Howerton D A, Check I J, Hunter R L
Am J Clin Pathol. 1986 Feb;85(2):213-8. doi: 10.1093/ajcp/85.2.213.
The authors developed a cost-effective procedure for high-resolution protein electrophoresis in agarose gel which results in electrophoretic patterns that can be both visually interpreted and densitometrically quantitated. Unique features include the development of special molds for the preparation of several gels simultaneously and the combination of chemical with densitometric procedures to accurately quantitate the protein fractions in both normal and pathologic sera. Since the albumin concentrations obtained by densitometric scan of either cellulose acetate or agarose gel were erratic with respect to colorimetric (ACA) or immunochemical (ICS) measurements, the authors incorporated colorimetric measurements of albumin and total protein with densitometric quantitation of the electrophoretic bands into an algorithm for calculating protein concentrations. The densitometric quantitation of the gamma globulin fractions compared well with the sum of the immunochemically determined IgG and IgM concentrations (r = 0.98). Alpha and beta globulins were calculated using mathematically adjusted densitometric scan percentages and the total alpha and beta globulin protein concentration. The agarose gel procedure was more sensitive than cellulose acetate in the detection of paraprotein bands in a comparison of 93 sera containing paraproteins.
作者开发了一种用于琼脂糖凝胶中高分辨率蛋白质电泳的经济高效方法，该方法产生的电泳图谱既可以直观解读，也可以进行光密度定量分析。其独特之处包括开发了用于同时制备多种凝胶的特殊模具，以及将化学方法与光密度分析程序相结合，以准确测定正常和病理血清中的蛋白质组分。由于通过醋酸纤维素或琼脂糖凝胶的光密度扫描获得的白蛋白浓度与比色法（ACA）或免疫化学法（ICS）测量结果不一致，作者将白蛋白和总蛋白的比色测量与电泳条带的光密度定量纳入一个计算蛋白质浓度的算法中。γ球蛋白组分的光密度定量与免疫化学测定的IgG和IgM浓度之和相比效果良好（r = 0.98）。使用数学调整后的光密度扫描百分比和总α和β球蛋白蛋白质浓度来计算α和β球蛋白。在对93份含副蛋白的血清进行比较时，琼脂糖凝胶方法在检测副蛋白条带方面比醋酸纤维素更敏感。
