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Protein Microarray Copying: Easy on‐Demand Protein Microarray Generation Compatible with Fluorescence and Label‐Free Real‐Time Analysis
Author(s) -
Kilb Normann,
Herz Tobias,
Burger Jürgen,
Woehrle Johannes,
Meyer Philipp A.,
Roth Günter
Publication year - 2019
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201800699
Subject(s) - protein microarray , microarray , dna microarray , protein array analysis , multiplex , nitrilotriacetic acid , microarray analysis techniques , chemistry , microbiology and biotechnology , computational biology , biology , biochemistry , gene , bioinformatics , gene expression , organic chemistry , chelation
Protein microarrays are essential to understand complex protein interaction networks. Their production, however, is a challenge and renders this technology unattractive for many laboratories. Recent developments in cell‐free protein microarray generation offer new opportunities, but are still expensive and cumbersome in practice. Herein, we describe a cost‐effective and user‐friendly method for the cell‐free production of protein microarrays. From a polydimethylsiloxane (PDMS) flow cell containing an expressible DNA microarray, proteins of interest are synthesised by cell‐free expression and then immobilised on a capture surface. The resulting protein microarray can be regarded as a “copy” of the DNA microarray. 2 His 6 ‐ and Halo‐tagged fluorescent reference proteins were used to demonstrate the functionality of nickel nitrilotriacetic acid (Ni‐NTA) and Halo‐bind surfaces in this copy system. The described process can be repeated several times on the same DNA microarray. The identity and functionality of the proteins were proven during the copy process by their fluorescence and on the surface through a fluorescent immune assay. Also, single‐colour reflectometry (SCORE) was applied to show that, on such copied arrays, real‐time binding kinetic measurements were possible.