PMID- 20730829
OWN - NLM
STAT- MEDLINE
DCOM- 20110113
LR  - 20191210
IS  - 1615-9314 (Electronic)
IS  - 1615-9306 (Linking)
VI  - 33
IP  - 17-18
DP  - 2010 Sep
TI  - Rapid and refined separation of human IgG2 disulfide isomers using superficially 
      porous particles.
PG  - 2671-80
LID - 10.1002/jssc.201000230 [doi]
AB  - A rapid reversed-phase HPLC separation of recombinant human immunoglobulin gamma 
      2 (IgG2) disulfide isomers using columns packed with superficially porous 
      particles is reported. Under optimal conditions, a separation of monoclonal IgG2 
      disulfide isomers was achieved in 10 min using a Poroshell 300SB-C8 column via a 
      combination of high column temperature (85 degrees C), mobile phases with high eluotropic 
      strength (e.g. isopropanol) and high flow rate (1.5 mL/min). Thermodynamic 
      stability analyses of chromatographically enriched IgG2 disulfide isomers 
      revealed differences in their individual denaturation temperatures, which 
      correlate with the observed temperature-dependent refinement of peak profiles by 
      reversed-phase HPLC. This reversed-phase HPLC method in conjunction with other 
      orthogonal analytical techniques (e.g. capillary gel electrophoresis, peptide 
      mapping, ion exchange chromatography, etc.) is being used to characterize 
      disulfide isomers in the development of therapeutic IgG2 antibodies.
FAU - Wang, Qian
AU  - Wang Q
AD  - Analytical R&D, Global Biologics, Pfizer Inc., Chesterfield, MO 63017, USA. 
      qian.wang2@pfizer.com
FAU - Lacher, Nathan A
AU  - Lacher NA
FAU - Muralidhara, Bilikallahalli K
AU  - Muralidhara BK
FAU - Schlittler, Michael R
AU  - Schlittler MR
FAU - Aykent, Serdar
AU  - Aykent S
FAU - Demarest, Charles W
AU  - Demarest CW
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PL  - Germany
TA  - J Sep Sci
JT  - Journal of separation science
JID - 101088554
RN  - 0 (Disulfides)
RN  - 0 (Immunoglobulin G)
SB  - IM
MH  - *Chromatography, High Pressure Liquid/instrumentation/methods
MH  - Disulfides/*chemistry
MH  - Humans
MH  - Immunoglobulin G/*chemistry/*isolation & purification
MH  - Isomerism
MH  - Models, Molecular
MH  - Porosity
MH  - Protein Conformation
MH  - Temperature
MH  - Thermodynamics
EDAT- 2010/08/24 06:00
MHDA- 2011/01/14 06:00
CRDT- 2010/08/24 06:00
PHST- 2010/08/24 06:00 [entrez]
PHST- 2010/08/24 06:00 [pubmed]
PHST- 2011/01/14 06:00 [medline]
AID - 10.1002/jssc.201000230 [doi]
PST - ppublish
SO  - J Sep Sci. 2010 Sep;33(17-18):2671-80. doi: 10.1002/jssc.201000230.