Structural diversity among anti-p-azophenylarsonate monoclonal antibodies from A/J mice; comparison of Id- and Id+ sequences.
Structural diversity among anti-p-azophenylarsonate monoclonal antibodies from A/J mice; comparison of Id- and Id+ sequences.
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A/J 小鼠抗对偶氮苯胂酸单克隆抗体的结构多样性；
DOI：
10.1016/0161-5890(81)90022-5
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发表时间：
1981
影响因子：
3.6
通讯作者：
Gefter,ML
中科院分区：
文献类型：
--
作者：
Margolies,MN;Marshak-Rothstein,A;Gefter,ML
对从携带从A/J脾淋巴细胞与骨髓瘤细胞系Sp2/0-Ag 14融合获得的细胞系的小鼠的腹水分离的单克隆抗-对-偶氮苯胂酸盐抗体进行氨基酸序列分析。将来自7个独特型阴性杂交瘤蛋白的重链和轻链的部分一级结构与6个独特型阳性分子的部分一级结构进行比较。来自携带主要交叉反应独特型IdCR的分子的重链的氨基末端氨基酸序列（40-47个残基）彼此具有95%的同源性。类似地，IdCR+轻链的氨基末端序列彼此同源。然而，序列变异在单个抗体的框架和互补决定区中是明显的，这表明一个大的分子家族解释了主要的交叉反应独特型，如先前报道的（Marshak-Rothsteinet等人，对来自七种IdCR阴性单克隆抗体的重链和轻链进行氨基末端（37-48个残基）氨基酸序列分析。四条重链被封闭以防止Edman降解，但可以在酶促去除氨基末端吡咯烷酮羧酸残基后进行测序。与IdCR阳性重链相比，IdCR阴性重链在框架区和互补决定区中表现出更大的多样性，与来自合并的血清IdCR阳性抗体的结果相反，存在几个不同的亚组（Capraet al.，1975年）。七条IdCR阴性轻链中的一条被阻断。其余IdCR阴性轻链的序列在框架区和互补决定区均表现出显著变化，IdCR阳性和IdCR阴性单克隆抗体的氨基末端序列之间的比较表明，在第一互补决定区中的特定序列-重链和轻链的决定区不足以解释主要的交叉反应独特型。A/J小鼠中IdCR的结构基础可能在可变区的其他区段中。
Amino acid sequence analyses were carried out on monoclonal anti-p-azophenylarsonate antibodies isolated from the ascites of mice carrying cell lines obtained from the fusion of A/J splenic lymphocytes with the myeloma cell line Sp2/0–Ag14. The partial primary structures of both heavy and light chains from seven idiotype negative hybridoma proteins are compared to those of six idiotype positive molecules. Amino-terminal amino acid sequences (40–47 residues) of heavy chains from molecules bearing the major cross-reacting idiotype, IdCR, demonstrated 95% homology to each other. Similarly, aminoterminal sequences of IdCR+light chains were homologous to each other. However, sequence variations were evident in individual antibodies in both framework and complementarity-determining regions, suggesting that a large family of molecules accounts for the major cross-reacting idiotype, as previously reported (Marshak-Rothsteinet al., 1980b).Heavy and light chains from seven IdCR-negative monoclonal antibodies were subjected to amino-terminal (37–48 residues) amino acid sequence analysis. Four heavy chains were blocked to Edman degradation, but could be sequenced after enzymatic removal of the amino-terminal pyrrolidone carboxylic acid residue. In comparison with IdCR-positive heavy chains, the IdCR-negative heavy chains demonstrate greater diversity in both framework and complementarity-determining regions, with several different subgroups represented in contrast to the results from pooled serum IdCR-positive antibodies (Capraet al., 1975). One of the seven IdCR-negative light chains was blocked. The sequences of the remaining IdCR-negative light chains exhibited marked variations in both framework and complementarity-determining regions, with different chain lengths in the first complementarity-determining region in several light chains.Comparisons between the amino-terminal sequences of IdCR-positive and IdCR-negative monoclonal antibodies suggest that specific sequences in the first complementarity-determining regions of both heavy and light chains are not sufficient to account for the major cross-reacting idiotype. The structural basis for IdCRin A/J mice is likely to be in other segments of the variable regions.