Signal amplification of target protein on heparin glycan microarray.
A heparin glycan chip (HepGlyChip) with a 4800-fold enhanced signal-to-noise ratio as compared with the control without heparin was developed for high-throughput analysis of heparin-protein interactions for new drug development and for screening biological samples in diagnostic applications. As a proof of concept, a heparin glycan microarray was prepared on a poly(styrene-co-maleic anhydride) (PS-MA)-coated glass slide. Heparin was covalently immobilized on poly-l-lysine (PLL) layer with multiple binding sites by sulfo-ethylene glycol bis(succinimidylsuccinate) (sulfo-EGS), increasing the signal-to-noise ratio, minimizing nonspecific binding of target proteins, and resulting in a three-dimensional (3D) structure on the HepGlyChip. This on-chip signal amplification platform was successfully demonstrated by probing the heparin microarray with the highly specific heparin-binding protein antithrombin III (AT III).
Park
Tae-Joon
TJ
Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
Lee
Moo-Yeal
MY
Dordick
Jonathan S
JS
Linhardt
Robert J
RJ
D006493:Heparin / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*
D011134:Polysaccharides; D006493:Heparin