A simple method to determine IgG light chain to heavy chain polypeptide ratios expressed by CHO cells.

Autor: Gerster A; Rentschler Biotechnologie GmbH, Erwin-Rentschler-Straße 21, 88471, Laupheim, Germany. anja.gerster@rentschler.de., Wodarczyk C; Rentschler Biotechnologie GmbH, Erwin-Rentschler-Straße 21, 88471, Laupheim, Germany., Reichenbächer B; Rentschler Biotechnologie GmbH, Erwin-Rentschler-Straße 21, 88471, Laupheim, Germany., Köhler J; Rentschler Biotechnologie GmbH, Erwin-Rentschler-Straße 21, 88471, Laupheim, Germany., Schulze A; Rentschler Biotechnologie GmbH, Erwin-Rentschler-Straße 21, 88471, Laupheim, Germany., Krause F; Rentschler Biotechnologie GmbH, Erwin-Rentschler-Straße 21, 88471, Laupheim, Germany., Müller D; Rentschler Biotechnologie GmbH, Erwin-Rentschler-Straße 21, 88471, Laupheim, Germany.
Jazyk: angličtina
Zdroj: Biotechnology letters [Biotechnol Lett] 2016 Dec; Vol. 38 (12), pp. 2043-2049. Date of Electronic Publication: 2016 Sep 09.
DOI: 10.1007/s10529-016-2197-x
Abstrakt: 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.
Databáze: MEDLINE
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