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A Direct Immersion System for Peptide Enrichment
Author(s) -
Tai DarFu,
Lin YeeFung,
Lu KuoHao,
Chen GuanYu,
Shu HunChi
Publication year - 2012
Publication title -
journal of the chinese chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.329
H-Index - 45
eISSN - 2192-6549
pISSN - 0009-4536
DOI - 10.1002/jccs.201100635
Subject(s) - chemistry , peptide , chromatography , filter paper , acetonitrile , cellulose , molecular imprinting , extraction (chemistry) , desorption , acetic acid , molecularly imprinted polymer , microtiter plate , adsorption , biochemistry , selectivity , organic chemistry , catalysis
A complementary peptide separation and purification system, using a novel, artificial, affinity‐type extraction material, is described. Cellulose paper was first coated with a 3‐methacryloxypropyltrimethoxysilane (MPS) layer, followed by imprinting with peptides, to form an approximately 2‐μm layer of film (molecularly imprinted polymer film; MIPF) on both sides and resulted in an increased affinity toward the corresponding template as well as to matched polypeptides. MIPF‐cellulose paper (MIFC) was immersed in 2% peptide solution (acetonitrile/water) for the sorption of targeted‐peptides at room temperature for an hour in a 96‐well microtiter plate. Desorption of targeted‐peptides from the MIPF‐ and NIPF (non‐imprinted polymeric film)‐coated filter papers was nearly complete (∼99%) in two desorptions of 10 min using 5% acetic acid in water. Clean extracts of targeted‐peptides were obtained demonstrating the suitability of MIPF‐coated filter papers for the extraction or analysis of biological samples. To compare their extraction characteristics under various conditions, analyses were carried out by an enzyme‐linked immunosorbent assay (ELISA) microplate reader as peptide‐detecting/peptide‐capturing microarrays. In load ability studies using acetonitrile/water, peptide masses up to 100 μg could be applied onto a piece of MIPF coated filter paper (0.7 cm in diameter).