High-Throughput Process Development for Recombinant Human Serum Albumin Separation from Pichia pastoris Broth with Mixed-Mode Chromatography

Abstract

Due to the shortage of blood donation and safe issues of blood-derived pathogens, recombinant human serum albumin (rHSA) expressed by Pichia pastoris is a potential substitute for plasma-derived HSA (pHSA). In this work, four commercialized mixed-mode resins (Capto adhere, Capto MMC, MX-Trp-650M, and Nuvia cPrime) and two ion-exchange resins (Q Sepharose FF and SP Sepharose FF) were used for rHSA separation from Pichia pastoris cell culture broth, and loading and elution conditions were optimized via a high-throughput process using microtiter filter plates. The results indicated that resins with cation-exchange groups had better adsorption performance. The loading conditions at pH 4.0 and dilution rate of 1.0 were suitable for SP Sepharose FF, and no dilution worked for mixed-mode resins. The optimal elution pH was determined to be pH 7.0 for SP Sepharose FF, pH 8.0 for MX-Trp-650M and Nuvia cPrime, and pH 9.0 for Capto MMC. Moreover, rHSA separation from Pichia pastoris broth was performed on column unde...