Mouse Monoclonal Antibodies against Bovine Milk Lipoprotein Lipase: Production and Characterization
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    Abstract:

    Aim Studies were performed to produce specificmonoclonal antibodies to bovine milk lipoprotein lipase(bLPL), to verify the specificity of the monoclonal an-tibodies for bLPL, and to characterize them.Metkods Purified lipoprotein lipase (LPL) frombovine milk-56 kDa was used as an antigen for the pro-duction of anti-LPL antibodies in mice. The spleen wasremoved from the animal having the highest titer of an-tibodies to LPL and the cells were fused with mousemyeloma cells. After cultured with hypoxanthine-aminopterin-thymidine (HAT ) culture medium, thisprocedure thus identified only those hybridomas whichproduced antibodies directed against LPL for super-natants by enzyme linked immunosorbent assay(ELISA). After cloning five monoclonal cell strainswere established, named as 2E5, 2G10, 6D7, 8D2 and7F4. The titriton of monoclonal antibodies (McAb) are5 × 10-2~2 × 10-3 for supernatants by ELISA.Results and Conclusion Contitration study showedMcAb recognized three different epitopes of LPL: 6D7and 8D2 to a close epitop; 2G10 and 7F4 to other closeepitope; 2G10 but recognized a far epitope away fromthe two of others. Western blot analysis of bovine milkwith the McAb gave a single protein band of 56 kDa;2G10 gave two protein bands of 50 kDa and 47 kDa.Dot-blot analysis of bovine milk with the McAb 2E5,8D2 and 6D7 showed specific reaction Western blot andDet-blot analysis of human milk with the McAb, hadno cross-reacted.

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HOU Zhe; LIU De-Wen; WO Xing-De, ZHAO Ge-Ping. Mouse Monoclonal Antibodies against Bovine Milk Lipoprotein Lipase: Production and Characterization[J]. Editorial Office of Chinese Journal of Arteriosclerosis,1997,5(2):116-120.

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History
  • Received:March 17,1997
  • Revised:June 13,1997
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