化学
糖基化
色谱法
十二烷基硫酸钠
毛细管电泳
质谱法
串联质谱法
加合物
凝胶电泳
生物化学
有机化学
受体
作者
Timothy Kaschak,Daniel Boyd,Boxu Yan
标识
DOI:10.1016/j.ab.2011.06.024
摘要
We report a case study of characterization of a non-enzymatically glycated IgG1 using reducing capillary electrophoresis sodium dodecyl sulfate (CE–SDS) and mass spectrometry (MS). Glycation was found to occur nonspecifically at multiple sites in both the light and heavy chains. The glycated light and heavy chains result in wider peaks eluting late in the reducing CE–SDS profile; in particular, the glycated light chain behaved as a shoulder peak detected by either ultraviolet (UV) or laser-induced fluorescence (LIF) signals. The glycated species can be enriched by boronate affinity chromatography. Analyzing the enriched samples by reversed phase high-performance liquid chromatography in line with time-of-flight MS (RP–HPLC–TOF/MS) revealed adducts of +162 and +324 Da to both the light and heavy chains, suggesting the presence of multiple glycation sites. Tryptic peptide mapping and tandem mass sequencing were used to identify two glycation sites on each of the light and heavy chains.
科研通智能强力驱动
Strongly Powered by AbleSci AI