PMID- 20932526
OWN - NLM
STAT- MEDLINE
DCOM- 20110121
LR  - 20191210
IS  - 1873-3778 (Electronic)
IS  - 0021-9673 (Linking)
VI  - 1217
IP  - 45
DP  - 2010 Nov 5
TI  - Charge heterogeneity of a therapeutic monoclonal antibody conjugated with a 
      cytotoxic antitumor antibiotic, calicheamicin.
PG  - 7164-71
LID - 10.1016/j.chroma.2010.09.022 [doi]
AB  - A robust and highly reproducible capillary isoelectric focusing (cIEF) method for 
      the evaluation of charge heterogeneity of monoclonal antibody (mAb) 
      pharmaceutical which contains covalently bound antitumor compounds was developed 
      using a combination of commercially available dimethylpolysiloxane-coated 
      capillary and carrier ampholyte. In order to optimize major analytical parameters 
      for robust mobilization, experimental responses from three pI markers were 
      selected. The optimized method gave excellent repeatability and intermediate 
      precision in estimated pI values of charge variants with relative standard 
      deviations (RSDs) of not more than 0.06% and 0.95%, respectively, when using 
      IgG(4) as a model. Furthermore, RSDs of charge variant compositions were less 
      than 5.0%. These results suggest that the proposed method can be a powerful tool 
      for reproducible evaluation of charge variants of both naked mAbs and their 
      conjugates with high resolution, and it is applicable to quality testing and 
      detailed characterization in the pharmaceutical industry. In addition, it should 
      be noticed that the method provided non-linear pH gradient within the tested 
      ranges, from pI 9.50 to 3.78, and the pH gradient caused the inconsistency of 
      estimated pI ranges between cIEF and gel IEF. This result indicates that 
      selecting appropriate pI markers based on the target pI ranges of charge variants 
      for each mAb related pharmaceutical is highly recommended for the precise 
      determination of pI values.
CI  - Copyright (c) 2010 Elsevier B.V. All rights reserved.
FAU - Maeda, Eiki
AU  - Maeda E
AD  - Analytical Development Laboratories, CMC Center, Takeda Pharmaceutical Company 
      Limited, Juso-honmachi 2-17-85, Yodogawa-ku, Osaka 532-8686, Japan. maeda 
      eiki@takeda.co.jp
FAU - Urakami, Koji
AU  - Urakami K
FAU - Shimura, Kiyohito
AU  - Shimura K
FAU - Kinoshita, Mitsuhiro
AU  - Kinoshita M
FAU - Kakehi, Kazuaki
AU  - Kakehi K
LA  - eng
PT  - Journal Article
DEP - 20100917
PL  - Netherlands
TA  - J Chromatogr A
JT  - Journal of chromatography. A
JID - 9318488
RN  - 0 (Aminoglycosides)
RN  - 0 (Antibiotics, Antineoplastic)
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antibodies, Monoclonal, Humanized)
RN  - 0 (Immunotoxins)
RN  - 93NS566KF7 (Gemtuzumab)
SB  - IM
MH  - Aminoglycosides/*chemistry/metabolism
MH  - Antibiotics, Antineoplastic/*chemistry/metabolism
MH  - Antibodies, Monoclonal/*chemistry/metabolism
MH  - Antibodies, Monoclonal, Humanized
MH  - Electrophoresis, Capillary/*methods
MH  - Gemtuzumab
MH  - Glycosylation
MH  - Immunoelectrophoresis/*methods
MH  - Immunotoxins/*chemistry/metabolism
MH  - Isoelectric Point
MH  - Models, Chemical
MH  - Regression Analysis
MH  - Reproducibility of Results
EDAT- 2010/10/12 06:00
MHDA- 2011/01/22 06:00
CRDT- 2010/10/12 06:00
PHST- 2010/07/08 00:00 [received]
PHST- 2010/09/01 00:00 [revised]
PHST- 2010/09/08 00:00 [accepted]
PHST- 2010/10/12 06:00 [entrez]
PHST- 2010/10/12 06:00 [pubmed]
PHST- 2011/01/22 06:00 [medline]
AID - S0021-9673(10)01242-2 [pii]
AID - 10.1016/j.chroma.2010.09.022 [doi]
PST - ppublish
SO  - J Chromatogr A. 2010 Nov 5;1217(45):7164-71. doi: 10.1016/j.chroma.2010.09.022. 
      Epub 2010 Sep 17.