Design of a single-step immunoassay principle based on the combination of an enzyme-labeled antibody release coating and a hydrogel copolymerized with a fluorescent enzyme substrate in a microfluidic capillary device.
A combination of an enzyme-labeled antibody release coating and a novel fluorescent enzyme substrate-copolymerized hydrogel in a microchannel for a single-step, no-wash microfluidic immunoassay is demonstrated. This hydrogel discriminates the free enzyme-conjugated antibody from an antigen-enzyme-conjugated antibody immunocomplex based on the difference in molecular size. A selective and sensitive immunoassay, with 10-1000 ng mL(-1) linear range, is reported.
Graduate School of Engineering, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai City, Osaka 599-8531, Japan. hisamoto@chem.osakafu-u.ac.jp.
Hideki
H
Wakayama
Terence G
TG
Henares
Kaede
K
Jigawa
Shun-ichi
S
Funano
Kenji
K
Sueyoshi
Tatsuro
T
Endo
Hideaki
H
Hisamoto
D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism; D000906:Antibodies / Q000737:chemistry / Q000276:immunology*; D000941:Antigens / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D003374:Coumarins / Q000737:chemistry; D004798:Enzymes / Q000737:chemistry* / Q000378:metabolism; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D058105:Polymerization; D013379:Substrate Specificity
D000906:Antibodies; D000941:Antigens; D003374:Coumarins; D004798:Enzymes; D004800:Enzymes, Immobilized; D005456:Fluorescent Dyes; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D000469:Alkaline Phosphatase