PMID- 26786498
OWN - NLM
STAT- MEDLINE
DCOM- 20171227
LR  - 20180213
IS  - 1460-2423 (Electronic)
IS  - 0959-6658 (Linking)
VI  - 26
IP  - 6
DP  - 2016 Jun
TI  - Profiling N-linked oligosaccharides from IgG by high-performance anion-exchange 
      chromatography with pulsed amperometric detection.
PG  - 582-91
LID - 10.1093/glycob/cww006 [doi]
AB  - Understanding and characterizing protein therapeutic glycosylation is important 
      with growing evidence that glycosylation impacts biological efficacy, 
      pharmacokinetics and cellular toxicity. Protein expression systems and reactor 
      conditions can impact glycosylation, leading to potentially undesirable 
      glycosylation. For example, high-mannose species may be present, which are 
      atypical of human antibody glycosylation. Their presence in the Fc domain has 
      been linked to increased serum clearance of immunoglobulin G (IgG) antibodies. 
      High-performance anion-exchange chromatography with pulsed amperometric detection 
      (HPAE-PAD) is an effective tool for determining glycans present in glycoprotein 
      therapeutics. We report an improved HPAE-PAD method for IgG oligosaccharide 
      separation. The neutral glycans are well resolved, including separation of 
      high-mannose species from typical human IgG glycans. Oligosaccharide 
      identification was performed by comparison to known standards in conjunction with 
      selective exoglycosidase digestion of both standards and released glycans. 
      Retention times (RTs) of known glycans were compared with the retention times of 
      maltose, maltotriose and maltotetraose standards to define a retention index 
      value for each glycan. These retention indices were used to aid identification of 
      glycans from an example monoclonal antibody sample of unknown glycosylation. 
      Method ruggedness was evaluated across duplicate systems, analysts and triplicate 
      column lots. Comparing two systems with different analysts and columns, retention 
      time precision relative standard deviations (RSDs) were between 0.63 and 4.0% 
      while retention indices precision RSDs ranged from 0.27 to 0.56%. The separation 
      is orthogonal to capillary electrophoresis-based separation of labeled IgG 
      oligosaccharides.
CI  - (c) The Author 2016. Published by Oxford University Press. All rights reserved. For 
      permissions, please e-mail: journals.permissions@oup.com.
FAU - Rohrer, Jeffrey S
AU  - Rohrer JS
AD  - Thermo Fisher Scientific, 1214 Oakmead Parkway, Sunnyvale, CA 94085, USA 
      jeff.rohrer@thermofisher.com.
FAU - Basumallick, Lipika
AU  - Basumallick L
AD  - Thermo Fisher Scientific, 1214 Oakmead Parkway, Sunnyvale, CA 94085, USA.
FAU - Hurum, Deanna C
AU  - Hurum DC
AD  - Thermo Fisher Scientific, 1214 Oakmead Parkway, Sunnyvale, CA 94085, USA.
LA  - eng
PT  - Journal Article
DEP - 20160119
PL  - England
TA  - Glycobiology
JT  - Glycobiology
JID - 9104124
RN  - 0 (Immunoglobulin G)
RN  - 0 (Oligosaccharides)
RN  - EC 3.2.1.23 (beta-Galactosidase)
RN  - EC 3.2.1.51 (alpha-L-Fucosidase)
SB  - IM
MH  - Chromatography, High Pressure Liquid
MH  - Chromatography, Ion Exchange
MH  - Glycosylation
MH  - Humans
MH  - Hydrolysis
MH  - Immunoglobulin G/blood/*chemistry
MH  - Oligosaccharides/chemistry/*isolation & purification
MH  - Reproducibility of Results
MH  - alpha-L-Fucosidase/*chemistry
MH  - beta-Galactosidase/*chemistry
OTO - NOTNLM
OT  - HPAE-PAD
OT  - HPLC
OT  - N-linked glycans
OT  - glycoprotein
OT  - monoclonal antibodies
EDAT- 2016/01/21 06:00
MHDA- 2017/12/28 06:00
CRDT- 2016/01/21 06:00
PHST- 2015/09/10 00:00 [received]
PHST- 2016/01/08 00:00 [accepted]
PHST- 2016/01/21 06:00 [entrez]
PHST- 2016/01/21 06:00 [pubmed]
PHST- 2017/12/28 06:00 [medline]
AID - cww006 [pii]
AID - 10.1093/glycob/cww006 [doi]
PST - ppublish
SO  - Glycobiology. 2016 Jun;26(6):582-91. doi: 10.1093/glycob/cww006. Epub 2016 Jan 
      19.