title: High-level production of Fc-fused kringle domain in Pichia pastoris.
abstract: Recently, as a new non-immunoglobulin-based protein scaffold, a human kringle domain was successfully engineered toward biologically functional agonists and antagonists. In this study, the fed-batch cultivation conditions were optimized for enhanced production of an Fc-fused kringle domain (KD548-Fc) in Pichia pastoris. Fed-batch cultivations were performed in 5-l laboratory-scale bioreactors, and in order to find the optimal conditions for high-level production of KD548-Fc, several parameters including the initial carbon source (glycerol) concentration, temperature, and pH were investigated. When cells were cultivated at pH 4.0 and 25 °C with 9.5 % glycerol in the initial medium, the highest production yield (635 mg/l) was achieved with high productivity (7.2 mg/l/h). Furthermore, functional KD548-Fc was successfully purified from the culture broth using a simple purification procedure with high purity and recovery yield.
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, 335 Gwahagno, Yuseong-gu, Daejeon, 305-701, Republic of Korea.
authors.forename: Gu Min
authors.initials: GM
authors.lastname: Jeong
authors.forename: Yong Jae
authors.initials: YJ
authors.lastname: Lee
authors.forename: Yong Sung
authors.initials: YS
authors.lastname: Kim
authors.forename: Ki Jun
authors.initials: KJ
authors.lastname: Jeong
mesh_terms: D019149:Bioreactors; D003470:Culture Media; D005285:Fermentation; D005990:Glycerol / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007141:Immunoglobulin Fc Fragments / Q000235:genetics; D018082:Kringles / Q000235:genetics*; D010843:Pichia / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D013696:Temperature
chemical_list: D003470:Culture Media; D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins; D005990:Glycerol