摘要
目的:优化现有的甲胎蛋白异质体(AFP-L3)分离和检测方法,找到一个最有效的AFP-L3的提取方法,为今后的AFP-L3研究奠定前期基础。方法:应用北京热景生物技术公司的甲胎蛋白异质体(AFP-L3)亲和吸附离心管,通过改变亲和介质使用量和洗脱液体积的配伍梯度变化来优化实验流程,排除提取AFP-L3的影响因素,分离血清中的AFP-L3,并用罗氏公司的甲胎蛋白检测试剂盒检测分离的效果。结果:发现在不改变亲和介质体积的情况下,随着洗脱液体积的减少AFP-L3回收浓度逐渐增加,且在75 ul洗脱液时能够获得最高AFP-L3回收浓度。结论:成功地优化了北京热景生物技术公司的甲胎蛋白异质体(AFP-L3)亲和吸附离心管及其配套试剂的使用方法。
Objective： To optimize the existing method of the heterogeneity of AFP separation and develop the most effective AFP-L3 extraction method for further research. Methods： The experimental process of Heterogeneity of AFP （AFP-L3） affinity adsorption tubes produced by Beijing Hotgen biotechnology company was optimized by changing the affinity media compatibility and usage gradient elution volume. AFP-L3 was separated in the serum and expelling other factors which can interfere the separation. The results have been favourably proven by Roche fetoprotein test kit. Results： Keeping the affinity media compatibility volume unchanged, the concentration of AFP-L3 increased with the decreasing of washing buffer volume. The concentration of AFP-L3 was the highest when the washing buffer volume was 75 ul. Conclusion： Successfully optimized the method of the heterogeneity of alpha-fetoprotein （AFP-L3） adsorption affinity reagent tubes produced Beijing Hotgen biotechnology company.
出处
《现代生物医学进展》
CAS
2014年第24期4630-4633,共4页
Progress in Modern Biomedicine
基金
北京市肝病研究所2012年度双十计划项目(Z121107002812113)
北京市肝病研究所基金(BJIH-01206)
首都医科大学重点实验室基金(2013GYGA03)
国家国际科技合作专项<中美英联合乙肝相关肝癌早期诊断分子标志物筛查及免疫机制研究>(S2012ZR0153)
