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

Shraddha Rane, Rebecca Dearman, Ian Kimber, Jeremy Derrick

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Abstract

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, IgG1, and IgG2a were measured by ELISA. An anti-mouse IgG2a 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 IgG2a polyclonal antibody that demonstrated the required specificity. IgG2a 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.
Original languageEnglish
Pages (from-to)1-7
Number of pages7
JournalJournal of immunotoxicology
Volume19
Issue number1
DOIs
Publication statusPublished - 25 Jan 2022

Keywords

  • ADA
  • ELISA
  • IgG
  • aggregation
  • immunogenicity
  • immunoglobulin
  • monoclonal antibody
  • secondary antibody

Research Beacons, Institutes and Platforms

  • Lydia Becker Institute
  • Manchester Institute for Collaborative Research on Ageing

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