The Potential of Hydrocyclone Application for Mammalian Cell Separation in Perfusion Cultivation Bioreactors

Abstract

Hydrocyclones have been traditionally applied for long times in many industrial fields, such as in mineral processing and mining, chemical and petrochemical, and food industries. They have many characteristics that favor them as separation system in solid/liquid, gas/liquid and liquid/liquid processes. During the last two decades, they have been evaluated for their possible application in the separation of microbial and mammalian cells. Nowadays, mammalian cells are widely used for the production of a large number of valuable therapeutic proteins, antibodies, hormones and vaccines. This review highlights the potential of the application of hydrocyclones for mammalian cell separation in continuous perfusion biorecators. The discussion will cover the structure of hydrocyclone, mechanism of separation inside hydrocyclones, different theories describing the separation process, as well as the effect of changing different geometrical variables on the efficiency and performance of the separation process. Furthermore, we will focus on the latest developments achieved in the field of separation of living cells in both laboratory and pilot plant cultivation scales.

Hydrocyclones as cell retention device for CHO perfusion processes in single‐use bioreactors

“…In perfusion run #3 a customized bioreactor bag with large‐diameter ports allowed having an adequate diameter in the recirculation loop, that is, in the bioreactor bag connection to the HC feed and returning from the HC underflow orifice. This run was carried out for 20 days and reached a steady‐state at approximately 50 × 10 6 cells/ml, which is five‐fold higher than the maximum viable cell density that had been published so far for a perfusion process based on a HC as cell retention device (Elsayed, 2005). Total separation efficiencies of 95% and centrifugal separation efficiencies of 70–80% were obtained throughout the run.…”

“…A CHO cell suspension at 24 × 10 6 cells/ml was fed at 2.3 bar, resulting in a diluted overflow stream at 4.4 × 10 6 cells/ml, corresponding to a total separation efficiency (E T ) of 96% and a centrifugal efficiency (E′) of 82%. To our knowledge, this was the first time that a HC was challenged with a feed suspension at a viable cell density higher than 9.8 × 10 6 cells/ml (Elsayed, 2005), and the connector configuration proposed herein enabled a separation performance comparable to low concentration feed suspensions.…”

Optimising small hydrocyclone design using 3D printing and CFD simulations

“…The application of small hydrocyclones for dewatering fine particles in bioprocesses is of growing interest [1][2][3][4]. Since the diameter of the hydrocyclone determines largely the particle cut-size achieved [5], small diameter hydrocyclones can be used for particle separation in the micron range.…”