Covert cross reactants in a two-site immunoassay studied with monoclonal antibodies

Lynette M. Boscato; Gregory Egan; Margaret C. Stuart

doi:10.1016/0003-2697(85)90557-3

Covert cross reactants in a two-site immunoassay studied with monoclonal antibodies

Lynette M. Boscato^*, Gregory Egan, Margaret C. Stuart

^*Corresponding author for this work

Deputy Vice-Chancellor (Research) - Office

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

A one-step two-site immunoradiometric assay for the measurement of free β subunit of human chorionic gonadotrophin (β-hCG) was developed using monoclonal antibodies. The immobilized antibody was specific for free β subunit and the radiolabeled antibody recognized both intact human chorionic gonadotrophin (hCG) and free β subunit. Although the level of hCG "cross-reaction" was low when studied using conventional techniques, the apparent β-hCG content of samples was found to be inversely proportional to the hCG level. From both experimental evidence and computer simulation studies this was found to be due to the binding of hCG to the limited amount of ¹²⁵I-labeled antibody present. The term covert cross reactants has been introduced to describe substances which bind to only one of the antibodies in a two-site immunoassay. When establishing such an assay the effect of covert cross reactants on the response of an analyte should be investigated.

Original language	English
Pages (from-to)	393-401
Number of pages	9
Journal	Analytical Biochemistry
Volume	146
Issue number	2
DOIs	https://doi.org/10.1016/0003-2697(85)90557-3
Publication status	Published - 1 May 1985

Keywords

binding assays
clinical chemistry
computer methods
immunochemical methods
monoclonal antibodies
protein hormones

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10.1016/0003-2697(85)90557-3

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title = "Covert cross reactants in a two-site immunoassay studied with monoclonal antibodies",

abstract = "A one-step two-site immunoradiometric assay for the measurement of free β subunit of human chorionic gonadotrophin (β-hCG) was developed using monoclonal antibodies. The immobilized antibody was specific for free β subunit and the radiolabeled antibody recognized both intact human chorionic gonadotrophin (hCG) and free β subunit. Although the level of hCG {"}cross-reaction{"} was low when studied using conventional techniques, the apparent β-hCG content of samples was found to be inversely proportional to the hCG level. From both experimental evidence and computer simulation studies this was found to be due to the binding of hCG to the limited amount of 125I-labeled antibody present. The term covert cross reactants has been introduced to describe substances which bind to only one of the antibodies in a two-site immunoassay. When establishing such an assay the effect of covert cross reactants on the response of an analyte should be investigated.",

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AU - Boscato, Lynette M.

AU - Egan, Gregory

AU - Stuart, Margaret C.

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Y1 - 1985/5/1

N2 - A one-step two-site immunoradiometric assay for the measurement of free β subunit of human chorionic gonadotrophin (β-hCG) was developed using monoclonal antibodies. The immobilized antibody was specific for free β subunit and the radiolabeled antibody recognized both intact human chorionic gonadotrophin (hCG) and free β subunit. Although the level of hCG "cross-reaction" was low when studied using conventional techniques, the apparent β-hCG content of samples was found to be inversely proportional to the hCG level. From both experimental evidence and computer simulation studies this was found to be due to the binding of hCG to the limited amount of 125I-labeled antibody present. The term covert cross reactants has been introduced to describe substances which bind to only one of the antibodies in a two-site immunoassay. When establishing such an assay the effect of covert cross reactants on the response of an analyte should be investigated.

AB - A one-step two-site immunoradiometric assay for the measurement of free β subunit of human chorionic gonadotrophin (β-hCG) was developed using monoclonal antibodies. The immobilized antibody was specific for free β subunit and the radiolabeled antibody recognized both intact human chorionic gonadotrophin (hCG) and free β subunit. Although the level of hCG "cross-reaction" was low when studied using conventional techniques, the apparent β-hCG content of samples was found to be inversely proportional to the hCG level. From both experimental evidence and computer simulation studies this was found to be due to the binding of hCG to the limited amount of 125I-labeled antibody present. The term covert cross reactants has been introduced to describe substances which bind to only one of the antibodies in a two-site immunoassay. When establishing such an assay the effect of covert cross reactants on the response of an analyte should be investigated.

KW - binding assays

KW - clinical chemistry

KW - computer methods

KW - immunochemical methods

KW - monoclonal antibodies

KW - protein hormones

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Covert cross reactants in a two-site immunoassay studied with monoclonal antibodies

Abstract

Keywords

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