蛋白A高效亲和膜色谱法测定人血浆中免疫球蛋白G的含量

 研究了蛋白Ａ高效亲和膜色谱对水溶液及人血浆中人免疫球蛋白Ｇ（ＨＩｇＧ）的特异性吸附和定量测定。方法具有较高的精确度和较好的重复性：ＨＩｇＧ标样５次重复进样的相对标准偏差为１．５％，人血浆样品３次重复进样的相对标准偏差为３．６％；所测得的定量标定曲线的线性相关系数达到０．９９９３；不含己二胺间隔臂的亲和介质的非特异性吸附极低，基本检测不出来。快速实验中，一次分析可在０．５ｍｉｎ内完成。实验表明，利用所建立的方法对人血浆中的ＨＩｇＧ进行定量测定可以得到较为满意的结果。

Determination of HIgG in Human plasma by high performance membrane affinity chromatography(HPMAC)]

An assay technique for determination of Human IgG in human plasma has been developed by utilizing Protein A bound to the modified cellulose matrices based on the strong affinity between protein A and Fc region of IgG. The pH values of loading buffer and elution buffer were 7.0 and 2.3 respectively. The cartridge used was 20 mm x 4 mm i.d. and detection was carried out with a UV monitor at 280 nm. The non-specific adsorption of two kinds of media has been studied. The media without hexyldiamine arm gives very low non-specific adsorption BSA. The calibration curve showed a good linearity (correlation coefficient > 0.9992) in the injected amount range of 9-70 micrograms for HIgG. The total time for separation and determination was within 5 min and the total time of rapid assay was within 30 s. Relative standard deviations of peak areas were 1.5% (n = 5) and 3.6% (n = 3) for HIgG in standard solution and human plasma respectively.