Surface modification for enhancing antibody binding on polymer-based microfluidic device for enzyme-linked immunosorbent assay.
A novel surface treatment method using poly(ethyleneimine) (PEI), an amine-bearing polymer, was developed to enhance antibody binding on the poly(methyl methacrylate) (PMMA) microfluidic immunoassay device. By treating the PMMA surface of the microchannel on the microfluidic device with PEI, 10 times more active antibodies can be bound to the microchannel surface as compared to those without treatment or treated with the small amine-bearing molecule, hexamethylenediamine (HMD). Consequently, PEI surface modification greatly improved the immunoassay performance of the microfluidic device, making it more sensitive and reliable in the detection of IgG. The improvement can be attributed to the spacer effect as well as the functional amine groups provided by the polymeric PEI molecules. Due to the smaller dimensions (140x125 microm) of the microchannel, the time required for antibody diffusion and adsorption onto the microchannel surface was reduced to only several minutes, which was 10 times faster than the similar process carried out in 96-well plates. The microchip also had a wider detection dynamic range, from 5 to 1000 ng/mL, as compared to that of the microtiter plate (from 2 to 100 ng/mL). With the PEI surface modification, PMMA-based microchips can be effectively used for enzyme linked immunosorbent assays (ELISA) with a similar detection limit, but much less reagent consumption and shorter assay time as compared to the conventional 96-well plate.
Bai
Yunling
Y
Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio 43210, USA.
Koh
Chee Guan
CG
Boreman
Megan
M
Juang
Yi-Je
YJ
Tang
I-Ching
IC
Lee
L James
LJ
Yang
Shang-Tian
ST
D000818:Animals; D003959:Diamines / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation* / Q000379:methods*; D007074:Immunoglobulin G / Q000737:chemistry* / Q000276:immunology; D007700:Kinetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D018625:Microscopy, Atomic Force; D015394:Molecular Structure; D011094:Polyethyleneimine / Q000737:chemistry*; D019904:Polymethyl Methacrylate / Q000737:chemistry*; D011485:Protein Binding; D051381:Rats; D013057:Spectrum Analysis; D013499:Surface Properties; D014965:X-Rays
D003959:Diamines; D007074:Immunoglobulin G; D011094:Polyethyleneimine; D019904:Polymethyl Methacrylate; C007297:1,6-diaminohexane