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Microchip immunoaffinity electrophoresis of antibody–thymidine kinase 1 complex
Author(s) -
Pagaduan Jayson V.,
Ramsden Madison,
O'Neill Kim,
Woolley Adam T.
Publication year - 2015
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201400436
Subject(s) - gel electrophoresis , antibody , recombinant dna , microbiology and biotechnology , electrophoresis , chromatography , materials science , biology , chemistry , biochemistry , immunology , gene
Thymidine kinase 1 (TK1) is an important cancer biomarker whose serum levels are elevated in early cancer development. We developed a microchip electrophoresis immunoaffinity assay to measure recombinant purified TK1 (pTK1) using an antibody (Ab) that binds to human TK1. We fabricated PMMA microfluidic devices to test the feasibility of detecting Ab–pTK1 immune complexes as a step toward TK1 analysis in clinical serum samples. We were able to separate immune complexes from unbound Abs using 0.5× PBS (pH 7.4) containing 0.01% Tween‐20, with 1% w/v methylcellulose that acts as a dynamic surface coating and sieving matrix. Separation of the Ab and Ab–pTK1 complex was observed within a 5 mm effective separation length. This method of detecting pTK1 is easy to perform, requires only a 10 μL sample volume, and takes just 1 min for separation.