title: Maximizing recombinant human serum albumin production in a Mut(s) Pichia pastoris strain.
abstract: Human serum albumin (HSA) is a cysteine rich molecule that is most abundant in human blood plasma. To remain viable in the market due to lower marketing costs for HSA, it is important to produce a large quantity in an economical manner by recombinant technology. The objective of this study was to maximize recombinant HSA (rHSA) production using a Mut(s) Pichia pastoris strain by fermentation process optimization. We evaluated the impact of process parameters on the production of rHSA, including induction cell density (wet cell weight, g/L) and the control of specific growth rate at induction. In this study, we demonstrated that induction cell density is a critical factor for high level production of rHSA under controlled specific growth rate. We observed higher specific productivities at higher induction cell densities (285 g/L) and at lower specific growth rates (0.0022-0.0024/h) during methanol induction phase, and achieved the broth titer of rHSA up to 10 g/L. The temperature shift from 24 to 28(o) C was effective to control the specific growth rate at low level (≤0.0024/h) during methanol induction phase while maintaining high specific productivity [0.0908 mgrHSA /(gwcw h)].
authors.affiliation: GlycoFi, Biologics Discovery, Merck & Co., Inc., Lebanon, NH.
authors.forename: Muralidhar
authors.initials: M
authors.lastname: Mallem
authors.forename: Shannon
authors.initials: S
authors.lastname: Warburton
authors.forename: Fang
authors.initials: F
authors.lastname: Li
authors.forename: Ishaan
authors.initials: I
authors.lastname: Shandil
authors.forename: Adam
authors.initials: A
authors.lastname: Nylen
authors.forename: Sehoon
authors.initials: S
authors.lastname: Kim
authors.forename: Youwei
authors.initials: Y
authors.lastname: Jiang
authors.forename: Michael
authors.initials: M
authors.lastname: Meehl
authors.forename: Marc
authors.initials: M
authors.lastname: d'Anjou
authors.forename: Terrance A
authors.initials: TA
authors.lastname: Stadheim
authors.forename: Byung-Kwon
authors.initials: BK
authors.lastname: Choi
mesh_terms: D001709:Biotechnology / Q000379:methods*; D055786:Gene Knockout Techniques; D006801:Humans; D010641:Phenotype; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012709:Serum Albumin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D012709:Serum Albumin
keywords: Pichia pastoris muts; cell density; human serum albumin; specific growth rate