title: Enhanced ELISA based on carboxymethylated dextran coatings.
abstract: In a "sandwich" enzyme-linked immunosorbent assay (ELISA) designed to detect an antigen in a complex protein mixture, the antigen is usually captured via an antibody adsorbed to the wells of a microplate. Plate preparation for standard assay involves a passive adsorption of capture antibodies followed by the incubation of blocking agents. Here, we describe a new strategy that replaces these two time-consuming adsorption steps (up to 15 h) by a unique step corresponding to the covalent grafting of the capture antibody on a carboxymethylated dextran (CMD) layer, a single step completed in 15 min. Taking advantage of the CMD low-fouling properties, blocking agent-free buffer solutions can be used as diluent in the improved approach.
authors.affiliation: Department of Chemical Engineering, École Polytechnique de Montréal, succ. Centre-Ville, 6079, Montréal, QC, Canada, H3C 3A7.
authors.forename: Benoît
authors.initials: B
authors.lastname: Liberelle
authors.forename: Charles
authors.initials: C
authors.lastname: Fortier
authors.forename: Gregory
authors.initials: G
authors.lastname: De Crescenzo
mesh_terms: D000327:Adsorption; D000906:Antibodies / Q000737:chemistry; D000941:Antigens / Q000032:analysis*; D001426:Bacterial Proteins / Q000737:chemistry; D002021:Buffers; D002234:Carbodiimides / Q000737:chemistry; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis; D003911:Dextrans / Q000138:chemical synthesis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D004815:Epidermal Growth Factor / Q000032:analysis*; D006735:Horseradish Peroxidase / Q000737:chemistry; D006801:Humans; D011994:Recombinant Proteins / Q000032:analysis; D012996:Solutions; D013388:Succinimides / Q000737:chemistry
chemical_list: C518491:1-ethyl-3-(3-(diethylamino)propyl)carbodiimide; D000906:Antibodies; D000941:Antigens; D001426:Bacterial Proteins; D002021:Buffers; D002234:Carbodiimides; D020099:Coated Materials, Biocompatible; D003911:Dextrans; D011994:Recombinant Proteins; D012996:Solutions; C058865:Strep-avidin conjugated horseradish peroxidase; D013388:Succinimides; D004815:Epidermal Growth Factor; C014392:carboxymethyl dextran; D006735:Horseradish Peroxidase; C001426:N-hydroxysuccinimide