Hybridoma最新文献_第4页

Monoclonal antibodies to Kaiso: a novel transcription factor and p120ctn-binding protein. Kaiso单克隆抗体:一种新的转录因子和p120ctn结合蛋白。

Hybridoma Pub Date : 2001-06-01 DOI: 10.1089/027245701750293484

J. Daniel, R. Ireton, A. Reynolds

引用次数: 22

Recombinant human interleukin-6 enhances the immunoglobulin secretion of a rabbit-rabbit hybridoma. 重组人白细胞介素-6增强兔-兔杂交瘤免疫球蛋白分泌。

Hybridoma Pub Date : 2001-06-01 DOI: 10.1089/027245701750293529

M. Liguori, J. Hoff-Velk, D. Ostrow

{"title":"Recombinant human interleukin-6 enhances the immunoglobulin secretion of a rabbit-rabbit hybridoma.","authors":"M. Liguori, J. Hoff-Velk, D. Ostrow","doi":"10.1089/027245701750293529","DOIUrl":"https://doi.org/10.1089/027245701750293529","url":null,"abstract":"The generation of stable rabbit-rabbit hybridomas is now possible by the recent development of a rabbit fusion partner. The ability to generate rabbit monoclonal antibodies (MAbs) can be advantageous because these rabbit immunoglobulins tend to exhibit higher affinity than murine MAbs. Furthermore, it has been observed that, in general, rabbits will elicit an immune response to antigens of limited immunogenicity in mice. Unfortunately, these rabbit-rabbit hybridomas secrete only 200 ng/mL to 5 microg/mL of immunoglobulin, which may limit larger scale production of rabbit antibodies. This study sought to determine if interleukin 6 (IL-6), which has been reported to have proliferative and secretory stimulating effects on some murine hybridomas, had any effect on a rabbit cell line that secretes a monoclonal IgG specific for estradiol. The results demonstrated that recombinant human IL-6 had a dose-dependent enhancing effect on the IgG secretion of the rabbit-rabbit hybridoma. The enhancing effect was consistent when the cells were continuously passed in the presence of IL-6. However, IL-6 did not affect the growth of the hybridoma. In contrast, no discernible effect was accomplished with recombinant mouse IL-6. Furthermore, no basal IL-6 activity was detected in the rabbit hybridoma extracellular medium. The IL-6 enhancement effect observed in this study may help to increase the immunoglobulin yield of rabbit hybridomas and to assist in the understanding of the mechanism(s) behind the lowered secretion level.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 3 1","pages":"189-98"},"PeriodicalIF":0.0,"publicationDate":"2001-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245701750293529","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60500067","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: 18

Detection of superficial zone protein in human and animal body fluids by cross-species monoclonal antibodies specific to superficial zone protein. 浅表带蛋白特异性跨种单克隆抗体检测人和动物体液中浅表带蛋白。

Hybridoma Pub Date : 2001-06-01 DOI: 10.1089/027245701750293475

J. Su, B. Schumacher, K. M. Lindley, V. Soloveychik, W. Burkhart, J. Triantafillou, K. Kuettner, T. Schmid

{"title":"Detection of superficial zone protein in human and animal body fluids by cross-species monoclonal antibodies specific to superficial zone protein.","authors":"J. Su, B. Schumacher, K. M. Lindley, V. Soloveychik, W. Burkhart, J. Triantafillou, K. Kuettner, T. Schmid","doi":"10.1089/027245701750293475","DOIUrl":"https://doi.org/10.1089/027245701750293475","url":null,"abstract":"In this report we describe the purification of human superficial zone protein (SZP), the generation of cross-species monoclonal antibodies (MAbs) and the detection of this protein in human and animal body fluids. Human SZPs, used as immunizing antigens, were purified either from culture media of human cartilage organ cultures or from human synovial fluids. The immunizing antigens were mixed with RIBI adjuvant in one of three forms: nonmodified SZP, superficial zone protein-keyhole limpet hemocyanin conjugate (SZP-KLH), or a mixture of superficial zone protein and hyaluronic acid (SZP-HA). A panel of MAbs including GW4.23, S6.79, S13.52, S13.233, and S17.109 were generated and characterized. Monoclonal antibody (MAb) S6.79, an IgG2b with K(D) 3.14 x 10(-9) M from SZP-KLH immunization, is of particular interest. It reacts strongly to a large molecular weight form of SZP in both enzyme-linked immunosorbent assay (ELISA) and Western blotting. It stains the most superficial layer of articular cartilage in immunohistochemistry, whereas the middle and deep zones of cartilage are not stained. When MAb S6.79 was applied to Western blots of human body fluids, a strong 345-kDa band was detected in samples of synovial fluid and weaker bands of similar size were detected in samples of plasma and serum. MAb S6.79 also showed cross-species immunoreactivity with SZP in samples of synovial fluids harvested from bovine, dog, guinea pig, and rabbit, as demonstrated by Western blotting and antibody absorption experiments. This cross-species MAb will be a useful tool in human and animal model studies for monitoring SZP levels and tissue distribution. It may help define the roles of SZP in normal articular joints and may be of diagnostic or prognostic value for the measurement of SZP in pathological conditions such as osteoarthritis, rheumatoid arthritis, and camptodactyly-arthropathy-coxa vara-pericarditis.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 3 1","pages":"149-57"},"PeriodicalIF":0.0,"publicationDate":"2001-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245701750293475","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60499396","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: 88

