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抗对氨基苯甲酸酯抗体轻链可变区的部分序列:一个单溶剂测序仪程序

Partial sequence of the variable region of an anti-p-azobenzoate antibody light chain: a one solvent sequenator program.

作者信息

Friedenson B, Hora J, Wang L S

出版信息

Biochemistry. 1976 Nov 2;15(22):4876-80. doi: 10.1021/bi00667a019.

Abstract

The partial amino acid sequence of the variable region of a rabbit anti-p-azobenzoate antibody was determined. The sequence determination was facilitated by a modified dimethylallylamine sequencer program in which 1-chlorobutane is used as both the S1 and S3 solvents. To establish the primary structure of virtually the entire light-chain variable region, two different sequencer runs were sufficient: a double cleavage run of 66 cycles on the intact light chain and a single cleavage run of 40 cycles on a peptide isolated from a tryptic digest of the succinylated light chain. Within the limits of the methods used, the light chain appears completely homogeneous. The heavy chains could be more heterogeneous, since the antibody preparation gives at least 18 bands on isoelectric focusing.

摘要

测定了兔抗对氨基苯甲酸抗体可变区的部分氨基酸序列。序列测定借助改良的二甲基烯丙胺测序程序得以简化,该程序中使用1 - 氯丁烷作为S1和S3溶剂。为确定几乎整个轻链可变区的一级结构,进行两次不同的测序运行就足够了:对完整轻链进行66个循环的双切割运行,以及对从琥珀酰化轻链的胰蛋白酶消化物中分离出的肽段进行40个循环的单切割运行。在所使用方法的限度内,轻链似乎完全均一。重链可能更具异质性,因为该抗体制剂在等电聚焦时至少出现18条带。

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