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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Bibliographic Details
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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Summary: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.
ISSN:1547-691X
1547-6901
DOI:10.1080/1547691X.2021.2020937