Generation and characterization of recombinant ScFv antibodies detecting Eimeria acervulina surface antigens. 检测尖角艾美耳球虫表面抗原的重组ScFv抗体的制备与鉴定。

Hybridoma Pub Date : 2001-06-01 DOI: 10.1089/027245701750293501

J. K. Kim, W. Min, H. Lillehoj, S. Kim, E. Sohn, K. Song, J. Y. Han

{"title":"Generation and characterization of recombinant ScFv antibodies detecting Eimeria acervulina surface antigens.","authors":"J. K. Kim, W. Min, H. Lillehoj, S. Kim, E. Sohn, K. Song, J. Y. Han","doi":"10.1089/027245701750293501","DOIUrl":"https://doi.org/10.1089/027245701750293501","url":null,"abstract":"In our previous attempt to generate monoclonal antibodies (MAbs) against coccidia parasites that more accurately reflect the natural avian humoral immune response, we produced two chicken B-cell hybridomas, 5D11 and 2-1. While both cell lines secreted antibodies reactive with sporozoites of Eimeria acervulina, they were produced in yields too low to conduct meaningful in vivo studies. To circumvent this problem, we produced four single chain variable fragment (scFv) antibodies from the V(H) and V(L) genes of hybridomas 5D11 and 2-1. The concentration of these recombinant antibodies expressed in E. coli and purified to homogeneity was 5-6 mg/L. Three of the antibodies exhibited antigen binding specificity to Eimeria surface antigens equivalent to that of the native MAbs. Nucleotide sequence analysis of the V(L) genes from hybridomas 5D11 and 2-1 and genomic DNA revealed vestiges of gene conversion with V(lambda) pseudogenes. These recombinant scFv antibodies will prove useful for further characterization of natural Eimeria surface antigens as potential vaccine candidates.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 3 1","pages":"175-81"},"PeriodicalIF":0.0,"publicationDate":"2001-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245701750293501","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60499928","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: 15

Preparation of monoclonal antibodies against human telomerase. 人端粒酶单克隆抗体的制备。

Hybridoma Pub Date : 2001-06-01 DOI: 10.1089/027245701750293510

H. Kaur, E. Richardson, L. Murty

{"title":"Preparation of monoclonal antibodies against human telomerase.","authors":"H. Kaur, E. Richardson, L. Murty","doi":"10.1089/027245701750293510","DOIUrl":"https://doi.org/10.1089/027245701750293510","url":null,"abstract":"Telomerase, a ribonucleoprotein enzyme that extends telomeres of eukaryotic chromosomes, consists of the catalytic protein submit telomerase reverse transcriptase (TERT) and a telomerase RNA subunit. Nearly 85% of human tumors have tested positive for high telomerase activity. Telomerase activity is very low or not present in normal cells, whereas it is up-regulated in immortalized cells. Telomerase, partially purified from the breast tumor cell line MCF-7, was used to immunize Balb/C mice. Monoclonal antibodies (MAbs) were prepared by conventional hybridoma technology and screened by enzyme-linked immunoadsorbent assay (ELISA), followed by a polymerase chain reaction (PCR) based telomeric amplification repeat protocol (TRAP) assay to detect binding to or inhibition of telomerase activity. Reactive MAbs were found to be of IgM type by mu specific ELISA. Two MAbs were characterized, one that neutralizes telomerase activity in TRAP assay and the other non-neutralizing. In Western blotting, crude telomerase extract and HIV-1 virus lysate (control) were blotted on nitrocellulose membranes and the strips were treated with both MAbs and a nonrelated HIV polymerase-specific MAb, also IgM type. A band of approx. 65-kDa was detected in extracts of 293 cells with both the MAbs, but no reaction occurred with the HIV polymerase-specific MAb used as control. Similarly, when HIV-1 virus lysate strips were treated with HIV polymerase-specific MAb, a 65-kDa band was detected and no band was observed with either of the hybridoma supernatants. These antibodies may be useful for studying regulatory mechanism of telomerase and inhibition of its activity in vitro and in vivo.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 3 1","pages":"183-8"},"PeriodicalIF":0.0,"publicationDate":"2001-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245701750293510","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60500006","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

