PMID- 20853347
OWN - NLM
STAT- MEDLINE
DCOM- 20110524
LR  - 20181201
IS  - 1520-6033 (Electronic)
IS  - 1520-6033 (Linking)
VI  - 26
IP  - 6
DP  - 2010 Nov-Dec
TI  - Downstream antibody purification using aqueous two-phase extraction.
PG  - 1662-70
LID - 10.1002/btpr.477 [doi]
AB  - The extraction of antibodies using a polyethylene glycol (PEG)-citrate aqueous 
      two-phase system (ATPS) was investigated. Studies using purified monoclonal 
      antibody (mAb) identified operating ranges for successful phase formation and 
      factors that significantly affected antibody partitioning. The separation of 
      antibody and host cell protein (HCP) from clarified cell culture media was 
      examined using statistical design of experiments (DOE). The partitioning of 
      antibody was nearly complete over the entire range of the operating space 
      examined. A model of the HCP partitioning was generated in which both NaCl and 
      citrate concentrations were identified as significant factors. To achieve the 
      highest purity, the partitioning of HCP from cell culture fluid into the product 
      containing phase was minimized using a Steepest Descent algorithm. An optimal 
      ATPS consisting of 14.0% (w/w) PEG, 8.4% (w/w) citrate, and 7.2% (w/w) NaCl at pH 
      7.2 resulted in a product yield of 89%, an approximate 7.6-fold reduction in HCP 
      levels relative to the clarified cell culture fluid before extraction and an 
      overall purity of 70%. A system consisting of 15% (w/w) PEG, 8% (w/w) citrate, 
      and 15% (w/w) NaCl at pH 5.5 reduced product-related impurities (aggregates and 
      low molecular product fragments) from  approximately 40% to less than 0.5% while achieving 95% 
      product recovery. At the experimental conditions that were optimized in the batch 
      mode, a scale-up model for the use of counter-current extraction technology was 
      developed to identify potential improvements in purity and recovery that could be 
      realized in the continuous operational mode.
CI  - Copyright (c) 2010 American Institute of Chemical Engineers (AIChE).
FAU - Mao, Lisong Nathan
AU  - Mao LN
AD  - Purification Process Development, Biogen IDEC Corporation, 5200 Research Place, 
      San Diego, CA 92122, USA. nathan.mao@biogenidec.com
FAU - Rogers, Jameson K
AU  - Rogers JK
FAU - Westoby, Matthew
AU  - Westoby M
FAU - Conley, Lynn
AU  - Conley L
FAU - Pieracci, John
AU  - Pieracci J
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Biotechnol Prog
JT  - Biotechnology progress
JID - 8506292
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Citrates)
RN  - 059QF0KO0R (Water)
RN  - 1Q73Q2JULR (Sodium Citrate)
RN  - 3WJQ0SDW1A (Polyethylene Glycols)
SB  - IM
MH  - Animals
MH  - Antibodies, Monoclonal/*isolation & purification
MH  - CHO Cells
MH  - Chemical Fractionation/*methods
MH  - Citrates/chemistry
MH  - Cricetinae
MH  - Cricetulus
MH  - Polyethylene Glycols/chemistry
MH  - Sodium Citrate
MH  - Water/chemistry
EDAT- 2010/09/21 06:00
MHDA- 2011/05/25 06:00
CRDT- 2010/09/21 06:00
PHST- 2010/09/21 06:00 [entrez]
PHST- 2010/09/21 06:00 [pubmed]
PHST- 2011/05/25 06:00 [medline]
AID - 10.1002/btpr.477 [doi]
PST - ppublish
SO  - Biotechnol Prog. 2010 Nov-Dec;26(6):1662-70. doi: 10.1002/btpr.477.