Neutralizing Antibody Assay Development with High Drug and Target Tolerance to Support Clinical Development of an Anti-TFPI Therapeutic Monoclonal Antibody
| Autor: | Boris Gorovits, Alison Joyce, Darshana Jani, Katrina Olson, Chuenlei Parng, Elena Seletskaia, Teresa M Caiazzo, Jean Donley, Yuhong Xiang |
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| Rok vydání: | 2018 |
| Předmět: |
Drug
medicine.drug_class media_common.quotation_subject Lipoproteins Pharmaceutical Science Pharmacology Monoclonal antibody Antibodies Monoclonal Humanized Ligands 030226 pharmacology & pharmacy Binding Competitive Efficacy 03 medical and health sciences 0302 clinical medicine Tissue factor pathway inhibitor Drug Development Drug tolerance medicine Immune Tolerance Humans Neutralizing antibody media_common Immunoassay biology business.industry Immunogenicity Drug Tolerance Antibodies Neutralizing Recombinant Proteins 030220 oncology & carcinogenesis biology.protein Biological Assay Antibody business Protein Binding |
| Zdroj: | The AAPS journal. 21(3) |
| ISSN: | 1550-7416 |
| Popis: | Immunogenicity is a major challenge for protein therapeutics which can potentially reduce drug efficacy and safety and is often being monitored by anti-drug antibody (ADA) and neutralizing antibody (NAb) assays. Circulating targets and residual drugs in matrices can have significant impacts on accuracy of results from ADA and NAb assays, and sufficient drug and target tolerance for these assays are necessary. Here, we report the development of a competitive ligand binding (CLB) NAb assay for an anti-TFPI (tissue factor pathway inhibitor) monoclonal antibody (PF-06741086) with high drug and target tolerance to support ongoing clinical studies. A double acid affinity capture elution approach was used to mitigate drug interference, and a robust target removal strategy was employed to enhance target tolerance. The validated NAb assay has sensitivity of 313 ng/mL, drug tolerance of 50 μg/mL, and target tolerance of 1200 ng/mL. A step-by-step tutorial of assay development is described in this manuscript along with the rationale for using a high drug/target tolerant NAb assay. The NAb assay cut point factor obtained was 0.78. Other assay performance characteristics, e.g., precision and selectivity, are also discussed. This validated method demonstrated a superior drug and target tolerance to warrant specific and precise characterization of the NAb responses in support of ongoing clinical studies. |
| Databáze: | OpenAIRE |
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