Method for the determination of the proportion of a fusion protein in a mixture of fusion protein and proteolytic degradation products

摘要

In the method, the mixture of fusion protein and degradation product is diluted with bovine serum albumin. One portion of the diluted mixture is mixed with alpha -amylase solution, incubated at 37 DEG C for /= 30 minutes, alpha -amylase reagent is added and the remaining activity in the alpha -amylase solution is measured by spectrophotometry. The other portion of the diluted mixture is mixed with alpha -amylase reagent and, after mixing, alpha -amylase solution is added and the enzyme kinetics (course of the reaction) are measured by spectrophotometry. All the measurements are compared with standards of fusion protein, tendamistat, degradation product and uninhibited alpha -amylase reaction, and the content of fusion protein in the mixture is determined from the comparison. IMAGE