摘要
目的:建立氢氧化铝吸附重组肺炎蛋白疫苗原液及制剂的解吸附方法。方法:在各重组肺炎蛋白的氢氧化铝吸附原液或4组分疫苗制剂半成品中加入解吸附试剂[含0.4%(m/v)十二烷基二甲基甜菜碱(BS-12),0.25 mo L·L-1的磷酸盐缓冲液,p H 6.5],37℃下静置4.5 h,通过Lowry蛋白检测、SDS-PAGE和HPLC对蛋白的解吸附率进行测定。结果:结果显示该方法可将重组肺炎蛋白疫苗的4种蛋白从氢氧化铝佐剂上解吸附下来,解吸附率达95%以上,且不影响蛋白的完整结构。结论:建立了重组肺炎蛋白疫苗4个组分蛋白的氢氧化铝吸附原液及制剂半成品的解吸附方法,可用于疫苗吸附原液和制剂半成品定量分析的前处理。
Objective： To develop a method for the desorption of four recombinant pneumococcal proteins from the aluminum hydroxide （AH） adjuvant in a tetravalent protein based pneumoeoccal vaccine （PBPV） prepa- ration. Methods： The PBPV stock solution or final vaccine formulation samples were treated in the desorption solu- tion, which contains 0.4% amphiprotic surfactant （ BS-12 ） in 0.25 moL· L- 1 phosphate buffer （ pH 6.5 ） for 4.5 h at 37 ℃. The proteins desorbed from the AH adjuvant were then analyzed using Lowry protein assay, SDS-PAGE and HPI,C. Results： A desorption rate of 95% or higher was achieved, and the procedure was shown to have no impact on the integrity of the PBPV antigens. Conclusion： The PBPV antigens in the vaccine stock solution and vaccine formulation can be easily and efficiently desorbed from the AH adjuvant using the method developed in this study. The procedure is suitable for the pretreatment of the PBPV vaccine preparations before antigen content deter- minatiun.
出处
《中国新药杂志》
CAS
CSCD
北大核心
2015年第9期998-1001,1023,共5页
Chinese Journal of New Drugs
基金
国家"重大新药创制"科技重大专项(2014ZX09102046)
