溶血素类型和含量的分光光度测定法

徐学瑛等建立的溶血素测定法已在国内免疫药理学研究中得到广泛应用。溶血素主要由IgG和IgM组成,由于IgG溶血效率大大低于IgMC单独结合绵羊红细胞(Sheep redblood cell,SRBC)不足以活化补体产生溶血,所以原法只能测定IgM溶血素。根据2-巯基乙醇(2-mercaptoethanol,2-ME)能够破坏IgM抗体而保留IgG抗体活性的性质,利用抗IgG血清使IgG交联后产生溶血的原理,本文报告一个能同时测定IgG、IgM溶血素和总溶血素含量的分光光度法,并用几种已知药物对该方法进行了检验。

SPECTROPHOTOMETRIC DETERMINATION OF HEMOLYSIN TYPES AND CONCENTRATION

A quanitiative method for measuring IgM, IgG and total hemolysin in serum is described. Immune mice serum (nontreated for determing IgM or treated with equal volume of 2-mercaptoethanol 0.2 mol/L for IgG). Sheep red blood cells (SRBC), guinea pig complement and-where applicable-sheep antimice immunoglobulin G serum were incubated at 37℃ for lh in fluid phase, centrifuged, and the supernatant read at 540 nm. Hemolysin concentration was calculated by OD₅₄₀×dilution of serum.The IgG, IgM and total hemolysin concentrations in mice treated with hydrocortisone, cyclophosphamide or antilymphocyte serum (ALS) were decreased, but those of Bacille Calmette-Guerin (BCG) or levamisole treated mice were increased.It seems that this method is simple, rapid and exact.