Irurzun P L, Miranda M P
Allergol Immunopathol (Madr). 1976 Mar-Apr;4(2):111-20.
Several components of catodic electrophoretic migration in serum and urine are present in normal individuals and in rabbit serum. There also exists in man and in some animals, serum fractions of low molecular weight. These types of serum components may be or may not be related with the IgG. In a previous study we have isolated two components in the slow catodic electrophoretic area of the normal human serum (NHS). One of them was identified as an IgG subclass and the other component presented a clear line of precipitation to gamma heavy chain specific immuno-serum. This latter component was found in the post gamma-globulin area crossing the IgG arc in the I.E. analysis. Its molecular weight was variable from 3700 to 9500. In this paper a differential analysis of the gamma fragment isolated for us, is made and its relationship with Fc subfragments of pepsin-digested IgG is studied. In order to obtain this comparative study, the electrophoresis, gel diffusion immunoelectrophoresis gel chromatography and analytic ultracentrifugation techniques are employed. The post-gammaglobulin fraction has been isolated from total normal human serum without previous manipulation, or with the gammaglobulin fraction precipitated with saturated ammonium sulphate, in Sephadex G-200 chromatography. These two fractions present similar immunelectrophoretical characteristics. The constant sedimentation is 0.90 S and the approximated molecular weight is 7000. Since the pepsin digestion of IgG produced Fc subfragments of low molecular weight, we have isolated and submitted this immunoglobulin to peptic digestion. The G-75 Sephadex filtration shows an isolated post-gamma-globulin of I.E. sedimentation constant and molecular weight whose characteristics are similar to the isolated serum post-gammaglobulin fraction. The antigenical analysis in I.D. shows a total identity between the pepsin digested post-gammaglobulin and the fragment obtained for us from the human serum to an anti-heavy gamma chain immuno-serum and to a rabbit serum anti-fraction produced for us. These events suggest that this isolation post-gamma-globulin fraction corresponds to a Fc subfragment of the IgG.
正常人和兔血清中存在血清及尿液中阴极电泳迁移的几种成分。人和一些动物体内还存在低分子量的血清组分。这些血清成分类型可能与IgG有关,也可能无关。在之前的一项研究中,我们从正常人血清(NHS)的慢速阴极电泳区域分离出两种成分。其中一种被鉴定为IgG亚类,另一种成分与γ重链特异性免疫血清呈现出清晰的沉淀线。在免疫电泳分析中,后一种成分出现在γ球蛋白区之后,穿过IgG弧线。其分子量在3700至9500之间变化。本文对为我们分离出的γ片段进行了差异分析,并研究了其与胃蛋白酶消化的IgG的Fc亚片段的关系。为了进行这项比较研究,采用了电泳、凝胶扩散免疫电泳、凝胶色谱和分析超速离心技术。后γ球蛋白组分是从未经预先处理的正常人血清中分离出来的,或者是在Sephadex G - 200色谱中用饱和硫酸铵沉淀γ球蛋白组分后分离得到的。这两种组分呈现出相似的免疫电泳特征。恒定沉降系数为0.90 S,近似分子量为7000。由于IgG经胃蛋白酶消化会产生低分子量的Fc亚片段,我们分离了这种免疫球蛋白并对其进行胃蛋白酶消化。G - 75 Sephadex过滤显示出一种分离的具有免疫电泳沉降系数和分子量的后γ球蛋白,其特征与分离的血清后γ球蛋白组分相似。免疫扩散中的抗原分析表明,胃蛋白酶消化的后γ球蛋白与我们从人血清中获得的片段,对于抗γ重链免疫血清和为我们制备的兔抗组分血清来说,具有完全一致性。这些结果表明,这种分离的后γ球蛋白组分对应于IgG的一个Fc亚片段。