Title

A new single-use disk stack separator for pilot scale CHO cell-based antibody productions

Conference Dates

March 20 – 23, 2022

Abstract

Single-use systems are well-established in modern antibody productions. This is especially true for upstream processing (cell production, fermentation and cell harvesting), where complete single-use lines have become reality [1]. However, it has proven rather difficult to convert centrifugation for cell harvesting to single-use technologies, even though Sartorius and PneumaticScale Angelus have had solutions available for some time with their KSep (fluidized bed technology) and Carr Centritech UniFuge (conventional separation) systems. Previously, there were no single-use disk stack separators on the market. However, such a system, the kytero® 500, has been developed by GEA and was launched in summer 2021.

This poster summarizes the results of tests of the kytero® 500 with CHO suspension cells. The biomass to be separated was from fed-batch cultivations of immunoglobulin G (IgG) producing ExpiCHO-S cells in a 200 L BIOSTAT STR from Sartorius. The cells were cultivated with Gibco’s chemically defined media (Efficient-ProTM Medium and Efficient-ProTM Feed 2). Separation of the biomass from the culture broth was performed at a living cell density of 17 x 106 cells mL-1 and a viability of 98.9%. To study the performance of this new centrifuge over as wide a range as possible, various separation parameters, including separator speed, feed flow, and biomass concentration in concentrate, were tested.

Regardless of the separator settings, no solid particles were detected in the centrate, which was also confirmed by turbidity measurements. Nephelometric Turbidity Unit-based clarification efficiencies ranged from 96.6% to 97.5%. In addition, no increase in DNA or Host Cell Protein concentration in the IgG-containing centrate phase and no loss of IgG were detected. The biomass was concentrated to a maximum of 30-fold, yielding 99.3% of the liquid phase. The kytero® 500 is characterized by its ease of use and compact design. The highly efficient cell separation delivered by the kytero® 500 may result in a smaller depth filter area in product purification processes and thus a reduction in process costs.

1. BioPlan Associates, 17th annual report and survey of biopharmaceutical manufacturing capacity and production: A study of biotherapeutic developers and contract manufacturing organizations, 2020.

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