张爱英 陈怡 侯传浩 汪志鹏 乔录新 杨国申 杨世申 李宁 陈德喜.甲胎蛋白异质体分离方法的优化[J].,2014,14(24):4630-4633 |
甲胎蛋白异质体分离方法的优化 |
The Optimization of Alpha-fetoprotein Variants(AFP-L3) Test |
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DOI: |
中文关键词: 亲和吸附 甲胎蛋白异质体 甲胎蛋白检测 |
英文关键词: Affinity adsorption Fetoprotein heterogeneity AFP test |
基金项目: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) |
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中文摘要: |
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。 |
英文摘要: |
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. |
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