Facile detection of proteins on a solid-phase membrane by direct binding of dextran-based luminol-biotin chemiluminescent polymer.
Facile and non-radioactive methods are desired for the sensitive detection and quantification of various proteins. Herein we describe a novel chemiluminescence (CL)-detection method of particular proteins based on direct binding of a dextran-luminol-biotin (DLB) CL polymer to the proteins on a poly(vinylidene difluoride) membrane. Among 32 kinds of the proteins screened, several proteins such as drug-metabolizing enzymes, cytochrome p450 (CYP)1A2, CYP2E1, and CYP3A4 had the ability to bind directly to the DLB polymer. The binding site in the polymer was owing to the framework of the modified dextran, which underwent oxidation and reduction procedures. This interaction might be the comprehensive effect of both electrostatic interaction and steric complementarities. CL intensity of the proteins detected by the polymer could be further enlarged by the mediation of avidin. The proposed CL-imaging method possesses potential as a rapid, facile, inexpensive and selective detection of the proteins.
Zhang
Huan
H
Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, 1-14, Bunkyo-Machi, Nagasaki 852-8521, Japan.
Shibata
Takayuki
T
Krawczyk
Tomasz
T
Kabashima
Tsutomu
T
Lu
Jianzhong
J
Lee
Myung K
MK
Kai
Masaaki
M
D000818:Animals; D001360:Avidin / Q000378:metabolism; D001710:Biotin / Q000737:chemistry*; D002417:Cattle; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D003911:Dextrans / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay; D008163:Luminescent Measurements; D008165:Luminol / Q000737:chemistry*; D008567:Membranes, Artificial; D051379:Mice; D011145:Polyvinyls / Q000737:chemistry*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D005465:Fluorocarbon Polymers
D001360:Avidin; D001710:Biotin; D003577:Cytochrome P-450 Enzyme System; D003911:Dextrans; D008165:Luminol; D008567:Membranes, Artificial; D011145:Polyvinyls; D011506:Proteins; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers