Davies J, Riechmann L
MRC Laboratory of Molecular Biology, Cambridge, UK.
FEBS Lett. 1994 Feb 21;339(3):285-90. doi: 10.1016/0014-5793(94)80432-x.
A human heavy chain variable domain (VH) was expressed in bacteria for structural analysis by NMR spectroscopy. NMR analysis was initially impossible due to the short transverse proton relaxation time of the VH, probably caused by aggregation through the exposed interface naturally in contact with the light chain. The relaxation time was improved to normal values when this interface was mutated to mimic heavy chains of camel antibodies naturally devoid of light chains and through the use of the detergent CHAPS. Assignment of NMR signals will now be possible after isotopic labeling. Implications for the design of VH domains as minimum size immunoreagents are outlined.
一条人源重链可变区（VH）在细菌中表达，用于通过核磁共振波谱进行结构分析。由于VH的横向质子弛豫时间短，核磁共振分析最初无法进行，这可能是由于通过自然与轻链接触的暴露界面发生聚集所致。当该界面发生突变以模拟天然不含轻链的骆驼抗体的重链，并使用去污剂CHAPS时，弛豫时间提高到正常水平。同位素标记后，现在可以对核磁共振信号进行归属。概述了将VH结构域设计为最小尺寸免疫试剂的意义。
