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[利用免疫活性合成肽制备抗人乙酰胆碱烟碱受体α7亚基的抗体]

[Production of antibodies to the alpha7-subunit of human acetylcholine nicotinic receptor with the use of immunoactive synthetic peptides].

作者信息

Vol'pina O M, Titova M A, Koroev D O, Volkova T D, Oboznaia M B, Zhmak M N, Alekseev T A, Tsetlin V I

出版信息

Bioorg Khim. 2006 Mar-Apr;32(2):169-75.

Abstract

Potential B epitopes and T-helper epitopes in the N-terminal extracellular domain of the alpha7-subunit of human acetylcholine receptor (AChR) were theoretically calculated in order to reveal peptides that can induce the formation of specific antibodies to this domain. Four peptides structurally corresponding to four alpha7-subunit regions containing 16-23 aa and three of their truncated analogues were synthesized. Rabbits were immunized with both free peptides and protein conjugates of their truncated analogues, and a panel of antibodies to various exposed regions of the N-terminal extracellular domain of the AChR alpha7-subunit was obtained. All of the four predicted peptides were shown to induce the production of antipeptide antibodies in free form, without conjugation with any protein carrier. The free peptides and the protein conjugates of truncated analogues induced the formation of almost equal levels of antibodies. Most of the obtained antisera contained antibodies that bind to the recombinant extracellular N-terminal domain of the rat AChR alpha7-subunit and do not react with the analogous domain of the alpha1-subunit of the ray Torpedo californica AChR.

摘要

为了揭示能够诱导针对人乙酰胆碱受体(AChR)α7亚基N端胞外域产生特异性抗体的肽段,对该区域的潜在B细胞表位和T辅助细胞表位进行了理论计算。合成了四种与包含16 - 23个氨基酸的α7亚基四个区域结构相对应的肽段及其三种截短类似物。用游离肽段及其截短类似物的蛋白偶联物对兔子进行免疫,获得了一组针对AChRα7亚基N端胞外域不同暴露区域的抗体。结果表明,所有四种预测肽段在未与任何蛋白质载体偶联的游离形式下均能诱导抗肽抗体的产生。游离肽段和截短类似物的蛋白偶联物诱导产生的抗体水平几乎相等。所获得的大多数抗血清中含有的抗体能与大鼠AChRα7亚基的重组胞外N端域结合,但不与加州电鳐AChRα1亚基的类似域发生反应。

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