A simple method to determine IgG light chain to heavy chain polypeptide ratios expressed by CHO cells
Anja Gerster1, Claas Wodarczyk2, Britta Reichenbächer2
1Rentschler Biotechnologie GmbH, Erwin-Rentschler-Straße 21, 88471, Laupheim, Germany. anja.gerster@rentschler.de.
Objectives:
To establish a high-throughput method for determination of antibodies intra- and extracellular light chain (LC) to heavy chain (HC) polypeptide ratio as screening parameter during cell line development.
Results:
Chinese Hamster Ovary (CHO) TurboCell pools containing different designed vectors supposed to result in different LC:HC polypeptide ratios were generated by targeted integration. Cell culture supernatants and cell lysates of a fed batch experiment were purified by combined Protein A and anti-kappa affinity batch purification in 96-well format. Capture of all antibodies and their fragments allowed the determination of the intra- and extracellular LC:HC peptide ratios by reduced SDS capillary electrophoresis. Results demonstrate that the method is suitable to show the significant impact of the vector design on the intra- and extracellular LC:HC polypeptide ratios.
Conclusion:
Determination of LC:HC polypeptide ratios can give important information in vector design optimization leading to CHO cell lines with optimized antibody assembly and preferred product quality.
