Control of specific growth rate for the enhanced production of human interferon α2b in glycoengineered Pichia pastoris: process analytical technology guided approach. Issue 10 (22nd July 2019)

• Record Type: Journal Article
• Authors: Katla, Srikanth; Mohan, Naresh; Pavan, Satya S; Pal, Uttariya; Sivaprakasam, Senthilkumar
• Is Part Of: Journal of chemical technology & biotechnology. Volume 94:Issue 10(2019)
• Page Start: 3111
• Page End: 3123
• Publication Date: 2019-07-22
• Subjects: process analytical technology -- capacitance -- glycoengineered Pichia pastoris -- human interferon alpha 2b -- specific growth rate estimator -- PID controller -- antiproliferative; Biotechnology -- Periodicals; Chemistry, Technical -- Periodicals; Chemical engineering -- Periodicals; Industries -- Environmental aspects -- Periodicals; 660
• DOI: 10.1002/jctb.6118
• Languages: English
• ISSNs: 0268-2575

Abstract

Abstract: BACKGROUND: Process variability in bioprocess systems involving genetically engineered strains is a common bottleneck and a real‐time insight of the on‐going process is crucial to achieve the desired product and its quality. In this study, a process analytical technology (PAT) platform was developed for the monitoring and control of specific growth rate during methanol induction phase ( μ met ) of glycoengineered Pichia pastoris fermentation for human interferon alpha 2b (huIFNα2b) production. RESULTS: The PAT guided approach involves real‐time monitoring of capacitance (Δ C ) facilitating online estimation of specific growth rate ( μ est ), which serves as a process input during controller application. Fed‐batch experiments using pulsed‐feeding of methanol at different dosage (20 g and 30 g) did not significantly influence μ met . Exponential methanol feeding was achieved using a modified proportional, integral and derivative (PID) controller for different predefined specific growth rate set point ( μ sp ) values, namely 0.015, 0.03, 0.04 and 0.06 h −1 . Exponential feeding strategy during the induction phase resulted in two crucial outcomes: (i) controlled methanol feeding rate (regulated by the developed PID controller) balanced the methanol consumption rate ( q s, met ) of the P. pastoris ; (ii) significant improvement in huIFNα2b titer (1483 mg L −1 ) and specific productivity(>0.4 mg g −1 . h) was achieved by the robust control of μ met at optimal (0.04 h −1 ) value. The purified huIFNα2b was found to exhibit antiproliferative effect against human breast cancer cell lines. CONCLUSIONS: Efficient control of μ met at a very low narrow range was achieved with a long‐term controller stability (>10 h) and the highest titer reported to date for huIFNα2b (at optimal μ met ) in the yeast expression platform. © 2019 Society of Chemical Industry