用于核磁共振研究的12种抗二硝基苯基自旋标记单克隆抗体的序列。
Sequences of 12 monoclonal anti-dinitrophenyl spin-label antibodies for NMR studies.
作者信息
Leahy D J, Rule G S, Whittaker M M, McConnell H M
机构信息
Stauffer Laboratory for Physical Chemistry, Stanford University, CA 94305.
出版信息
Proc Natl Acad Sci U S A. 1988 Jun;85(11):3661-5. doi: 10.1073/pnas.85.11.3661.
Eleven monoclonal antibodies specific for a spin-labeled dinitrophenyl hapten (DNP-SL) have been produced for use in NMR studies. They have been named AN01 and AN03-AN12. The stability constants for the association of these antibodies with DNP-SL and related haptens were measured by fluorescence quenching and ranged from 5.0 X 10(4) M-1 to greater than 1.0 X 10(8) M-1. cDNA clones coding for the heavy and light chains of each antibody and of an additional anti-DNP-SL monoclonal antibody, AN02, have been isolated. The nucleic acid sequence of the 5' end of each clone has been determined, and the amino acid sequence of the variable regions of each antibody has been deduced from the cDNA sequence. The sequences are relatively heterogeneous, but both the heavy and the light chains of AN01 and AN03 are derived from the same variable-region gene families as those of the AN02 antibody. AN07 has a heavy chain that is related to that of AN02, and AN09 has a related light chain. AN05 and AN06 are unrelated to AN02 but share virtually identical heavy and light chains. Preliminary NMR difference spectra comparing related antibodies show that sequence-specific assignment of resonances is possible. Such spectra also provide a measure of structural relatedness.
已制备出11种针对自旋标记二硝基苯基半抗原（DNP-SL）的单克隆抗体，用于核磁共振（NMR）研究。它们被命名为AN01和AN03 - AN12。通过荧光猝灭法测定了这些抗体与DNP-SL及相关半抗原结合的稳定常数，范围从5.0×10⁴ M⁻¹到大于1.0×10⁸ M⁻¹。已分离出编码每种抗体以及另一种抗DNP-SL单克隆抗体AN02的重链和轻链的cDNA克隆。已确定每个克隆5'端的核酸序列，并从cDNA序列推导了每种抗体可变区的氨基酸序列。这些序列相对异质，但AN01和AN03的重链和轻链均与AN02抗体来自相同的可变区基因家族。AN07的重链与AN02的重链相关，AN09的轻链相关。AN05和AN06与AN02无关，但重链和轻链几乎相同。比较相关抗体的初步NMR差异光谱表明，共振的序列特异性归属是可能的。此类光谱还提供了结构相关性的一种度量。
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