A Novel Method of the Enzyme Immobilization for the Biological Applications

摘要

Enzyme immobilization refers to the process of attaching free or soluble enzymes to an inert or insoluble support, which allows the enzyme to be re-used and physically separates the product with great ease. In this study, a straightforward and reliable enzyme immobilization method is developed based on the polymerization of acrylamide together with the enzyme of interest. This approach is well coupled with relative and absolute quantification by mass spectrometry, as shown by the example of enzymatic ~(16)O/~(18)O labeling and the preparation of AQUA-like peptides, respectively. Furthermore, the ~(16)O/~(18)O labeling disulfide bridge peptides can be selectively detected by means of mass pair detection in LTQ-Orbitrap mass spectrometer by using this strategy.