Johnson R B, Libby R
J Clin Microbiol. 1980 Sep;12(3):451-4. doi: 10.1128/jcm.12.3.451-454.1980.
The proposed method was designed to replace the tedious and difficult separation of immunoglobulin M (IgM) from IgG by sucrose gradient sedimentation. In this method, a 250-microliter portion of serum diluted 20-fold was passed through a small column of quaternary aminoethyl-Sephadex A-50 ion exchanger. IgG was not retained, but additional washes were required to remove all but 5%. A second buffer-eluting fluid recovered an average of 80% of the original IgM in a defined dilution. The entire operation took 15 min. The efficiency of this process was evaluated by the following: (i) radial immunodiffusion measurements of IgG and IgM; (ii) recovery studies of isohemagglutinins; and (iii) demonstrated removal of interference by the rheumatoid factor. The method was applied successfully to distinguish rubella IgM antibody.
所提出的方法旨在取代通过蔗糖梯度沉降从IgG中繁琐且困难地分离免疫球蛋白M（IgM）的过程。在该方法中，将250微升稀释20倍的血清通过一小柱季胺乙基 - 葡聚糖凝胶A - 50离子交换剂。IgG未被保留，但需要额外洗涤以除去除5％以外的所有物质。第二种缓冲洗脱液以规定的稀释度平均回收了原始IgM的80％。整个操作耗时15分钟。该过程的效率通过以下方式评估：（i）对IgG和IgM进行径向免疫扩散测量；（ii）对同种血凝素的回收率研究；以及（iii）证明类风湿因子的干扰已被消除。该方法成功应用于区分风疹IgM抗体。
