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Using Protein Microarray Technology to Screen Anti‐PD‐1 Monoclonal Antibodies for Specificity and Applications in Anatomic Pathology
Author(s) -
Chen Caiwei,
Wei Haitao,
Yuan Kehu,
Wu Guiyin,
Chen Jian,
Chu Boyang,
Wang Guangli,
Shu Youmin,
He Weiwu,
Ma Donghui
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.417.9
Subject(s) - monoclonal antibody , immunohistochemistry , pembrolizumab , tissue microarray , companion diagnostic , antibody , monoclonal , protein microarray , medicine , pathology , immunotherapy , targeted therapy , cancer research , melanoma , immune system , cancer , microarray , biology , immunology , biochemistry , gene expression , gene
Programmed death‐1 (PD‐1) expresses in many tumors in response to inflammation. It is up‐regulated in activated T lymphocytes and inhibits T‐cell function upon binding to its ligands PD‐L1 and PDL2 and serves as a key checkpoint of the immune system. KEYTRUDA (pembrolizumab) is the first anti‐PD‐1 therapy approved in the United States and received FDA's Breakthrough Therapy designation for advanced melanoma. Thus, to evaluate PD1 protein levels in formalin‐fixed paraffin‐embedded tissue samples, a high quality monoclonal antibody validated for immunohistochemistry (IHC) is needed. To develop the best monoclonal antibody for PD‐1 IHC analysis, 41 monoclonal antibodies were generated. 8 of them work for IHC application on FFPE tissue sections. To identify clones that are mono‐specific on target, only two clones passed our high density protein microarray chip tests. Other immunoassays and species cross‐reactivity tests were also explored. In summary, two newly generated monoclonal antibodies demonstrated ultra‐specificity against PD‐1 protein and superior performance for IHC analyses. These two clones could be utilized as companion diagnostic reagents to stratify cancer patients before KEYTRUDA prescription. This research is partly supported by Chinese 863 research grant number 2014AA020909.