An ELISA plate-based assay for hyaluronan using biotinylated proteoglycan G1 domain (HA-binding region).
An enzyme-linked absorbent assay for the determination of hyaluronan (HA) has been developed. The procedure is sensitive, simple and is based on a microtitre plate format. The assay involves competition between HA absorbed to the plate and HA free in solution for binding to biotinylated cartilage proteoglycan binding region (G1 domain). The range of the assay is 10-2500 ng/ml with 50% inhibition at about 200 ng/ml. The assay can be used in guanidine HCl up to 0.6 M and in 0.5% deoxycholate or 0.5% nonidet. This new technique involves fewer experimental steps and is simpler to perform than other methods.
Fosang
A J
AJ
Kennedy Institute of Rheumatology, Hammersmith, London, England.
Hey
N J
NJ
Carney
S L
SL
Hardingham
T E
TE
D000818:Animals; D001710:Biotin; D002356:Cartilage / Q000737:chemistry; D003902:Detergents / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D006025:Glycosaminoglycans / Q000494:pharmacology; D019791:Guanidine; D006146:Guanidines / Q000494:pharmacology; D006801:Humans; D006820:Hyaluronic Acid / Q000032:analysis*; D008832:Microchemistry; D008970:Molecular Weight; D011509:Proteoglycans / Q000302:isolation & purification; D013552:Swine
D003902:Detergents; D006025:Glycosaminoglycans; D006146:Guanidines; D011509:Proteoglycans; D001710:Biotin; D006820:Hyaluronic Acid; D019791:Guanidine