Progress in the Detection of Erythropoietin in Blood, Urine, and Tissue.
HIF2α
PHD inhibitor
Western blotting
deglycosylation
doping
erythropoietin
glycoprotein
hypoxia
Journal
Molecules (Basel, Switzerland)
ISSN: 1420-3049
Titre abrégé: Molecules
Pays: Switzerland
ID NLM: 100964009
Informations de publication
Date de publication:
30 May 2023
30 May 2023
Historique:
received:
30
03
2023
revised:
22
05
2023
accepted:
24
05
2023
medline:
12
6
2023
pubmed:
10
6
2023
entrez:
10
6
2023
Statut:
epublish
Résumé
Detection of erythropoietin (Epo) was difficult until a method was developed by the World Anti-Doping Agency (WADA). WADA recommended the Western blot technique using isoelectric focusing (IEF)-PAGE to show that natural Epo and injected erythropoiesis-stimulating agents (ESAs) appear in different pH areas. Next, they used sodium N-lauroylsarcosinate (SAR)-PAGE for better differentiation of pegylated proteins, such as epoetin β pegol. Although WADA has recommended the use of pre-purification of samples, we developed a simple Western blotting method without pre-purification of samples. Instead of pre-purification, we used deglycosylation of samples before SDS-PAGE. The double detection of glycosylated and deglycosylated Epo bands increases the reliability of the detection of Epo protein. All of the endogenous Epo and exogenous ESAs shift to 22 kDa, except for Peg-bound epoetin β pegol. All endogenous Epo and exogenous ESAs were detected as 22 kDa deglycosylated Epo by liquid chromatography/mass spectrum (LC/MS) analysis. The most important factor for the detection of Epo is the selection of the antibody against Epo. WADA recommended clone AE7A5, and we used sc-9620. Both antibodies are useful for the detection of Epo protein by Western blotting.
Identifiants
pubmed: 37298922
pii: molecules28114446
doi: 10.3390/molecules28114446
pmc: PMC10254663
pii:
doi:
Substances chimiques
Erythropoietin
11096-26-7
Antibodies
0
Recombinant Proteins
0
Types de publication
Journal Article
Review
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : The Science Research Promotion Fund from Kitasato University Medical Center
ID : H24-01
Organisme : a Grant-in-Aid for Scientific Research from the Ministry of Education, Vulture, Sports, Science and Technology of Japan
ID : 18K08247
Références
Kidney Int. 2022 Aug;102(2):280-292
pubmed: 35644281
Ren Fail. 2021 Dec;43(1):803-810
pubmed: 33966598
J Sep Sci. 2017 Dec;40(23):4636-4644
pubmed: 28988419
J Am Soc Nephrol. 2016 Aug;27(8):2265-9
pubmed: 26757994
Electrophoresis. 2020 Dec;41(23):2055-2061
pubmed: 32841408
Drug Test Anal. 2021 Dec 14;:
pubmed: 34905300
J Chromatogr A. 2021 Apr 26;1643:462043
pubmed: 33780879
J Biotechnol. 2014 Apr 20;176:29-39
