PROPERTIES OF UNIVALENT FRAGMENTS OF RABBIT ANTIBODY ISOLATED BY SPECIFIC ADSORPTION
PROPERTIES OF UNIVALENT FRAGMENTS OF RABBIT ANTIBODY ISOLATED BY SPECIFIC ADSORPTION
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DOI：
10.1016/0003-9861(60)90229-0
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发表时间：
1960-01-01
影响因子：
3.9
通讯作者：
WOERNLEY, DL
中科院分区：
文献类型：
--
作者：
NISONOFF, A;WISSLER, FC;WOERNLEY, DL
描述了一种通过特异性吸附分离兔抗半抗原抗体活性非沉淀片段的方法。这些片段大约是原始分子的 1/3，是从专门纯化的抗对偶氮苯甲酸酯抗体的木瓜蛋白酶水解产物中分离出来的。它们与同源半抗原相互作用的平均平衡常数和异质性指数在实验误差范围内与未处理抗体的值相同。因此，分离程序不会根据分子的结合亲和力来分级分子。同样明显的是，2/3 的分子可以被丢弃，而不影响结合位点的结构完整性，并且由于分子的剩余体积而对结合的静电效应可以忽略不计。通过半抗原结合测量确定，分离片段的每个分子的平均结合位点数量为 0.9-1.0，这提供了木瓜蛋白酶产生的活性片段是单价的直接证据。透析的木瓜蛋白酶水解物的比旋光度与未处理的兔γ-球蛋白的比旋光度大致相同，表明片段是紧凑的分子单元； i.e., are not appreciably unfolded or denatured.伴随水解的比粘度的降低和热稳定性不变也支持了这一结论。
A method is described for the isolation of active nonprecipitating fragments of rabbit antihapten antibody by specific adsorption. The fragments, about 1/3 as large as the original molecule, were isolated fror a papain hydrolysate of specifically purified anti-p-azobenzoate antibody. The average equilibrium constant for their interaction with homologous hapten, and the index of heterogeneity were the same, within experimental error, as the values for untreated antibody. Thus the isolation procedure does not fractionate molecules according to their combining affinity. It is also evident that 2/3 of the molecule may be discarded without affecting the structural integrity of the combining site and that electrostatic effects on binding due to the residual bulk of the molecule are negligible. The average number of combining sites per molecule of the isolated fragments, determined by hapten-binding measurements, is 0.9-1.0, providing direct evidence that the active fragments produced by papain are univalent. The specific optical rotation of a dialyzed papain hydrolyzate is approximately the same as that of untreated rabbit [gamma] -globulin, indicating that the fragments are compact molecular units; i.e., are not appreciably unfolded or denatured. This conclusion is supported by the reduction in specific viscosity and the unchanged heat stability which accompanies the hydrolysis.