title: A receptor-binding-based bioassay to determine the potency of a plasmid biopharmaceutical encoding VEGF-C.
abstract: Over the last decade biological assays (bioassays) have gained much importance for quality control in biopharmaceutical development and manufacturing. Here we describe the development and validation of a bioassay to determine the biological activity (potency) of the plasmid biopharmaceutical pVGI.1 which encodes the VEGF-C (VEGF-2) protein. This assay was developed to test drug substance and drug product for release and stability testing for phase I and II clinical trials. The main focus was on fast assay development and easy handling of the assay, combined with valid results representing the specific therapeutic mechanism. The method includes the expression of the VEGF-C protein in mammalian cells and its binding to the cell surface receptor VEGFR-3. The binding activity of VEGF-C to its immobilized receptor is quantified in a colorimetric assay. IC50 values of VEGF-C expressed after transfection with sample plasmid and an in-house standard plasmid are determined. The ratio of the IC50 value of the test article to that of the reference standard reflects the potency of the sample. The potency assay meets the criteria generally requested by authorities for precision, linearity, accuracy, and range. Therefore the assay can be used in pharmaceutical quality control and is a suitable basis for development of related bioassays.
authors.affiliation: Laboratory of ELISA- and bioassay methods, Department of Quality Control, Boehringer Ingelheim Austria GmbH, Dr. Boehringer-Gasse 5-11, 1121, Vienna, Austria. thomas.waerner@vie.boehringer-ingelheim.com
authors.forename: Thomas
authors.initials: T
authors.lastname: Waerner
authors.forename: Thomas
authors.initials: T
authors.lastname: Girsch
authors.forename: Sandra
authors.initials: S
authors.lastname: Varga
authors.forename: Lichun
authors.initials: L
authors.lastname: Huang
authors.forename: Alexander
authors.initials: A
authors.lastname: Gornikiewicz
authors.forename: Gerhard
authors.initials: G
authors.lastname: Loeber
mesh_terms: D001681:Biological Assay / Q000379:methods*; D015195:Drug Design; D005822:Genetic Vectors; D020128:Inhibitory Concentration 50; D004364:Pharmaceutical Preparations / Q000032:analysis*; D010957:Plasmids / Q000235:genetics; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D013997:Time Factors; D042582:Vascular Endothelial Growth Factor C / Q000235:genetics* / Q000378:metabolism*; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000235:genetics / Q000378:metabolism*
chemical_list: D004364:Pharmaceutical Preparations; D042582:Vascular Endothelial Growth Factor C; D040321:Vascular Endothelial Growth Factor Receptor-3