Biocalorimetric monitoring of glycoengineered P. pastoris cultivation for the production of recombinant huIFNα2b: A quantitative study based on mixed feeding strategies. (10th February 2020)

• Record Type: Journal Article
• Authors: Katla, Srikanth; Pavan, Satya Sai; Mohan, Naresh; Sivaprakasam, Senthilkumar
• Is Part Of: Biotechnology progress. Volume 36:Number 3(2020)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2020-02-10
• Subjects: biocalorimetry -- interferon alpha 2b -- mixed feeding -- P. pastoris -- process analytical technology; Biotechnology -- Periodicals; Food industry and trade -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/btpr.2971
• Languages: English
• ISSNs: 8756-7938

Abstract

Abstract: Real‐time monitoring of glycoengineered Pichia pastoris by employing process analytical technology (PAT) tools is vital for gaining deeper insights into the therapeutic protein production process. The present study focuses on influence of mixed feed carbon substrates during the induction phases of glycoengineered P . pastoris cultivation, for recombinant human interferon α2b (huIFNα2b) production by employing calorimetric (biological heat rate, q B ) and respirometric (oxygen uptake rate and carbon dioxide evolution rate) measurements. Mixed feed stream of carbon substrates (methanol + glycerol, methanol + sorbitol) at a predetermined "C‐molar ratios" were added during the induction phases. Methanol‐ and sorbitol‐based mixed feeding approach resulted in an improved huIFNα2b titer of 288 mg/L by channeling of methanol predominantly towards an optimal functioning of AOX expression system. A stand‐off between biomass yield Y X S and biomass heat yield Y Q X coefficient, degree of reduction of methanol and its cosubstrate (glycerol and sorbitol) determines the fraction of carbon energy channeled toward biomass and protein production, under strict aerobic conditions. Calorespirometric monitoring and assessment of thermal yields enables a reliable prediction of process variables, leading to futuristic efficient PAT‐based feed rate control.