单体
化学
二聚体
产量(工程)
热稳定性
单一债券
结晶学
立体化学
聚合物
材料科学
有机化学
冶金
烷基
作者
Eve E. Weatherill,Katharine Cain,Sam Heywood,Joanne E. Compson,James T. Heads,Ralph Adams,David P. Humphreys
出处
期刊:Protein Engineering Design & Selection
[Oxford University Press]
日期:2012-05-14
卷期号:25 (7): 321-329
被引量:38
标识
DOI:10.1093/protein/gzs021
摘要
Engineered introduction of interface interchain disulphide bonds is perceived to be a simple method to increase the stability of single chain Fv (scFv). Six disulphide bond locations have been cited within the literature but the potential for the broad use of each has not been examined. Five of these disulphide bond locations were introduced into one scFv in order to compare their relative effects on expression, thermal stability, percent monomer formation and retention of antigen binding. The disulphide bond position vH44–vL100 was observed to enable the most favourable balance of biophysical properties. The vH44–vL100 disulphide bond was introduced into five additional scFv in both vL–vH and vH–vL orientations in order to investigate its general applicability. Data are presented to show the relative influence of scFv sequence, v-region organisation and interchain disulphide bond on expression yield, thermal stability and percent monomer. Introduction of the vH44–vL100 disulphide bond typically resulted in no or little increase in thermal stability and no change in percent monomer but did confer the benefit of permanently fixing monomer:dimer ratios during purification and analysis.
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