標題: Distinction between dry and raw milk using monoclonal antibodies prepared against dry milk proteins
作者: Chen, WL
Huang, MT
Liu, HC
Li, CW
Mao, SJT
生物科技學系
Department of Biological Science and Technology
關鍵字: monoclonal antibody;dry milk;beta-lactoglobulin;thermal denaturation
公開日期: 1-八月-2004
摘要: It is well established that the heating process during the preparation of dry milk (DMLK) causes structural changes in some milk proteins. However, because such changes are subtle, whether they can be detected by an immunochemical approach remains questionable. The present study attempted to develop a sensitive mAb that might distinguish the DMLK from freshly prepared raw milk. To test this possibility, we immunized mice with commercially prepared DMLK and produced a panel of mAb. From 900 hybridomas screened using an ELISA, 4 clones were found to be specific to DMLK; the other 68 clones recognized both DMLK and raw milk. In contrast to polyclonal antibodies, only the specific mAb could detect the DMLK spiked into the raw milk at as low as 5% in concentration (vol/vol). Western blot analysis shows that these specific mAb were all directed against beta-lactoglobulin (LG) and LG-milk protein conjugates. These mAb reacted with raw milk heated at 95 for 15 min; the reaction with LG-conjugates, however, was abolished when treated with reducing reagent. Thus, results suggests that a new antigenic epitope was exposed in a heating process, and the thio group of LG cross linked with other protein moiety played a provocative role in mAb recognition. A hypothetical model with respect to the interaction between the mAb and DMLK is proposed and discussed.
URI: http://hdl.handle.net/11536/26491
ISSN: 0022-0302
期刊: JOURNAL OF DAIRY SCIENCE
Volume: 87
Issue: 8
起始頁: 2720
結束頁: 2729
顯示於類別:期刊論文