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(0~20μg/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(1.25~20μg/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2.5~20μg/mL,艾美Lowry和康为BCA在5~20μg/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(0~20μg/mL)的相关系数(R^(2))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5μg/mL、康为BCA法5μg/mL、赛默飞BCA法2.5μg/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。
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(0~20μg/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(1.25~20μg/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 2.5μg/mL to 20μg/mL,while Aimei Lowry and Kangwei BCA meet the acceptable precision range(RSD≤6%)in the range of 5μg/mL to 20μg/mL,demonstrating good repeatability;the correlation coefficient R^(2)of the standard curves of the three test kits(0~20μg/mL)are all higher than 0.980.The linearity of Aimei Lowry is slightly poor,with R^(2)of 0.981.The linearity of Kangwei BCA is qualified,with R^(2)of 0.991.The linearity of Thermo Fisher BCA is the best,with R^(2)of 0.993;The above methodological validation results indicate that the minimum quantification limits for the sensitivity of the three test kits are 5μg/mL for the Aimei Lowry method,5μg/mL for the Kangwei BCA method,and 2.5μg/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.
作者
曹明翔
徐华
刘碧云
张浩然
姜莉
杨二霞
CAO Ming-Xiang;XU Hua;LIU Bi-Yun;ZHANG Hao-Ran;JIANG Li;YANG Er-Xia(Aimei Action Biopharmaceutical Co.,Ltd.,Taizhou 225300,China)
出处
《实验室检测》
2024年第7期126-130,共5页
Laboratory Testing
关键词
蛋白含量测定试剂盒
准确度
灵敏度
重复性
等效性
protein content determination kit
accuracy
sensitivity
repeatability
equivalence
