title: Selective extraction and enrichment of glycoproteins based on boronate affinity SPME and determination by CIEF-WCID.
abstract: In this study, a new thin-film boronic acid coating was developed for solid-phase microextraction (SPME) followed by capillary isoelectric focusing with whole-column imaging detection (CIEF-WCID). Boronate functionalized particles of phenylboronic acid (PBA) and 3-aminophenylboronic acid (3-aPBA) were utilized as boronate affinity solid phase coating on thin-film stainless steel blades for selective extraction and enrichment of glycoproteins. The process of extraction and elution could be easily controlled by adjusting pH. To test specificity, asialofetuin and lactoferrin were selected as glycoproteins test molecules, while BSA and myoglobin were used as control non-glycoproteins in this study. The boronate affinity coating was characterized. The effect of buffer, pH, extraction profiles and elution profiles were investigated. The developed method was successfully applied to extract glycoproteins from standard buffer, PBS, human plasma and 10-fold diluted human blood using two kinds of boronate affinity blades. Boronate affinity SPME could be a promising tool for selective extraction and enrichment of low-abundance glycoproteins in real biological samples.
authors: Key Laboratory of Oncology, Chongqing Cancer Hospital & Institute & Cancer Center, Chongqing 400030, China; Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment, Chongqing Cancer Institute, Chongqing 400030, China.
authors: Lixian
authors: L
authors: Li
authors: College of Chemistry and Chemical Engineering, Chongqing University, Chongqing 400030, China.
authors: Zhining
authors: Z
authors: Xia
authors: Department of Chemistry, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada. Electronic address: janusz@uwaterloo.ca.
authors: Janusz
authors: J
authors: Pawliszyn
mesh_terms: D001897:Boronic Acids / Q000737:chemistry*; D004867:Equipment Design; D006023:Glycoproteins / Q000032:analysis / Q000097:blood* / Q000302:isolation & purification*; D006801:Humans; D007525:Isoelectric Focusing / Q000295:instrumentation; D052617:Solid Phase Microextraction / Q000295:instrumentation / Q000379:methods*
chemical_list: D001897:Boronic Acids; D006023:Glycoproteins; C028592:3-aminobenzeneboronic acid; C010686:benzeneboronic acid
keywords: Boronate affinity; Capillary isoelectric focusing with whole-column imaging detection; Glycoprotein; Solid phase microextraction