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 |
顯示於類別: | 期刊論文 |