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An automated immunoassay for early specificity profiling of antibodies.

Authors
  • Frese, Katrin
  • Eisenmann, Meike
  • Ostendorp, Ralf
  • Brocks, Bodo
  • Pabst, Stefan
Type
Published Article
Journal
mAbs
Publisher
Landes Bioscience
Publication Date
Jan 01, 2013
Volume
5
Issue
2
Pages
279–287
Identifiers
DOI: 10.4161/mabs.23539
PMID: 23412646
Source
Medline
Keywords
License
Unknown

Abstract

Antibody-based therapeutics are of great value for the treatment of human diseases. In addition to functional activity, affinity or physico-chemical properties, antibody specificity is considered to be one of the most crucial attributes for safety and efficacy. Consequently, appropriate studies are required before entering clinical trials. High content protein arrays are widely applied to assess antibody specificity, but this commercial solution can only be applied to final therapeutic antibody candidates because such arrays are expensive and their throughput is limited. A flexible, high-throughput and economical assay that allows specificity testing of IgG or Fab molecules during early discovery is described here. The 384-well microtiter plate assay contains a comprehensive panel of 32 test proteins and uses electrochemiluminescence as readout. The Protein Panel Profiling ( 3P) was used to analyze marketed therapeutic antibodies that all showed highly specific binding profiles. Subsequently, 3P was applied to antibody candidates from early discovery and the results compared well with those obtained with a commercially available high content protein chip. Our results suggest that 3P can be applied as an additional filter for lead selection, allowing the identification of favorable antibody candidates in early discovery and thereby increasing the speed and possibility of success in drug development.

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