HBsAg生化性状对乙型肝炎疫苗效力影响的研究

Impacts of HBsAg biochemical forms on the potencies of the recombinant yeast hepatitis B vaccines

比较研究了重组酵母表达HBsAg的P2 4亚基间二硫键结合及其还原条件下降解程度对乙型肝炎疫苗效力的影响。分别用HPLC及SDS PAGE法检测重组酵母乙型肝炎疫苗HBsAg的P2 4亚基在还原条件下降解和P2 4亚基间二硫键桥联程度 ,并检测乙型肝炎疫苗小鼠体内效力 (ED50 值 )和体外相对效力 (RP) ,分析HBsAg的P2 4亚基间二硫键结合及其还原条件下降解程度对乙型肝炎疫苗效力的影响。结果表明HBsAgP2 4亚基还原条件下降解成分Sub P2 4的相对含量在 5 .2 %～ 31%间、P2 4亚基间二硫键桥联型HBsAg颗粒相对含量在 41.9%～ 71.8%范围内与疫苗的实验室效力无关。所生产重组酵母乙型肝炎疫苗HBsAg的P2

For comparing and studying the impacts of the reductive extent of P24 peptide (Sub-P24)of HBsAg and the disulfide-bonded degree of P24 unit of HBsAg on the laboratory potencies of the recombinant yeast hepatitis B vaccines, the percentages of the reduced P24 peptide (Sub-P24) derived from the P24 with the treatment of DTT and boiling and the particle form HBsAg composed of its disulfide-bonded P24 polypeptide unit were detected by silver stained SDS-PAGE and HPLC respectively. The in vivo mouse potency (ED 50) and in vitro relative potency(RP)for 109 lots of the recombinant yeast hepatitis B vaccines were also tested. The influence of proportion of disulfide-bonded particle HBsAg and Sub-P24 on the potency of the vaccines was analyzed. The results show that the vaccine potency is not impacted by the variations of ratio for the Sub-P24 within 5.2%～31% range and the disulfide-bonded HBsAg from 41.9% to 71.8%. The maximum lot-differences of ED 50 and RP are 10.3 and 2.2 folds respectively. It suggests that the disulfide-bonded degree of P24 unit of HBsAg and its reduction of P24 peptide(Sub-P24) did not affect the potencies of the HB vaccines.