title: Integrated Microfluidic Lectin Barcode Platform for High-Performance Focused Glycomic Profiling.
abstract: Protein glycosylation is one of the key processes that play essential roles in biological functions and dysfunctions. However, progress in glycomics has considerably lagged behind genomics and proteomics, due in part to the enormous challenges in analysis of glycans. Here we present a new integrated and automated microfluidic lectin barcode platform to substantially improve the performance of lectin array for focused glycomic profiling. The chip design and flow control were optimized to promote the lectin-glycan binding kinetics and speed of lectin microarray. Moreover, we established an on-chip lectin assay which employs a very simple blocking method to effectively suppress the undesired background due to lectin binding of antibodies. Using this technology, we demonstrated focused differential profiling of tissue-specific glycosylation changes of a biomarker, CA125 protein purified from ovarian cancer cell line and different tissues from ovarian cancer patients in a fast, reproducible, and high-throughput fashion. Highly sensitive CA125 detection was also demonstrated with a detection limit much lower than the clinical cutoff value for cancer diagnosis. This microfluidic platform holds the potential to integrate with sample preparation functions to construct a fully integrated "sample-to-answer" microsystem for focused differential glycomic analysis. Thus, our technology should present a powerful tool in support of rapid advance in glycobiology and glyco-biomarker development.
authors.affiliation: Department of Chemistry, University of Kansas, Lawrence, KS 66045.
authors.forename: Yuqin
authors.initials: Y
authors.lastname: Shang
authors.affiliation: College of Water Resource and Hydropower, Sichuan Agricultural University, Ya'an, Sichuan 625014, P.R. China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Zeng
authors.affiliation: Department of Chemistry, University of Kansas, Lawrence, KS 66045.|University of Kansas Cancer Center, Kansas City, KS 66160.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zeng
mesh_terms: D015415:Biomarkers; D054794:Glycomics / Q000379:methods*; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D044085:Microfluidics / Q000379:methods*; D011134:Polysaccharides / Q000378:metabolism*; D040081:Protein Array Analysis / Q000379:methods; D011485:Protein Binding; D040901:Proteomics / Q000379:methods
chemical_list: D015415:Biomarkers; D006023:Glycoproteins; D037102:Lectins; D011134:Polysaccharides
references.citation: Biomark Med. 2007 Dec;1(4):513-23
references.pmid: 20477371
references.citation: Adv Carbohydr Chem Biochem. 2011;65:219-71
references.pmid: 21763513
references.citation: Hum Reprod. 2005 Sep;20(9):2632-8
references.pmid: 15905287
references.citation: Anal Bioanal Chem. 2009 May;394(2):469-79
references.pmid: 19280179
references.citation: J Biochem Biophys Methods. 2007 Apr 10;70(3):415-26
references.pmid: 17112594
references.citation: JALA Charlottesv Va. 2010 Dec 1;15(6):455-463
references.pmid: 21218162
references.citation: Lab Chip. 2005 Oct;5(10):1096-103
references.pmid: 16175266
references.citation: Proteomics. 2014 Mar;14(4-5):525-46
references.pmid: 24339177
references.citation: Anal Chem. 2008 Nov 15;80(22):8822-7
references.pmid: 18855407
references.citation: Anal Bioanal Chem. 2013 Jul;405(17):5743-58
references.pmid: 23568613
references.citation: J Clin Immunol. 2010 May;30 Suppl 1:S15-9
references.pmid: 20390325
references.citation: J Cancer Res Clin Oncol. 2008 Aug;134(8):851-60
