title: Probing antigen-antibody interaction using fluorescence coupled capillary electrophoresis.
abstract: In this report, the use of fluorescence detection coupled capillary electrophoresis (CE-FL) allowed us to fully characterize the antigen-antibody interaction. CE-FL allowed separation of unbound quantum dots (QDs) and ligand bound QDs and also revealed an ordered assembly of biomolecules on QDs. Further, we observed FRET from QDs donor to DyLight acceptor, which were covalently conjugated with human IgG and goat anti-human IgG, respectively. The immunocomplex was formed and the mutual affinity of the antigen and antibody brought QDs and DyLight close enough to allow FRET to occur. This novel CE-based technique can be easily extended to other FRET systems based on QDs and may have potential application in the detection of antibodies.
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou 213164, China. jiangxia@gmail.com.
authors.forename: Jianhao
authors.initials: J
authors.lastname: Wang
authors.forename: Lin
authors.initials: L
authors.lastname: Qiu
authors.forename: Cheli
authors.initials: C
authors.lastname: Wang
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jingyan
authors.initials: J
authors.lastname: Li
authors.forename: Jiang
authors.initials: J
authors.lastname: Xia
authors.forename: Pengju
authors.initials: P
authors.lastname: Jiang
mesh_terms: D000818:Animals; D000936:Antigen-Antibody Complex / Q000737:chemistry*; D000941:Antigens / Q000276:immunology*; D019075:Electrophoresis, Capillary; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D006041:Goats; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology*; D045663:Quantum Dots / Q000737:chemistry
chemical_list: D000936:Antigen-Antibody Complex; D000941:Antigens; D005456:Fluorescent Dyes; D007074:Immunoglobulin G
references.citation: Science. 2005 Jan 28;307(5709):538-44
references.pmid: 15681376
references.citation: Bioconjug Chem. 2007 Mar-Apr;18(2):323-32
references.pmid: 17373766
references.citation: Nat Protoc. 2008;3(1):89-96
references.pmid: 18193025
references.citation: J Phys Chem B. 2006 Mar 23;110(11):5779-89
references.pmid: 16539525
references.citation: Nat Protoc. 2006;1(3):1258-66
references.pmid: 17406409
references.citation: Anal Biochem. 1988 Oct;174(1):101-20
references.pmid: 3218725
references.citation: J Immunol Methods. 1997 Jan 15;200(1-2):155-9
references.pmid: 9005954
references.citation: Science. 2000 Jun 16;288(5473):2048-51
references.pmid: 10856219
references.citation: J Am Chem Soc. 2008 Mar 26;130(12):3744-5
references.pmid: 18321112
references.citation: Angew Chem Int Ed Engl. 2008;47(40):7602-25
references.pmid: 18810756
references.citation: Nano Lett. 2010 Jun 9;10(6):2192-6
references.pmid: 20481436
references.citation: Nat Biotechnol. 2004 Aug;22(8):969-76
references.pmid: 15258594
references.citation: J Am Chem Soc. 2006 Apr 26;128(16):5324-5
references.pmid: 16620087
references.citation: ACS Nano. 2012 Dec 21;6(12):11080-7
references.pmid: 23181687
references.citation: Angew Chem Int Ed Engl. 2012 Oct 15;51(42):10523-7
references.pmid: 22996637
references.citation: Methods. 1999 Oct;19(2):213-21
references.pmid: 10527727
references.citation: Anal Chim Acta. 2010 Jul 12;673(1):1-25
references.pmid: 20630173
references.citation: Phytochem Anal. 2004 Nov-Dec;15(6):365-71
references.pmid: 15595451
references.citation: Science. 1998 Sep 25;281(5385):2016-8
references.pmid: 9748158
references.citation: Nat Biotechnol. 2003 Jan;21(1):41-6
references.pmid: 12459735
references.citation: Biochemistry. 1995 May 16;34(19):6475-87
references.pmid: 7756279
references.citation: Biosens Bioelectron. 2010 Feb 15;25(6):1283-9
references.pmid: 19914053
references.citation: Biosens Bioelectron. 2009 Jan 1;24(5):1189-94
references.pmid: 18722763
references.citation: Chemistry. 2008;14(12):3637-44
references.pmid: 18318025
references.citation: ACS Nano. 2009 Apr 28;3(4):915-23
references.pmid: 19275175
references.citation: Science. 1998 Sep 25;281(5385):2013-6
references.pmid: 9748157
references.citation: Electrophoresis. 2012 Jul;33(13):1987-95
references.pmid: 22806464
references.citation: Nat Mater. 2005 Jun;4(6):435-46
references.pmid: 15928695