title: Reducing heterophilic antibody interference in immunoassays using single-chain antibodies.
abstract: Sandwich enzyme-linked immunosorbent assay (ELISA) microarrays can simultaneously quantify the levels of multiple diagnostic targets in a biological sample. However, as with traditional ELISA diagnostics, endogenous antibodies in patient sera can cause interference. We demonstrate here that reducing the diagnostic capture antibody to its minimal functional unit (i.e., a single-chain antibody fragment [scFv]) is an effective strategy for reducing assay interference. Our finding illustrates a source of error introduced by the reliance on immunoglobulin-based capture reagents in sandwich immunoassays with human serum samples. We demonstrate that scFvs can be used in such assays to improve reliability by reducing heterophilic antibody interference, thereby improving biomarker analysis and validation.
authors.affiliation: Pacific Northwest National Laboratory, Richland, WA 99352, USA. cheryl.baird@pnl.gov
authors.forename: Cheryl L
authors.initials: CL
authors.lastname: Baird
authors.forename: Ruimin
authors.initials: R
authors.lastname: Tan
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Fischer
authors.forename: Kristin D
authors.initials: KD
authors.lastname: Victry
authors.forename: Richard C
authors.initials: RC
authors.lastname: Zangar
authors.forename: Karin D
authors.initials: KD
authors.lastname: Rodland
mesh_terms: D000818:Animals; D000910:Antibodies, Heterophile / Q000276:immunology*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D057127:Single-Chain Antibodies / Q000097:blood / Q000276:immunology*
chemical_list: D000910:Antibodies, Heterophile; D007074:Immunoglobulin G; D057127:Single-Chain Antibodies