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尝试通过亲和层析法分离抗红细胞抗体。二、IgG——不完全抗体。

Attempt to isolate antierythrocyte antibodies by the method of affinity chromatography. II. IgG - incomplete antibodies.

作者信息

Simková J

出版信息

J Hyg Epidemiol Microbiol Immunol. 1979;23(1):104-9.

PMID:574883
Abstract

It ensues from the results of the present work that erythrocyte stromata can serve as a suitable insoluble immunosorbent of both complete and incomplete antibodies. When applying optimum conditions, such as laboratory incubation temperature and suitable buffers, it is possible, by means of a chromatographic column with 0.5 ml erythrocyte stromata suspended in 40 g of glass beads (ballotini), to obtain fractions devoid of incomplete antibodies. Incomplete antibodies could be absorbed completely, either directly by a 24-hour incubation of the respective serum on the column or by passaging the serum through the column. In the presence of 0.2 M NaCl, absorption of incomplete antibodies was prevented; only complete antibodies were demonstrated in the fractions of the eluates. The elution yield in incomplete antibodies was low; in some cases, it could not be demonstrated at all. It can be explained partly by non-specific absorption, partly by the low initial antibody titre. It is therefore rather difficult to separate the individual incomplete antibodies from each other in this method. The column with incorporated erythrocyte stromata can be recommended for incomplete antibodies only because of its absorption capacity (to remove incomplete antibodies), but not for obtained elution yield.

摘要

从目前的工作结果可以看出,红细胞基质可作为完整抗体和不完全抗体的合适不溶性免疫吸附剂。在应用最佳条件时,如实验室孵育温度和合适的缓冲液,通过将0.5 ml红细胞基质悬浮于40 g玻璃珠(玻璃粉)中的色谱柱,有可能获得不含不完全抗体的组分。不完全抗体可以通过将相应血清在柱上直接孵育24小时或使血清通过柱而被完全吸收。在存在0.2 M NaCl的情况下,不完全抗体的吸收受到抑制;洗脱液组分中仅显示出完整抗体。不完全抗体的洗脱产率较低;在某些情况下,根本无法检测到。这部分可以用非特异性吸收来解释,部分可以用初始抗体滴度较低来解释。因此,用这种方法很难将各个不完全抗体彼此分离。含有红细胞基质的柱仅因其吸收能力(去除不完全抗体)而推荐用于不完全抗体,而不是因其洗脱产率。

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