Characterization of monoclonal antibodies with specificity for the core oligosaccharide of Shigella lipopolysaccharide. 志贺氏菌脂多糖核心寡糖特异性单克隆抗体的鉴定。

Hybridoma Pub Date : 2001-04-01 DOI: 10.1089/02724570152057571

S. R. Rahman, W. H. Stimson

引用次数: 5

Phase I clinical evaluation of a neutralizing monoclonal antibody against epidermal growth factor receptor in advanced brain tumor patients: preliminary study. 抗表皮生长因子受体中和性单克隆抗体在晚期脑肿瘤患者中的I期临床评价:初步研究。

Hybridoma Pub Date : 2001-04-01 DOI: 10.1089/02724570152057634

T. Crombet, O. Torres, V. Rodríguez, A. Menendez, A. Stevenson, M. Ramos, F. Torres, R. Figueredo, I. Veitia, N. Iznaga, R. Pérez, A. Lage

{"title":"Phase I clinical evaluation of a neutralizing monoclonal antibody against epidermal growth factor receptor in advanced brain tumor patients: preliminary study.","authors":"T. Crombet, O. Torres, V. Rodríguez, A. Menendez, A. Stevenson, M. Ramos, F. Torres, R. Figueredo, I. Veitia, N. Iznaga, R. Pérez, A. Lage","doi":"10.1089/02724570152057634","DOIUrl":"https://doi.org/10.1089/02724570152057634","url":null,"abstract":"High levels of growth factors and their receptors have been demonstrated in human tumors. Gliomas and meningiomas are characterized by overexpression of epidermal growth factor receptor (EGF-R). Ior egf/r3, is a neutralizing murine monoclonal antibody (MAb) against EGF-R, and was generated at the Cuban Institute of Oncology. The antibody recognizes EGF-R with high affinity, inhibiting tyrosine kinase activation. A clinical trial was conducted in brain tumor patients to evaluate toxicity, immunogenicity, and clinical benefit of escalating doses of the antibody. Nine patients with histologically confirmed gliomas or meningiomas, who had active or recurrent disease after receiving conventional treatment, received four intravenous doses of ior egf/r3. Total dosages ranged from 160 to 480 mg. As inclusion criteria, radioimmunoscintigraphy with the same MAb labeled with 99mTechnetium (99mTc) was performed. Immune response against the murine antibody was also evaluated. After four doses of ior egf/r3 MAb, no significant toxicity was found, except in one patient who developed a grade 4 allergic adverse event. This reaction was probably related with previous sensitization to the same MAb and the development of human anti-mouse antibodies (HAMA) response. Despite no major objective antitumor responses, eight patients had stable disease on the 6-month evaluation, and two patients remain alive after four years of MAb therapy.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 2 1","pages":"131-6"},"PeriodicalIF":0.0,"publicationDate":"2001-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/02724570152057634","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60498809","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: 61

Characterization of four new monoclonal antibodies that recognize mouse natural killer activation receptors. 四种识别小鼠自然杀伤激活受体的单克隆抗体的鉴定。

Hybridoma Pub Date : 2001-04-01 DOI: 10.1089/02724570152057580

S. Kung, R. Su, J. Shannon, R. Miller

{"title":"Characterization of four new monoclonal antibodies that recognize mouse natural killer activation receptors.","authors":"S. Kung, R. Su, J. Shannon, R. Miller","doi":"10.1089/02724570152057580","DOIUrl":"https://doi.org/10.1089/02724570152057580","url":null,"abstract":"With the aim of identifying natural killer (NK) activation receptors, we immunized BALB/c mice with (BALB/cxB6)F1 NK LAK cells and made B-cell hybridomas. These were screened for monoclonal antibody (MAb) reacting with an NK activation receptor by using an antibody-induced redirected lysis (AIRL) assay against FcR-bearing P815 targets. Four hybridomas, clones 1C10, 1F10, 2D10 and 4G4, were selected for further characterization. Protein G-purified MAbs from these clones activated both resting and IL-2 activated B6 or F1 NK cells in the AIRL assay. 1F10 MAb, but not the other three MAbs, could compete for the binding of anti-NK1.1 (PK136) MAb to F1 NK cells. The four MAbs were screened for their ability to bind to or activate NK cells from the mouse strains SJL/J, DBA/2, 129/J, C3H/J, and BALB.K. None showed activity except IC10, which could bind to and activate SJL/J NK cells. When members of the NKR-P1 family from both B6 mice (A, B, and C genes expressed) and SJL mice (only A and B genes expressed) were expressed in Jurkat cells and tested for their antibody reactivity, PK136 MAb was found to recognize B6 NKR-P1C and SJL/J NKR-P1B; IC10 MAb was found to recognize NKR-P1-A, -B and -C from B6, but not NKR-P1A or -B from SJL/J; and 1F10 MAb was found to react only with B6 NKR-P1C.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 2 1","pages":"91-101"},"PeriodicalIF":0.0,"publicationDate":"2001-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/02724570152057580","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60498794","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

