Stemberger H, Scheiner O, Wiedermann G, Kraft D, Förster O
J Immunol Methods. 1978;21(3-4):365-72. doi: 10.1016/0022-1759(78)90163-1.
Affinity chromatography and subsequent ion-exchange chromatography of pooled anti-benzylpenicilloyl (anti-BPO) hyperimmune sera separated 5 different anti-BPO IgG fractions as described in the preceding paper. These fractions were tested for activities in passive hemagglutination (PHA), passive immune hemolysis (PIH), antibody-dependent cellular cytotoxicity (ADCC) and influence on IgM-induced hemolysis. It was found that anti-BPO IgG fractions with low avidity (dissociation constant K = 4.4--6.7 X 10(-8) moles/l) were poorly active in PHA and ADCC and had no blocking activity in IgM hemolysis. Among the highly avid antibodies (K = 0.7--3.4 X 10(-8) mole/l) no correlations were found between avidity and activities in the immunological tests. The results presented demonstrate that ion-exchange chromatography allows the separation of blocking and lytic antibodies as shown by their influence on IgM-induced complement-dependent lysis of lightly hapten-coated sheep erythrocytes.
如前文所述，对汇集的抗苄青霉素酰基（抗BPO）超免疫血清进行亲和层析及随后的离子交换层析，分离出了5种不同的抗BPO IgG组分。对这些组分进行了被动血凝（PHA）、被动免疫溶血（PIH）、抗体依赖性细胞毒性（ADCC）活性以及对IgM诱导溶血的影响的检测。结果发现，亲和力低（解离常数K = 4.4--6.7×10⁻⁸摩尔/升）的抗BPO IgG组分在PHA和ADCC中活性较差，且对IgM溶血无阻断活性。在高亲和力抗体（K = 0.7--3.4×10⁻⁸摩尔/升）中，未发现亲和力与免疫检测中的活性之间存在相关性。所呈现的结果表明，离子交换层析能够分离阻断性抗体和溶细胞性抗体，这可通过它们对IgM诱导的轻度半抗原包被绵羊红细胞补体依赖性溶解的影响来体现。
