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[牛γ球蛋白与氯甲酸酯活化纤维素珠、溴化氰活化琼脂糖凝胶CL-4B及高碘酸钠氧化琼脂糖凝胶6B的偶联及其偶联产物在免疫亲和色谱中的应用]

[Coupling of bovine gamma globulin to carbonochloridate-activated cellulose beads, CNBr-activated Sepharose CL-4B and NaIO4-oxidized Sepharose 6B and use of the coupled products for immunoaffinity chromatography].

作者信息

Hädge D

机构信息

Bereich Tierphysiologie und Immunbiologie, Karl-Marx-Universität Leipzig.

出版信息

Allerg Immunol (Leipz). 1990;36(4):299-307.

PMID:1711284
Abstract

For coupling 25 mg of bovine IgG (BGG) were given to 5 ml volumes of packed bead cellulose activated by 5-norbornene-2.3-dicarboximido carbonochloridate and CNBr-activated Sepharose CL-4B, respectively. Thus, BGG-immunosorbents were obtained with 4.6 to 4.9 mg BGG/ml matrix. 5 ml volumes of packed NaIO4-oxidized Sepharose 6B coupled 27% from 25 mg of BGG, only. In this case, immunosorbents with 1.35 mg BGG/ml matrix were produced. All BGG-immunosorbents were chemically relatively stable. The use of these immunosorbents for affinity chromatography results in the isolation of one milligram of pure rabbit anti-BGG antibodies by means of about 4.6 mg of BGG coupled to the cellulose or the Sepharose-CL-4B matrices. On the other side, only 3.4 mg of BGG coupled to the NaIO4-activated Sepharose 6B were necessary in order to isolate one milligram of antibodies in an immunoelectrophoretically pure state.

摘要

为了进行偶联,分别向5毫升经5-降冰片烯-2,3-二羧亚胺基碳酰氯活化的填充珠状纤维素和CNBr活化的琼脂糖凝胶CL-4B中加入25毫克牛IgG(BGG)。由此获得了BGG免疫吸附剂,每毫升基质含有4.6至4.9毫克BGG。5毫升经高碘酸钠氧化的琼脂糖凝胶6B仅偶联了25毫克BGG中的27%。在这种情况下,制备出了每毫升基质含有1.35毫克BGG的免疫吸附剂。所有BGG免疫吸附剂在化学性质上相对稳定。将这些免疫吸附剂用于亲和色谱法时,通过约4.6毫克偶联到纤维素或琼脂糖凝胶CL-4B基质上的BGG可分离出1毫克纯兔抗BGG抗体。另一方面,为了以免疫电泳纯的状态分离出1毫克抗体,仅需要3.4毫克偶联到高碘酸钠活化的琼脂糖凝胶6B上的BGG。

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