PMID- 20408815
OWN - NLM
STAT- MEDLINE
DCOM- 20100824
LR  - 20161125
IS  - 1470-8744 (Electronic)
IS  - 0885-4513 (Linking)
VI  - 56
IP  - 1
DP  - 2010 May 19
TI  - Development of a rapid, high-efficiency, scalable refold for neurotrophin-4.
PG  - 27-34
LID - 10.1042/BA20090306 [doi]
AB  - A scalable refold for human neurotrophin-4 was developed as part of a 
      manufacturing process required for the production of supplies for preclinical and 
      clinical studies. The process redox system, chaotrope, solubilization additives, 
      pH, temperature and protein concentration were optimized. The limited 
      availability of suitable material for experimentation during concurrent 
      downstream process development led to the approach described in the present 
      paper: a combination of OFAT (one factor at a time) and multivariate DOE (design 
      of experiments) to identify appropriate conditions. The optimized refold 
      conditions included the use of sulfonated protein, raw materials utilized in 
      other process operations and an inexpensive redox system. The conditions were 
      found to be robust and were demonstrated from the millilitre scale to the 300 
      litre pilot scale. A process control procedure that utilized an RPC 
      (reversed-phase chromatography) quantitative assay to monitor the percentage 
      conversion into oxidized protein was developed. Refold conversions of 80-90% were 
      obtained under ambient temperature and atmospheric conditions, with reaction 
      times of approx. 18 h.
FAU - Buckley, John J
AU  - Buckley JJ
AD  - Pfizer, Inc., Chesterfield, MO 63017, U.S.A. john.joseph.buckley@pfizer.com
FAU - Hoeltzli, Sydney D
AU  - Hoeltzli SD
FAU - Johnson, Gary V
AU  - Johnson GV
LA  - eng
PT  - Journal Article
DEP - 20100519
PL  - United States
TA  - Biotechnol Appl Biochem
JT  - Biotechnology and applied biochemistry
JID - 8609465
RN  - 0 (Nerve Growth Factors)
RN  - P658DCA9XD (neurotrophin 4)
SB  - IM
MH  - Escherichia coli/genetics
MH  - Humans
MH  - Hydrogen-Ion Concentration
MH  - Industrial Microbiology/economics/*methods
MH  - Nerve Growth Factors/*biosynthesis/chemistry/isolation & purification
MH  - Oxidation-Reduction
MH  - Protein Folding
MH  - Solubility
MH  - Temperature
EDAT- 2010/04/23 06:00
MHDA- 2010/08/25 06:00
CRDT- 2010/04/23 06:00
PHST- 2010/04/23 06:00 [entrez]
PHST- 2010/04/23 06:00 [pubmed]
PHST- 2010/08/25 06:00 [medline]
AID - BA20090306 [pii]
AID - 10.1042/BA20090306 [doi]
PST - epublish
SO  - Biotechnol Appl Biochem. 2010 May 19;56(1):27-34. doi: 10.1042/BA20090306.