PMID- 32642835
OWN - NLM
STAT- MEDLINE
DCOM- 20210423
LR  - 20210628
IS  - 1618-2650 (Electronic)
IS  - 1618-2642 (Linking)
VI  - 412
IP  - 23
DP  - 2020 Sep
TI  - Identification and semi-relative quantification of intact glycoforms by 
      nano-LC-(Orbitrap)MS: application to the alpha-subunit of human chorionic 
      gonadotropin and follicle-stimulating hormone.
PG  - 5729-5741
LID - 10.1007/s00216-020-02794-3 [doi]
AB  - Human chorionic gonadotropin (hCG) and follicle-stimulating hormone (FSH) belong 
      to the family of glycoprotein polypeptide hormones called gonadotropins. They are 
      heterodimers sharing the alpha-subunit structure that has 2 N-glycosylation sites. A 
      method based on nano-reversed-phase liquid chromatography coupled to 
      high-resolution mass spectrometry with an Orbitrap analyzer was developed for the 
      first time to characterize the glycosylation state of the alpha-subunit at the intact 
      level. A recombinant hCG-based drug, Ovitrelle(R), was analyzed. This method 
      combined with an appropriate data treatment allowed the detection of not only the 
      major isoforms but also the minority ones with a high mass accuracy. More than 30 
      hCGalpha glycoforms were detected without overlapping of the isotopic patterns. The 
      figures of merit of the method were assessed. The relative standard deviations 
      (RSDs) of the retention time ranged between 0.1 and 6.08% (n = 3), with an 
      average of 0.4%. The RSDs of the peak area measured on the extracted ion 
      chromatogram of each glycoform are below 38% (n = 3), with an average of 16%, 
      thus allowing semi-relative quantification. The ability to accurately profile 
      glycosylated variants of hCGalpha was next demonstrated by comparing qualitatively 
      and semi-quantitatively 3 batches of Ovitrelle(R). The method was also used to 
      analyze 3 batches of a recombinant FSH-based drug, Puregon(R), and 30 FSHalpha 
      glycoforms were detected and semi-quantified. This demonstrates the high 
      potential of this method for fast quality control or comparison of the 
      glycosylation of glycoprotein-based pharmaceutical preparations. Graphical 
      abstract.
FAU - Al Matari, Amira
AU  - Al Matari A
AD  - Laboratory of Analytical, Bioanalytical Sciences and Miniaturization, Chemistry, 
      Biology and Innovation (CBI) UMR 8231, ESPCI Paris PSL, CNRS, PSL Research 
      University, 10 rue Vauquelin, 75005, Paris, France.
FAU - Combes, Audrey
AU  - Combes A
AD  - Laboratory of Analytical, Bioanalytical Sciences and Miniaturization, Chemistry, 
      Biology and Innovation (CBI) UMR 8231, ESPCI Paris PSL, CNRS, PSL Research 
      University, 10 rue Vauquelin, 75005, Paris, France.
FAU - Camperi, Julien
AU  - Camperi J
AD  - Laboratory of Analytical, Bioanalytical Sciences and Miniaturization, Chemistry, 
      Biology and Innovation (CBI) UMR 8231, ESPCI Paris PSL, CNRS, PSL Research 
      University, 10 rue Vauquelin, 75005, Paris, France.
FAU - Fournier, Thierry
AU  - Fournier T
AD  - Universite de Paris, INSERM, <<Pathophysiology & Pharmacotoxicology of the Human 
      Placenta, pre & postnatal Microbiota>>, 3PHM, 75006, Paris, France.
FAU - Pichon, Valerie
AU  - Pichon V
AD  - Laboratory of Analytical, Bioanalytical Sciences and Miniaturization, Chemistry, 
      Biology and Innovation (CBI) UMR 8231, ESPCI Paris PSL, CNRS, PSL Research 
      University, 10 rue Vauquelin, 75005, Paris, France.
AD  - Sorbonne Universite, 4 place Jussieu, 75005, Paris, France.
FAU - Delaunay, Nathalie
AU  - Delaunay N
AD  - Laboratory of Analytical, Bioanalytical Sciences and Miniaturization, Chemistry, 
      Biology and Innovation (CBI) UMR 8231, ESPCI Paris PSL, CNRS, PSL Research 
      University, 10 rue Vauquelin, 75005, Paris, France. nathalie.delaunay@espci.fr.
LA  - eng
GR  - ANR-10-LABX-31/Agence Nationale de la Recherche/
GR  - grant to Amira Al Matari/association Al Ihssan/
GR  - PME DIM Analytics/Conseil Regional Ile de France/
PT  - Journal Article
DEP - 20200708
PL  - Germany
TA  - Anal Bioanal Chem
JT  - Analytical and bioanalytical chemistry
JID - 101134327
RN  - 0 (Chorionic Gonadotropin)
RN  - 9002-68-0 (Follicle Stimulating Hormone)
SB  - IM
MH  - Animals
MH  - Chorionic Gonadotropin/*analysis
MH  - Chromatography, High Pressure Liquid/*methods
MH  - Cricetinae
MH  - Follicle Stimulating Hormone/*analysis
MH  - Glycosylation
MH  - Humans
MH  - Mass Spectrometry/*methods
MH  - Mice
OTO - NOTNLM
OT  - Follicle-stimulating hormone
OT  - Glycoprotein
OT  - Human chorionic gonadotropin
OT  - Intact protein
OT  - Mass spectrometry
OT  - Nano-liquid chromatography
EDAT- 2020/07/10 06:00
MHDA- 2021/04/24 06:00
CRDT- 2020/07/10 06:00
PHST- 2020/05/14 00:00 [received]
PHST- 2020/06/26 00:00 [accepted]
PHST- 2020/06/16 00:00 [revised]
PHST- 2020/07/10 06:00 [pubmed]
PHST- 2021/04/24 06:00 [medline]
PHST- 2020/07/10 06:00 [entrez]
AID - 10.1007/s00216-020-02794-3 [pii]
AID - 10.1007/s00216-020-02794-3 [doi]
PST - ppublish
SO  - Anal Bioanal Chem. 2020 Sep;412(23):5729-5741. doi: 10.1007/s00216-020-02794-3. 
      Epub 2020 Jul 8.