Harrison H H, Miller K L, Dickinson C, Daufeldt J A
Department of Pathology, University of Chicago Pritzker School of Medicine, Illinois 60637.
Am J Clin Pathol. 1992 Jan;97(1):97-105. doi: 10.1093/ajcp/97.1.97.
Qualitative data from high-resolution two-dimensional electrophoresis (2DE) have become clinically useful in immunoglobulin clonality analysis, in resolution of ambiguities in immunofixation typing of paraproteins, and in genetic typing of serum proteins. Since 1986, the authors have been evaluating the College of American Pathologists Reference Preparation for Serum Proteins (RPSP) as a quality control material for 2DE because (1) it is prepared exclusively from pooled human sera, (2) the pool yields a reference pattern of mixed heterozygosity for genetic markers, and (3) RPSP is widely available as a lyophilized preparation that currently serves in the authors' laboratory as a qualitative quality control preparation and that may become a quantitative quality control material or external quality assessment material for 2DE. Using the ISO-DALT 2DE system and silver-staining, the peptide patterns were examined in 11 lots of RPSP and compared with fresh serum and with each other. Consistent differences in the 2DE pattern between RPSP and fresh serum included the presence of freeze-thaw peptides, the presence of degradation spots of apolipoprotein A-I, and the diminution of apolipoprotein spot intensities in RPSP. All lots of RPSP yielded clear identification of the eight serum proteins used for quality control calculations. Run-to-run coefficients of variation for a single lot of RPSP for four parameters of 2DE spot location and gradient reproducibility were comparable with band location reproducibility for the one-dimensional procedures of serum protein electrophoresis and lactate dehydrogenase isoenzyme electrophoresis. It is concluded that the reproducibility, that is, imprecision, of 2DE is the same as one-dimensional clinical electrophoresis techniques and that either RPSP or pooled fresh serum can serve as a satisfactory internal quality control material.
来自高分辨率二维电泳（2DE）的定性数据在免疫球蛋白克隆性分析、副蛋白免疫固定分型中的模糊性解析以及血清蛋白基因分型方面已具有临床实用性。自1986年以来，作者一直在评估美国病理学家学会血清蛋白参考制剂（RPSP）作为2DE的质量控制材料，原因如下：（1）它完全由混合人血清制备；（2）该混合血清产生了遗传标记的混合杂合性参考图谱；（3）RPSP以冻干制剂形式广泛可得，目前在作者实验室用作定性质量控制制剂，并且可能成为2DE的定量质量控制材料或外部质量评估材料。使用ISO-DALT 2DE系统和银染法，对11批RPSP的肽图谱进行了检测，并与新鲜血清以及彼此进行比较。RPSP和新鲜血清在2DE图谱上的一致差异包括冻融肽的存在、载脂蛋白A-I降解斑点的存在以及RPSP中载脂蛋白斑点强度的降低。所有批次的RPSP都能清晰鉴定用于质量控制计算的8种血清蛋白。对于RPSP的单个批次，2DE斑点位置和梯度重现性的四个参数的批间变异系数与血清蛋白电泳和乳酸脱氢酶同工酶电泳的一维程序的条带位置重现性相当。结论是，2DE的重现性，即不精密度，与一维临床电泳技术相同，并且RPSP或混合新鲜血清均可作为令人满意的内部质量控制材料。
