Adaptation of an ELISA assay for detection of IgG2a responses against therapeutic monoclonal antibodies in a mouse immunization model

Biotherapeutic monoclonal antibodies (mAb) play important roles in clinical medicine but their potential to elicit immune responses in patients remains a major issue. In a study designed to investigate the effect of aggregation on immunogenic responses, mice were immunized with two monoclonal antibo...

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Published in:Journal of immunotoxicology 2022-12, Vol.19 (1), p.1-7
Main Authors: Rane, Shraddha S, Dearman, Rebecca J, Kimber, Ian, Derrick, Jeremy P
Format: Article
Language:English
Subjects:
ADA
Aggregates
aggregation
ELISA
Enzyme-linked immunosorbent assay
Experiments
IgG
igg2a
Immune response
Immunization
Immunogenicity
immunoglobulin
Immunoglobulin G
Monoclonal antibodies
monoclonal antibody
Proteins
Reagents
Regulatory approval
secondary antibody
Serum levels
Software
Variance analysis
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container_title Journal of immunotoxicology
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creator Rane, Shraddha S
Dearman, Rebecca J
Kimber, Ian
Derrick, Jeremy P
description Biotherapeutic monoclonal antibodies (mAb) play important roles in clinical medicine but their potential to elicit immune responses in patients remains a major issue. In a study designed to investigate the effect of aggregation on immunogenic responses, mice were immunized with two monoclonal antibodies (mAb1 and mAb2). Serum levels of total IgG, IgG 1 , and IgG 2a were measured by ELISA. An anti-mouse IgG 2a monoclonal detection antibody cross-reacted with mAb2 but not mAb1, leading to high background when the ELISA plate was coated with mAb2. The problem was solved by use of a goat anti-mouse IgG 2a polyclonal antibody that demonstrated the required specificity. IgG 2a responses were similar for monomer- or aggregate-coated ELISA plates. The results demonstrate the importance of assessment of the specificity of individual reagents when measuring antibody responses against therapeutic antibodies by ELISA.
doi_str_mv 10.1080/1547691X.2021.2020937
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source Taylor & Francis Open Access; Publicly Available Content Database
subjects ADA
Aggregates
aggregation
ELISA
Enzyme-linked immunosorbent assay
Experiments
IgG
igg2a
Immune response
Immunization
Immunogenicity
immunoglobulin
Immunoglobulin G
Monoclonal antibodies
monoclonal antibody
Proteins
Reagents
Regulatory approval
secondary antibody
Serum levels
Software
Variance analysis
title Adaptation of an ELISA assay for detection of IgG2a responses against therapeutic monoclonal antibodies in a mouse immunization model
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