pubmed: 24553072
J Clin Invest. 1992 Mar;89(3):753-60
pubmed: 1541670
Biochem Biophys Res Commun. 2014 Jun 27;449(2):222-8
pubmed: 24832733
Vitam Horm. 2017;105:57-77
pubmed: 28629525
Blood. 2008 May 15;111(10):5223-32
pubmed: 18202227
Lancet. 2016 Feb 27;387(10021):907-16
pubmed: 26314490
N Engl J Med. 2021 Apr 29;384(17):1589-1600
pubmed: 33913637
Blood. 1972 Nov;40(5):671-7
pubmed: 4637502
Anal Bioanal Chem. 2014 Feb;406(5):1317-29
pubmed: 23443522
N Engl J Med. 2021 Dec 16;385(25):2325-2335
pubmed: 34739194
J Pharmacol Exp Ther. 2020 Aug;374(2):342-353
pubmed: 32487538
J Cell Physiol. 2022 May;237(5):2420-2433
pubmed: 35014036
Eur J Biochem. 1990 Mar 10;188(2):405-11
pubmed: 2156701
Blood. 2011 Sep 1;118(9):2395-404
pubmed: 21652677
J Mol Histol. 2022 Apr;53(2):395-411
pubmed: 35084636
J Biol Chem. 1977 Aug 10;252(15):5558-64
pubmed: 18467
Physiol Rep. 2017 Dec;5(24):
pubmed: 29263117
Drug Test Anal. 2015 Nov-Dec;7(11-12):1014-6
pubmed: 26607413
Blood. 1989 Aug 1;74(2):622-8
pubmed: 2473803
Sci Rep. 2019 Aug 2;9(1):11254
pubmed: 31375751
Am J Kidney Dis. 2017 Jun;69(6):815-826
pubmed: 28242135
Int J Biol Macromol. 2007 Aug 1;41(3):354-7
pubmed: 17540444
Perit Dial Int. 2001 Jan-Feb;21(1):14-24
pubmed: 11280492
Am J Physiol. 1990 May;258(5 Pt 2):F1432-7
pubmed: 2337157
Anal Chem. 2007 Jun 15;79(12):4627-35
pubmed: 17500535
FASEB J. 2005 Dec;19(14):2023-5
pubmed: 16223784
Drug Test Anal. 2019 Sep;11(9):1290-1301
pubmed: 31232530
Eur J Clin Invest. 2005 Dec;35 Suppl 3:75-84
pubmed: 16281962
Cells. 2021 Feb 17;10(2):
pubmed: 33671138
Biochem Biophys Res Commun. 2008 Mar 21;367(4):834-9
pubmed: 18201556
Drug Dev Res. 2021 Sep;82(6):852-860
pubmed: 33480036
PLoS One. 2015 Jun 24;10(6):e0129484
pubmed: 26107632
J Am Soc Nephrol. 2002 Jul;13(7):1721-32
pubmed: 12089367
Adv Chronic Kidney Dis. 2019 Jul;26(4):253-266
pubmed: 31477256
N Engl J Med. 1974 Nov 14;291(20):1046-51
pubmed: 4606387
Nat Rev Nephrol. 2015 Jul;11(7):394-410
pubmed: 26055355
J Am Soc Nephrol. 2012 Oct;23(10):1631-4
pubmed: 22935483
Drug Test Anal. 2009 Nov;1(11-12):494-504
pubmed: 20355164
Drug Test Anal. 2022 Jan;14(1):181-187
pubmed: 33269539
Mol Pharmacol. 2014 Jun;85(6):898-908
pubmed: 24695083
Eur J Pharmacol. 2018 Nov 5;838:138-144
pubmed: 30179610
J Physiol. 2011 Mar 15;589(Pt 6):1251-8
pubmed: 21078592
Blood. 2000 Jan 1;95(1):111-9
pubmed: 10607693
Int J Mol Sci. 2023 Feb 03;24(3):
pubmed: 36769359
PLoS One. 2015 Apr 27;10(4):e0125215
pubmed: 25915923
Genes (Basel). 2021 Aug 16;12(8):
pubmed: 34440425
Pflugers Arch. 2016 Jan;468(1):3-12
pubmed: 26452589
Anal Bioanal Chem. 2011 Aug;401(2):463-81
pubmed: 21637931
Am J Physiol. 1992 Sep;263(3 Pt 2):F474-81
pubmed: 1415576
Am J Nephrol. 2008;28(4):628-40