Characterization of monoclonal antibodies specific for feline serum amyloid (SAA) protein. 猫血清淀粉样蛋白(SAA)特异性单克隆抗体的鉴定。

Hybridoma Pub Date : 2001-04-01 DOI: 10.1089/02724570152057599

K. Sasaki, Z. Ma, K. Okazaki, T. Khatlani, M. Okuda, T. Kajikawa, T. Onishi

{"title":"Characterization of monoclonal antibodies specific for feline serum amyloid (SAA) protein.","authors":"K. Sasaki, Z. Ma, K. Okazaki, T. Khatlani, M. Okuda, T. Kajikawa, T. Onishi","doi":"10.1089/02724570152057599","DOIUrl":"https://doi.org/10.1089/02724570152057599","url":null,"abstract":"Serum amyloid A (SAA) has been characterized as an inflammatory marker in many species. In this study, we have developed and characterized monoclonal antibodies (MAbs) against feline SAA (fSAA) derived from culture hybridomas. These hybridomas were produced from the fusion of Balb/c-derived myeloma s/p20-Ag14 and splenocytes from mice immunized with purified recombinant feline SAA (rfSAA). Six hybridomas secreting MAbs, M2, M5, M7, M8, M13, and M15, were selected and subcloned on the basis of their specificity to rfSAA by enzyme-linked immunoabsorbent assay (ELISA), and confirmed based on their specificity to rf-SAA by immunoblot analysis. Out of six clones, two clones (M5 and M7) showed higher reactivity with rf-SAA, and were selected for further analysis of ELISA additivity and Western blot cross-reactivity tests. As a result, M5 and M7 clones recognized the same or excessively near epitopes on rfSAA and reacted with rfSAA, fSAA and equine recombinant SAA, but showed no reaction with human recombinant SAA. Because of their specificity, these MAbs may be usefully applied in studying the measurement of SAA concentration in cat serum.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 2 1","pages":"103-8"},"PeriodicalIF":0.0,"publicationDate":"2001-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/02724570152057599","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60498849","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

Monoclonal antibody to rat galanin: production, characterization, and in vivo immunoneutralization activity. 大鼠丙氨酸单克隆抗体:生产、鉴定和体内免疫中和活性。

Hybridoma Pub Date : 2001-04-01 DOI: 10.1089/02724570152057607

H. Wong, C. Sternini, H. Yang, T. Pham, J. Walsh

{"title":"Monoclonal antibody to rat galanin: production, characterization, and in vivo immunoneutralization activity.","authors":"H. Wong, C. Sternini, H. Yang, T. Pham, J. Walsh","doi":"10.1089/02724570152057607","DOIUrl":"https://doi.org/10.1089/02724570152057607","url":null,"abstract":"A monoclonal antibody (MAb) to galanin was prepared by cell fusion of myeloma Fox-NY and spleen cells from Robertsonian mice immunized with rat galanin. Hybridomas producing high-affinity antibodies were cloned in pristine-primed Balb/c mice. The antibody was purified by affinity chromatography and concentrated to 12 mg IgG/mL by dialysis. Immunoreactivity of the antibody was screened by radioimmunoassay. Ascites fluid contained approximately 10 mg/mL IgG that belong to the subclass of IgG2a as determined by enzyme-linked immunoadsorbent assay (ELISA). The titer of this IgG2a antibody entitled #G65G was 1:10,000 and the ID50 for rat galanin was 1000 fmol/mL as determined by liquid phase radioimmunoassay. Immunohistochemistry showed that this galanin MAb stains densed, beaded processes distributed to the enteric plexuses, where they appear to encircle neuronal cell bodies, to the muscle layer, where they are particularly abundant in the circular muscle layer and in the deep muscular layer, and to the mucosa. In vivo capacity of immunoneutralization by this antibody was tested in male Sprague-Dawley rats fasted for 24 h and anesthetized with urethane. Systemic injection of protein A purified galanin antibody (6 mg/rat) decreased by 70% of the inhibitory effect of intravenous galanin (2 nmol/kg/h i.v.) on gastric acid secretion induced by intracisternal TRH analog. These results show that galanin antibody #G65G is useful for in vivo immunoneutralization of galanin effects and is a valuable tool for immunohistochemical localization of galanin in gastrointestinal tissues.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 2 1","pages":"109-15"},"PeriodicalIF":0.0,"publicationDate":"2001-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/02724570152057607","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60498931","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