title: Modified harvest system for enhancing Factor VIII yield in alternating tangential flow perfusion culture.
abstract: This study describes the development and experimental verification of a modified harvest system to enhance Factor VIII (FVIII) yield in an alternating tangential flow (ATF) perfusion culture. The main innovation of the modified harvest system is the use of check and pinch valves, eliminating the need of a peristaltic pump for harvest. The system was applied to perfusion cultures of Chinese hamster ovary cells, which co-express both recombinant human FVIII (rhFVIII) and von Willebrand factor (vWF). The modified harvest system showed comparable cell growth with the conventional harvest system using a peristaltic pump. The perfusion rate was successfully controlled using the system. In addition, the modified harvest system achieved an approximately 13.6-fold increase in the final concentration yield of FVIII activity and a 1.47-fold increase in the production yield of FVIII activity compared with a peristaltic pump. Enhancement of the yield of FVIII activity resulted from the reduction of FVIII antigen (
  Ag) retention. As a result of transmembrane pressure (TMP) measurement, the reduction of the retained
  Ag was due to the increased TMP, which was caused by the characteristic function of a check valve, compared with a peristaltic harvest system. The modified harvest system developed in this study could be useful to enhance the production yield of other recombinant proteins in ATF perfusion culture.
authors.affiliation: Research Institute, Dong-A Socio-Holdings Co., Ltd., Yong-in 449-900, Republic of Korea; Interdisciplinary Program for Bioengineering, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Seung-Chul
authors.initials: SC
authors.lastname: Kim
authors.affiliation: Research Institute, Dong-A Socio-Holdings Co., Ltd., Yong-in 449-900, Republic of Korea.
authors.forename: Sora
authors.initials: S
authors.lastname: An
authors.affiliation: Research Institute, Dong-A Socio-Holdings Co., Ltd., Yong-in 449-900, Republic of Korea.
authors.forename: Hyun-Ki
authors.initials: HK
authors.lastname: Kim
authors.affiliation: Research Institute, Dong-A Socio-Holdings Co., Ltd., Yong-in 449-900, Republic of Korea.
authors.forename: Beom-Soo
authors.initials: BS
authors.lastname: Park
authors.affiliation: Research Institute, Dong-A Socio-Holdings Co., Ltd., Yong-in 449-900, Republic of Korea.
authors.forename: Kyu-Heum
authors.initials: KH
authors.lastname: Na
authors.affiliation: Interdisciplinary Program for Bioengineering, Seoul National University, Seoul 151-742, Republic of Korea. Electronic address: byungkim@snu.ac.kr.
authors.forename: Byung-Gee
authors.initials: BG
authors.lastname: Kim
mesh_terms: D000818:Animals; D057219:Biofouling / Q000517:prevention & control; D001709:Biotechnology / Q000295:instrumentation* / Q000379:methods*; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D005169:Factor VIII / Q000096:biosynthesis*; D006801:Humans; D010477:Perfusion / Q000295:instrumentation / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014841:von Willebrand Factor / Q000096:biosynthesis
chemical_list: D011994:Recombinant Proteins; D014841:von Willebrand Factor; D005169:Factor VIII
keywords: Alternating tangential flow perfusion culture; Check valve; FVIII yield; Filter fouling; Modified harvest system; Pinch valve; Transmembrane pressure