三种蛋白含量测定试剂盒检测微量蛋白的性能比较
Comparison of performance of three protein content determination kits for detecting micro protein
曹明翔 1徐华 1刘碧云 1张浩然 1姜莉 1杨二霞1
作者信息
- 1. 艾美行动生物制药有限公司,泰州 225300
- 折叠
摘要
目的 比较三种蛋白含量测定试剂盒(艾美Lowry法、康为BCA法、赛默飞BCA法)检测微量(0～20 µg/mL)蛋白的性能差异,选出最准确可靠的试剂盒用于微量蛋白样品检测.方法 分别使用该三种试剂盒检测微量蛋白含量,比较其准确度、灵敏度、线性及重复性,并评价其等效性.结果 三种试剂盒检测微量(1.25～20 µg/mL)蛋白的回收率均在75%～120%范围内,表现出合格的准确度;赛默飞BCA在2.5～20 µg/mL,艾美Lowry和康为BCA在5～20 µg/mL范围内符合精密度可接受范围(RSD≤6%),表现出较好的重复性;三种试剂盒标准曲线(0～20 µg/mL)的相关系数(R2)均高于 0.980,艾美Lowry线性的略差为 0.981,康为BCA的线性合格为 0.991,赛默飞BCA的线性最佳为0.993;以上方法学验证结果得出三种试剂盒的灵敏度的最低定量限分别为艾美Lowry法5 µg/mL、康为BCA法 5 µg/mL、赛默飞BCA法 2.5 µg/mL.三种试剂盒对同一系列浓度BSA检测的值两两配对作曲线,其相关性系数均高于 0.990.结论 通过实验,我们发现各试剂盒在不同浓度BSA样品中表现出不同的性能,其中赛默飞BCA法在准确度和重复性方面表现最优,且三种方法之间存在一定的相关性.
Abstract
Objective Compare the performance differences of three protein content determination kits(Lowry method of Aimei,BCA method of Kangwei,and BCA method of Thermo Fisher)in detecting micro(0～20 µg/mL)protein.Select the most accurate and reliable reagent kit for the detection of trace protein samples.Methods Use these three test kits to detect micro protein content,compare their accuracy,sensitivity,linearity and repeatability,and evaluate their equivalence.Results The recovery rates of the three test kits for detecting trace amounts(1.25～20 µg/mL)of protein were all within the range of 75%～120%,demonstrating qualified accuracy;Thermo Fisher BCA meets the acceptable precision range(RSD≤6% )in the range of 2.5 µg/mL to 20 µg/mL,while Aimei Lowry and Kangwei BCA meet the acceptable precision range(RSD≤6% )in the range of 5 µg/mL to 20 µg/mL,demonstrating good repeatability;the correlation coefficient R2 of the standard curves of the three test kits(0～20 µg/mL)are all higher than 0.980.The linearity of Aimei Lowry is slightly poor,with R2 of 0.981.The linearity of Kangwei BCA is qualified,with R2 of 0.991.The linearity of Thermo Fisher BCA is the best,with R2 of 0.993;The above methodological validation results indicate that the minimum quantification limits for the sensitivity of the three test kits are 5 µg/mL for the Aimei Lowry method,5 µg/mL for the Kangwei BCA method,and 2.5 µg/mL for the Thermo Fisher BCA method,respectively;Paired curves of values detected by three reagent kits for the same series of BSA concentrations and their correlation coefficients were all higher than 0.990.Conclusion Through experiments,we found that each reagent kit exhibited different performance in different concentrations of BSA samples,among which the Thermo Fisher BCA method performed the best in terms of accuracy and repeatability,and there was a certain correlation between these three methods.
关键词
蛋白含量测定试剂盒/准确度/灵敏度/重复性/等效性Key words
protein content determination kit/accuracy/sensitivity/repeatability/equivalence引用本文复制引用
出版年
2024
NETL
NSTL
万方数据