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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adsorption
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Antibodies / chemistry
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Antigens / analysis*
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Bacterial Proteins / chemistry
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Buffers
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Carbodiimides / chemistry
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Coated Materials, Biocompatible / chemical synthesis
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Dextrans / chemical synthesis*
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Enzyme-Linked Immunosorbent Assay / methods*
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Enzyme-Linked Immunosorbent Assay / standards
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Epidermal Growth Factor / analysis*
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Horseradish Peroxidase / chemistry
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Humans
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Recombinant Proteins / analysis
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Solutions
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Succinimides / chemistry
Substances
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1-ethyl-3-(3-(diethylamino)propyl)carbodiimide
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Antibodies
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Antigens
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Bacterial Proteins
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Buffers
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Carbodiimides
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Coated Materials, Biocompatible
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Dextrans
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Recombinant Proteins
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Solutions
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Strep-avidin conjugated horseradish peroxidase
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Succinimides
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Epidermal Growth Factor
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carboxymethyl dextran
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Horseradish Peroxidase
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N-hydroxysuccinimide