell surface of some MUC1 positive cell lines (MCF-7, BT-20 and T-47D) tested by enzyme immunoassay and for MCF-7 by immunofluorescence method. PR81 also reacted with two synthetic 27 and 16-amino acid peptides, TSA-P1-24 and A-P1-15, respectively, which included the core tandem repeat sequence of MUC1. However, this antibody did not react with a synthetic 14 amino acid peptide that has no similarity with tandem repeat found in MUC1. The generated antibody had good and similar affinities (2.19 x 10(8) M(-1)) toward TSA-P1-24 and A-P1-15, which are mainly shared in the hydrophilic sequence of PDTRPAP. Through Western blot analysis of homogenized breast tissues, PR81 recognized only a major band of 250 kDa. This band is stronger in malignant tissue than benign and normal tissues.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"22 3","pages":"153-8"},"PeriodicalIF":0.0,"publicationDate":"2003-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/153685903322286566","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22559253","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: 26

Production and characterization of monoclonal antibodies against hepatitis B viruses and application of a quick sandwich ELISA. 抗乙型肝炎病毒单克隆抗体的制备、鉴定及快速夹心ELISA的应用

Hybridoma and hybridomics Pub Date : 2003-06-01 DOI: 10.1089/153685903322286593

Fatima Yücel, Aliihsan Manav, Aynur Başalp

{"title":"Production and characterization of monoclonal antibodies against hepatitis B viruses and application of a quick sandwich ELISA.","authors":"Fatima Yücel, Aliihsan Manav, Aynur Başalp","doi":"10.1089/153685903322286593","DOIUrl":"https://doi.org/10.1089/153685903322286593","url":null,"abstract":"Hepatitis B virus (HBV) infection is a major health problem worldwide. The diagnosis of acute and chronic hepatitis B infection is based on the detection of hepatitis B surface antigen (HBsAg). We report here the development of hybrid cell producing monoclonal antibodies (MAbs) specific for HBsAg using hybridoma technology. BALB/c mice were immunized with a mixture of HBsAg subtype \"ad\" and subtype \"ay.\" Spleen and lymph nodes were used as a source of high-titer antibody producing lymphocytes and removed and fused with myeloma cells of F0 origin separately. In the five fusion experiment, enzyme-linked immunosorbent assay (ELISA) tests showed that among 1594 hybridomas only 5 hybrids (9D12, 2B7, 4G5, 2G3, and 6E7) reacted with HBsAg. These MAbs were characterized for use in the development of diagnostic kits based on sandwich ELISA test system. The MAbs were conjugated with horseradish peroxidase (HRP) and used in the quick sandwich ELISA system. This system is a quite practical and time-saving test system when compared with common and commercial sandwich ELISA for diagnosis of hepatitis B surface antigen in human serum.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"22 3","pages":"173-7"},"PeriodicalIF":0.0,"publicationDate":"2003-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/153685903322286593","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22559256","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

A new epitope on swine CD5 molecule detected by monoclonal antibody 5F12/9. 单克隆抗体5F12/9在猪CD5分子上检测到一个新的表位。

Hybridoma and hybridomics Pub Date : 2003-06-01 DOI: 10.1089/153685903322286601

N Doménech, B Alvarez, R Bullido, F Alonso, A Ezquerra, J Domínguez

引用次数: 1

Monoclonal antibodies against blood group A secretors and nonsecretors saliva. 抗A型血分泌和非分泌唾液的单克隆抗体。

Hybridoma and hybridomics Pub Date : 2003-06-01 DOI: 10.1089/153685903322286610

Takeshi Ohmori, Hiroko Iwanari, Rie Aoi, Tomoko Shiraishi, Yukio Ito, Hajime Sato

引用次数: 2