Production, characterization and applications of mouse anti-grass carp (Ctenopharyngodon idellus) growth hormone monoclonal antibodies
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摘要Mouse anti-grass carp growth hormone (gcGH) monoclonal antibody (MAb) secretors were produced by PEG-mediated fusion of NS-1 myeloma cells and splenic B-lymphocytes of gcGH hyper-immunized mice. Positive secretors were screened by direct ELISA and cloned by limiting dilution. Three positive secretors, 21D3, 22G5 and 23B3, were obtained in a single fusion trial. Anti-gcGH MAbs were produced by growing hybridomas in the peritoneal cavity of pristane-primed mouse. The three MAbs were isotyped to be IgG2a, IgG2b and IgM, respectively. IgG MAbs were purified from ascitic fluid by Hitrap protein G column and IgM MAb was purified by gel filtration chromatography. The purified MAbs were highly specific and had moderate binding affinity. The MAbs were successfully used for the purification of native gcGH from mature grass carp pituitary extract by one-step immunoaffinity chromatography, for the quantification of gcGH by competitive sandwich ELISA, and for the probing of somatotropes in grass carp pituitary by immunohistochemistry. (c) 2005 Elsevier Inc. All rights reserved.
著者Leung MYK, Ho WKK
期刊名稱Comparative Biochemistry and Physiology - Part B: Biochemistry and Molecular Biology
出版年份2006
月份1
日期1
卷號143
期次1
出版社ELSEVIER SCIENCE INC
頁次107 - 115
國際標準期刊號1096-4959
語言英式英語
關鍵詞affinity chromatography; ELTSA; grass carp; growth hormone; hybridoma; immunohistochemistry; monoclonal antibody; somatotropes
Web of Science 學科類別Biochemistry & Molecular Biology; BIOCHEMISTRY & MOLECULAR BIOLOGY; Zoology; ZOOLOGY

上次更新時間 2021-11-05 於 01:10