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来自肌酸激酶特异性单克隆抗体的重链和轻链cDNA的克隆及核苷酸序列

Cloning and nucleotide sequence of heavy- and light-chain cDNAs from a creatine-kinase-specific monoclonal antibody.

作者信息

Buckel P, Hübner-Parajsz C, Mattes R, Lenz H, Haug H, Beaucamp K

出版信息

Gene. 1987;51(1):13-9. doi: 10.1016/0378-1119(87)90469-0.

Abstract

Determination of creatine kinase isoenzymes by inhibition assay is a useful tool for the diagnosis and monitoring of myocardial infarction. We have established several mouse hybridoma lines secreting monoclonal antibodies with creatine kinase M-subunit inhibitory capacity. One of the monoclonal antibodies (MAK33) inhibits creatine kinase-MM by 80% without influencing the activity of creatine kinase-MB. A combination of two monoclonal antibodies increased the inhibition of creatine kinase MM up to 99.4%. Poly(A) + RNA of hybridoma cells secreting MAK33 was isolated and used for cloning cDNA of both heavy and light chains of this antibody. Full-length cDNA clones were obtained by hybridization with gamma 1 and kappa constant region cDNA probes. The complete nucleotide sequences from the variable regions including signal peptide and part of the 5'-untranslated regions have been determined.

摘要

通过抑制测定法测定肌酸激酶同工酶是诊断和监测心肌梗死的一种有用工具。我们已经建立了几种分泌具有肌酸激酶M亚基抑制能力的单克隆抗体的小鼠杂交瘤细胞系。其中一种单克隆抗体(MAK33)可抑制肌酸激酶-MM达80%,而不影响肌酸激酶-MB的活性。两种单克隆抗体联合使用可将肌酸激酶-MM的抑制率提高至99.4%。分离出分泌MAK33的杂交瘤细胞的聚腺苷酸加尾RNA(Poly(A)+RNA),并用于克隆该抗体重链和轻链的cDNA。通过与γ1和κ恒定区cDNA探针杂交获得了全长cDNA克隆。已经确定了包括信号肽和部分5'非翻译区在内的可变区的完整核苷酸序列。

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