references.pmid: 18264723
references.citation: J Biol Chem. 2003 Aug 1;278(31):28619-34
references.pmid: 12734200
references.citation: Anal Methods. 2011 Jan 1;3(1):20-32
references.pmid: 32938106
references.citation: Proc Natl Acad Sci U S A. 2013 Jan 8;110(2):477-82
references.pmid: 23267104
references.citation: Lab Chip. 2008 Dec;8(12):2062-70
references.pmid: 19023469
references.citation: Mol Cell Proteomics. 2009 Jan;8(1):99-108
references.pmid: 18697734
references.citation: Anal Chem. 2007 Sep 15;79(18):7170-5
references.pmid: 17685584
references.citation: Glycobiology. 2004 May;14(5):373-86
references.pmid: 14736727
references.citation: J Biol Chem. 1989 Feb 15;264(5):2415-23
references.pmid: 2536709
references.citation: J Proteome Res. 2013 Oct 4;12(10):4490-6
references.pmid: 23984816
references.citation: Lab Chip. 2013 Nov 7;13(21):4190-7
references.pmid: 23989677
references.citation: Anal Chem. 2013 Mar 19;85(6):3127-35
references.pmid: 23398125
references.citation: Nat Chem Biol. 2010 Oct;6(10):713-23
references.pmid: 20852609
references.citation: Proteomics. 2008 Aug;8(16):3210-20
references.pmid: 18690643
references.citation: Lab Chip. 2013 Apr 7;13(7):1413-21
references.pmid: 23407647
references.citation: Anal Chem. 2010 Apr 15;82(8):3183-90
references.pmid: 20192178
references.citation: Anal Chem. 2011 Mar 15;83(6):2279-85
references.pmid: 21344864
references.citation: Nat Methods. 2011 Jan;8(1):85-90
references.pmid: 21131969
references.citation: Cancer Biomark. 2008;4(1):35-42
references.pmid: 18334732
references.citation: Nat Methods. 2007 May;4(5):437-44
references.pmid: 17417647
references.citation: J Biochem. 2008 Aug;144(2):139-47
references.pmid: 18390573
references.citation: Nat Methods. 2005 Nov;2(11):851-6
references.pmid: 16278656
references.citation: Anal Biochem. 2006 Jan 15;348(2):284-93
references.pmid: 16307717
references.citation: Talanta. 2011 Jun 15;84(5):1284-90
references.pmid: 21641439
references.citation: Nat Biotechnol. 2006 Sep;24(9):1100-1
references.pmid: 16964219
references.citation: Biomed Microdevices. 2007 Aug;9(4):545-54
references.pmid: 17505888
references.citation: Nat Biotechnol. 2008 Apr;26(4):417-26
references.pmid: 18392027
references.citation: Int J Mol Sci. 2013 Jul 26;14(8):15636-54
references.pmid: 23896595
references.citation: Anal Chem. 2012 Feb 7;84(3):1425-31
references.pmid: 22196688
references.citation: Glycobiology. 2000 May;10(5):477-86
references.pmid: 10764836
references.citation: Anal Chem. 2009 Jul 1;81(13):5413-20
references.pmid: 19514701
references.citation: Nature. 2015 Jul 9;523(7559):177-82
references.pmid: 26106858
references.citation: Glycobiology. 2007 Dec;17(12):1344-56
references.pmid: 17884841
references.citation: Chem Soc Rev. 2013 May 21;42(10):4443-58
references.pmid: 23443201
references.citation: Anal Chim Acta. 2011 Sep 2;701(1):6-14
references.pmid: 21763802
references.citation: J Chromatogr A. 2004 Oct 22;1053(1-2):79-88
references.pmid: 15543974
references.citation: Biosens Bioelectron. 2007 Feb 15;22(7):1403-9
references.pmid: 16879959
references.citation: Proteomics. 2009 Feb;9(4):882-913
references.pmid: 19212958
references.citation: J Proteome Res. 2013 Mar 1;12(3):1408-18
references.pmid: 23360124
references.citation: Annu Rev Anal Chem (Palo Alto Calif). 2015;8:463-83
references.pmid: 26070719
references.citation: Talanta. 2011 Apr 15;84(2):565-71
references.pmid: 21376988
references.citation: Biomicrofluidics. 2014 Jun 06;8(3):034107
references.pmid: 24959308
references.citation: Anal Chem. 2010 Jun 15;82(12):5095-106
references.pmid: 20491449