Abstract |
TGN1412 is a "superagonistic" CD28 monoclonal antibody ( IgG4) that caused serious adverse events at its first time in human clinical trial. In the present study, different in vitro methods for detecting and quantifying unwanted pro-inflammatory activity of therapeutic monoclonal antibodies (mAbs) such as TGN1412 are described. The antibody of interest is immobilised by wet-coating or air-drying onto polypropylene or polystyrene 96-well plates prior to the addition of human peripheral blood mononuclear cells (PBMCs). The cells are incubated for 16-24h with the immobilised antibody which allows the accumulation of pro-inflammatory cytokines, quantified by enzyme-linked immunoabsorbent assay (ELISA), in response to the antibody. Cytokine responses stimulated by TGN1412 immobilised by air-drying onto polypropylene and polystyrene plates were much larger than responses to TGN1412 wet-coated onto polypropylene and polystyrene plates, respectively. In additional experiments with other mAbs associated with clinical reactions, air-dried mAbs stimulated larger tumour necrosis factor-alpha (TNF) responses than antibodies added in aqueous phase. Also, TGN1412 air-dried onto plastic plates stimulated large proliferative responses of 3-day cultures of lymphocytes. It was concluded that immobilising mAbs by air-drying offers a useful in vitro method for detecting and quantifying pro-inflammatory activities of therapeutic mAbs.
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Authors | Lucy Findlay, David Eastwood, Richard Stebbings, Giles Sharp, Yogesh Mistry, Christina Ball, John Hood, Robin Thorpe, Stephen Poole |
Journal | Journal of immunological methods
(J Immunol Methods)
Vol. 352
Issue 1-2
Pg. 1-12
(Jan 31 2010)
ISSN: 1872-7905 [Electronic] Netherlands |
PMID | 19895813
(Publication Type: Journal Article)
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Copyright | Copyright 2009 Elsevier B.V. All rights reserved. |
Chemical References |
- Antibodies, Monoclonal
- Antibodies, Monoclonal, Humanized
- CD28 Antigens
- Recombinant Fusion Proteins
- Tumor Necrosis Factor-alpha
- TGN-1412
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Topics |
- Antibodies, Monoclonal
(pharmacology)
- Antibodies, Monoclonal, Humanized
- Binding, Competitive
- CD28 Antigens
(immunology, metabolism)
- Cell Proliferation
(drug effects)
- Cells, Cultured
- Drug Evaluation
(methods)
- Enzyme-Linked Immunosorbent Assay
- Humans
- Immunotherapy, Active
- In Vitro Techniques
- Leukocytes, Mononuclear
(drug effects, immunology)
- Lymphocyte Activation
(drug effects, immunology)
- Recombinant Fusion Proteins
(metabolism)
- Tumor Necrosis Factor-alpha
(metabolism)
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