標題: A novel conformation-dependent monoclonal antibody specific to the native structure of beta-lactoglobulin and its application
作者: Chen, WL
Liu, WT
Yang, MC
Hwang, MT
Tsao, JH
Mao, SJT
生物科技學系
Department of Biological Science and Technology
關鍵字: beta-lactoglobulin structure;immunoassay native monoclonal antibody;thermal denaturation
公開日期: 1-三月-2006
摘要: Molten globules are thought to be general intermediates in protein folding and unfolding. beta-lactoglobulin (beta-LG) is one of the major bovine whey proteins, constituting similar to 10 to 15% of total milk proteins. We have recently identified beta-LG as a superior marker for evaluating thermally processed milk. Strand D of beta-LG participates in irreversible thermal unfolding as probed by a monoclonal antibody (mAb) specific to thermally denatured beta-LG. In the present study, we used native beta-LG as an immunogen to test the hypothesis that a specific mAb against the native beta-LG could be established. As result, a mAb (4H11E8) directed against the native structure of beta-LG was made. The antibody did not recognize the heat-denatured form of beta-LG, such as its dimer and aggregates. Immunoassay using this "native" mAb showed that the stability of beta-LG was at temperatures = 70 degrees C. beta-Lactoglobulin began to deteriorate between 70 and 80 degrees C over time. The denaturation was correlated with the transition temperature of beta-LG. Further chemical modification of Cys (carboxymethylation) or positively charged residues (acetylation) of beta-LG totally abolished its immunoreactivity, confirming the conformation-dependent nature of this mAb. Using competitive ELISA, the 4H11E8 mAb could determine the native beta-LG content in commercially processed milks. Concentrations of native beta-LG varied significantly among the local brands tested. From a technological standpoint, the mAb prepared in this study is relevant to the design and operation of appropriate processes for thermal sanitation of milk and of other dairy products.
URI: http://hdl.handle.net/11536/12513
ISSN: 0022-0302
期刊: JOURNAL OF DAIRY SCIENCE
Volume: 89
Issue: 3
起始頁: 912
結束頁: 921
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