Frontal affinity chromatography combined on-line with mass spectrometry: a tool for the binding study of different epidermal growth factor receptor inhibitors.
Frontal affinity chromatography (FAC) is a simple but powerful method to analyze molecular interactions between an analyte and an immobilized ligand by calculating the extent of retardation of the elution front. By combination of FAC with a PE-Mariner electrospray ionization mass spectrometry, a very efficient and straightforward procedure was developed herein for analyzing the binding properties of different inhibitors of the epidermal growth factor receptor (EGFR). In this study, a polyclonal antibody prepared with a known anti-EGFR inhibitor coupled with bovine serum albumin was adopted as the stationary phase in the FAC system. Using the antibody to mimic the receptor, other different anti-EGFR inhibitors as well as the small-molecule half-antigen itself were recognized directly from the crude extract of herb, which afforded us a novel promising approach for the efficient screening of lead compounds or drug candidates from natural resources.
Zhu
Lili
L
College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, P. R. China.
Chen
Lirong
L
Luo
Hongpeng
H
Xu
Xiaojie
X
D000818:Animals; D032602:Caragana; D002846:Chromatography, Affinity / Q000379:methods; D066246:ErbB Receptors / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D011817:Rabbits
D010936:Plant Extracts; D066246:ErbB Receptors