Validation of an Immunoassay for Anti-thymidine Phosphorylase Antibodies in Patients with MNGIE Treated with Enzyme Replacement Therapy
| Autor: | Jamie Hall, Marcia Sellos-Moura, Charlotte Gasson, Bridget E. Bax, Dario Pacitti, Michelle Levene |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
lcsh:QH426-470 Pharmacology thymidine phosphorylase Article 03 medical and health sciences Glycogen phosphorylase 0302 clinical medicine Drug tolerance Genetics medicine Thymidine phosphorylase lcsh:QH573-671 Molecular Biology medicine.diagnostic_test biology Chemistry lcsh:Cytology Immunogenicity Assay sensitivity Enzyme replacement therapy assay validation 3. Good health bridging immunoassay lcsh:Genetics 030104 developmental biology Immunoassay MNGIE biology.protein Molecular Medicine Antibody 030217 neurology & neurosurgery enzyme replacement therapy |
| Zdroj: | Molecular Therapy: Methods & Clinical Development, Vol 11, Iss, Pp 1-8 (2018) Molecular Therapy. Methods & Clinical Development Molecular Therapy-Methods & Clinical Development |
| ISSN: | 2329-0501 |
| Popis: | Erythrocyte encapsulated thymidine phosphorylase is recombinant Escherichia coli thymidine phosphorylase encapsulated within human autologous erythrocytes and is under development as an enzyme replacement therapy for the ultra-rare inherited metabolic disorder mitochondrial neurogastrointestinal encephalomyopathy. This study describes the method validation of a two-step bridging electrochemiluminescence immunoassay for the detection of anti-thymidine phosphorylase antibodies in human serum according to current industry practice and regulatory guidelines. The analytical method was assessed for screening cut point, specificity, selectivity, precision, prozone effect, drug tolerance, and stability. Key findings were a correction factor of 129 relative light units for the cut-point determination; a specificity cut point of 93% inhibition; confirmed intra-assay and inter-assay precision; assay sensitivity of 356 ng/mL; no matrix or prozone effects up to 25,900 ng/mL; a drug tolerance of 156 ng/mL; and stability at room temperature for 24 hr and up to five freeze-thaws. Immunogenicity evaluations of serum from three patients who received erythrocyte encapsulated thymidine phosphorylase under a compassionate treatment program showed specific anti-thymidine phosphorylase antibodies in one patient. To conclude, a sensitive, specific, and selective immunoassay has been validated for the measurement of anti-thymidine phosphorylase antibodies; this will be utilized in a phase II pivotal clinical trial of erythrocyte encapsulated thymidine phosphorylase. Keywords: assay validation, bridging immunoassay, enzyme replacement therapy, MNGIE, thymidine phosphorylase |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|