pubmed: 18285684
Kidney Int. 1997 Feb;51(2):553-5
pubmed: 9027737
Eur J Pharmacol. 2021 Dec 5;912:174583
pubmed: 34678238
J Pharm Biomed Anal. 2016 Mar 20;121:181-187
pubmed: 26808067
J Biol Chem. 1991 Oct 25;266(30):20434-9
pubmed: 1657925
Ann Clin Lab Sci. 1981 Nov-Dec;11(6):484-7
pubmed: 7034633
Proc Natl Acad Sci U S A. 1983 Jun;80(12):3651-5
pubmed: 6190179
N Engl J Med. 2013 Jan 24;368(4):307-19
pubmed: 23343061
PLoS Comput Biol. 2023 Jan 24;19(1):e1010850
pubmed: 36693034
Nature. 1985 Feb 28-Mar 6;313(6005):806-10
pubmed: 3838366
Bioanalysis. 2012 Jul;4(13):1565-75
pubmed: 22831473
Am J Physiol Renal Physiol. 2010 Jul;299(1):F1-13
pubmed: 20444740
Blood Rev. 2013 Jan;27(1):41-53
pubmed: 23291219
Biochem Biophys Res Commun. 2018 Sep 18;503(4):3121-3127
pubmed: 30146260
Kidney Int. 2020 Apr;97(4):687-701
pubmed: 32033782
Clin Drug Investig. 2016 Sep;36(9):743-751
pubmed: 27352308
Adv Ther. 2019 Jun;36(6):1438-1454
pubmed: 30953333
J Clin Invest. 2000 Dec;106(11):1391-8
pubmed: 11104792
Front Horm Res. 2016;47:115-27
pubmed: 27348128
FEMS Microbiol Rev. 1994 Jul;14(3):253-8
pubmed: 7522023
J Pharm Biomed Anal. 2021 Feb 5;194:113750
pubmed: 33234415
N Engl J Med. 2013 Jan 24;368(4):320-32
pubmed: 23343062
Clin Exp Pharmacol Physiol. 2022 Feb;49(2):311-318
pubmed: 34653291
BMC Ophthalmol. 2019 Nov 14;19(1):227
pubmed: 31727007
Drug Test Anal. 2012 Nov;4(11):813-7
pubmed: 22566398
Biochem Biophys Res Commun. 2018 Jan 1;495(1):1317-1321
pubmed: 29191652
Molecules. 2022 Feb 08;27(3):
pubmed: 35164384
Nephrol Dial Transplant. 2015 Oct;30(10):1665-73
pubmed: 26238121
N Engl J Med. 2015 May 7;372(19):1832-43
pubmed: 25946282
J Cell Biochem. 2004 Sep 1;93(1):153-61
pubmed: 15352172
J Biol Chem. 1985 Nov 25;260(27):14727-31
pubmed: 4055799
Acta Physiol (Oxf). 2022 Aug;235(4):e13826
pubmed: 35491502
Methods Mol Biol. 2012;869:65-79
pubmed: 22585478
J Biol Chem. 1990 Jul 25;265(21):12127-30
pubmed: 2373681
Nature. 2000 Jun 8;405(6787):635
pubmed: 10864311
Heliyon. 2020 Nov 03;6(11):e05389
pubmed: 33195841
Molecules. 2021 Sep 05;26(17):
pubmed: 34500833
J Nephrol. 2022 Dec;35(9):2269-2282
pubmed: 36383211
Electrophoresis. 2007 Jun;28(12):1875-81
pubmed: 17503402
Arch Biochem Biophys. 2023 Jan 15;734:109487
pubmed: 36513130
Physiol Rep. 2020 Jun;8(12):e14485
pubmed: 32592328
Blood. 1999 Sep 15;94(6):1864-77
pubmed: 10477715
J Pharmacol Exp Ther. 2019 Dec;371(3):675-683
pubmed: 31585986
J Biol Chem. 1992 Apr 15;267(11):7703-9
pubmed: 1560005
J Am Soc Nephrol. 2015 Nov;26(11):2669-77
pubmed: 25817355
Nephron Clin Pract. 2008;110(2):c115-25
pubmed: 18931515
Infect Dis Clin North Am. 2001 Sep;15(3):877-99
pubmed: 11570146
Drug Test Anal. 2018 Nov;10(11-12):1698-1707
pubmed: 30365250