Reducing interference from heterophilic antibodies in a two-site immunoassay for carcinoembryonic antigen (CEA) by using a human/mouse chimeric antibody to CEA as the tracer

Kuroki, Masahide; Matsumoto, Y.; Arakawa, Fumiko; Haruno, Masatora; Murakami, Masaaki; Kuwahara, Maki; Ozaki, Hiroaki; Senba, Tarumi; Matsuoka, Yuji

doi:10.1016/0022-1759(94)00301-c

Cited by 43 publications

(11 citation statements)

References 32 publications

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“…In this regard, the development of techniques to genetically engineer and produce recombinant antibodies has permitted their evaluation as alternative reagents in diagnostic immunoassays. A human/mouse chimeric antibody has been used previously as a tracer in a two-site enzyme immunoassay for CEA (34 ). This reagent was produced by fusion of V H and V L genes from a murine hybridoma to the constant region heavy and light chain genes derived from a human myeloma cell line.…”

Section: Discussionmentioning

confidence: 99%

Use of an In Vivo Biotinylated Single-Chain Antibody as Capture Reagent in an Immunometric Assay to Decrease the Incidence of Interference from Heterophilic Antibodies

Warren¹,

Bjerner

Paus

et al. 2005

Clinical Chemistry

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Background: Heterophilic antibodies are a common source of interference in immunometric assays. We tested the hypothesis that the incidence of such interference could be decreased by use of a recombinant in vivo-biotinylated single-chain antibody (scFv) as the capture reagent. Methods: We established three assays for carcinoembryonic antigen (CEA) with the capture antibody either chemically biotinylated whole monoclonal T84.66 immunoglobulin, a corresponding F(ab) 2 fragment, or a site-specifically biotinylated T84.66-derived singlechain antibody (scFv). Antibodies were attached to streptavidin-coated microplates. A common europiumlabeled anti-CEA tracer monoclonal antibody was used. The F(ab) 2 assay used a buffer that contained bovine immunoglobulin and aggregated irrelevant monoclonal antibody MAK33 as blocking agents. The whole T84.66 immunoglobulin and scFv assays were performed without addition of blocking agents. From a previous study of 11 261 sera, we tested 390 samples that had displayed heterophilic antibody interference and 179 samples that had not. Results: After correction for bias and analytical variation [2.56 ؋ SD (from the precision profile)], 383 samples displayed significantly different values (>1 g/L) in the whole T84.66-based assay and the F(ab) 2 assay. In contrast, only nine samples showed falsely high CEA concentrations in the scFv assay. After blocking agents were added to the assay buffer, eight of the nine samples displayed results equivalent to those of the

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Section: Discussionmentioning

confidence: 99%

Use of an In Vivo Biotinylated Single-Chain Antibody as Capture Reagent in an Immunometric Assay to Decrease the Incidence of Interference from Heterophilic Antibodies

Warren¹,

Bjerner

Paus

et al. 2005

Clinical Chemistry

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“…The use of a human-mouse chimeric labelled antibody has been shown to be more effective than addition of normal mouse serum in reducing interference in a two-site assay for CEA. 73 In view of the potential seriousness and increasing frequency of interference from HAMA, methods have been developed to measure its concentration in…”

Section: Hctcrophilic Antibodiesmentioning

confidence: 99%

Interference in Immunoassay

1999

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Additional key phrases: heterophilic antibodies; hook effects; matrix effects; specificity 'reagent excess' (type I) and 'reagent limited' (type II) assays. IMMUNOASSAY REACTIONS 'd~':rain ",stionFabfrar ;;Anti body binding site N pepsii~n / digest/-F("')'f'F IGURE I. Stylized diagram of the immunoglobulin molecule showing derivation of the various fragments used for preparation of antibodies in immunoassay.The editor regretfully notes that Mr Colin Selby has died since submitting this review. This article was prepared under the auspices of the Analytical Investigations Standing Committee of the Association of Clinical Biochemists.

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“…Various methods have been proposed to alert laboratories to the presence of or to block the effects of these interfering substances (3,4 ). Antibody microarrays are often 2-site immunoassays configured with antibody pairs, with the 2 antibodies being of mouse origin, rather than a mouse monoclonal antibody and a polyclonal antibody from another species.…”

Section: To the Editormentioning

confidence: 99%

Detection and Elimination of Interference by the Heterophilic Antibody in Antibody Microarray–Based Immunoassay

Yang

Lin

et al. 2011

Clinical Chemistry

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Reducing interference from heterophilic antibodies in a two-site immunoassay for carcinoembryonic antigen (CEA) by using a human/mouse chimeric antibody to CEA as the tracer

Cited by 43 publications

References 32 publications

Use of an In Vivo Biotinylated Single-Chain Antibody as Capture Reagent in an Immunometric Assay to Decrease the Incidence of Interference from Heterophilic Antibodies

Use of an In Vivo Biotinylated Single-Chain Antibody as Capture Reagent in an Immunometric Assay to Decrease the Incidence of Interference from Heterophilic Antibodies

Interference in Immunoassay

Detection and Elimination of Interference by the Heterophilic Antibody in Antibody Microarray–Based Immunoassay

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