重组溶葡萄球菌酶中宿主菌残余蛋白含量的测定

Determination of Residual B.Sphaericus Protein in Recombinant Lysostaphin Products

制备球形芽孢杆菌菌体蛋白作为免疫抗原,免疫家兔制备抗血清,建立了一种测定重组溶葡萄球菌酶中残余宿主菌菌体蛋白含量的双抗体夹心酶联免疫吸附测定法(Enzyme-linked immunosorbent assay,ELISA)。结果表明,建立的双抗体夹心ELISA法快速、简便、灵敏,检测范围宽,能检测出制品中的微量菌体蛋白;对3批重组溶葡萄球菌酶原液的检测合格,制品中残余宿主菌蛋白含量小于万分之二。此方法可以用于重组溶葡萄球菌酶中残余宿主菌菌体蛋白含量的测定。

To establish a double antibody sandwich ELISA method for determination of residual B. sphaericus protein in recombinant lysostaphin products, the antiserum against the B. sphaericus protein was prepared by immunizing rabbits. The sensitivity of the method was about 1.0 ng/ml. A good linearity was observed for B. sphaericus protein in the range of 7. 8-125 ng/ml. The average recovery of B. sphaericus protein was 98.77% with RSD 11.08%. Based on this method, 3 batches of recombinant lysostaphin products were tested, and the content of B. sphaericus protein in the sample was less than 0.02%. The method is simple, sensitive and accurate, so the trace of bacteria protein in recombinant lysostaphin products can be detected.