A Novel Method for in Situ Detection of Hydrolyzable Casein Fragments in a Cheese Matrix by Antibody Phage Display Technique and CLSM
作者： Zhi Duan , Dagmar Adeline Brüggemann , Henrik Siegumfeldt
DOI: 10.1021/JF901026M
关键词: Casein 、 Biology 、 Biochemistry 、 Confocal microscopy 、 Western blot 、 Bacteriophage 、 Cheese ripening 、 Peptide 、 Phage display 、 In situ
摘要: A novel method to monitor in situ hydrolyzable casein fragments during cheese ripening by using immunofluorescent labeling and confocal laser scanning microscopy (CLSM) was developed. Monoclonal single chain variable of antibody (scFvs) were generated phage display toward three small synthetic peptides the α s1 -casein sequence. These traverse enzymatic cleavage sites ripening. The specificity antipeptide antibodies determined ELISA Western blot. Finally, an protocol successfully developed for detection scFvs binding different inside a matrix CLSM. To our knowledge, this is first demonstrated analysis proteolysis phenomena matrix. Additionally, technique offers high potential study dynamic spatial changes target components complex food systems.
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