Patent ID: 9645156
Date: 2017-05-09
CPC Classifications: G01N,Y10T

Claim:
1. A method for determining sample glycosylation and terminal modifications during purification process of immunoglobulin, consisting of the following steps of: Step 1) separating immunoglobulin by using cation-exchange chromatography, and collecting different components in accordance to retention time; Step 2) denaturing the components of immunoglobulin obtained in step 1) with a denaturant, followed by reducing the components with a reducing agent, to separate light chain and heavy chain of the immunoglobulin; Step 3) separating the light chain and the heavy chain of immunoglobulin of step 2) by using C4 reverse phase ultrahigh pressure liquid chromatography having a mobile phase X of 0.1% formic acid in water and mobile phase Y of 0.1% formic acid in acetonitrile to achieve a baseline separation of the light and the heavy chains; Step 4) measuring molecular weights of the light chain and the heavy chain obtained in step 3) with a mass spectrometer; and Step 5) analyzing chromatographic data obtained in step 3) and mass spectrometric data obtained in step 4) to determine glycosylation and terminal modifications of said immunoglobulin.