title: ELISA in the multiplex era: potentials and pitfalls.
abstract: Multiplex immunoassays confer several advantages over widely adopted singleplex immunoassays including increased efficiency at a reduced expense, greater output per sample volume ratios and higher throughput predicating more resolute, detailed diagnostics and facilitating personalised medicine. Nonetheless, to date, relatively few protein multiplex immunoassays have been validated for in vitro diagnostics in clinical/point-of-care settings. This review article will outline the challenges, which must be ameliorated prior to the widespread integration of multiplex immunoassays in clinical settings: (i) biomarker validation; (ii) standardisation of immunoassay design and quality control (calibration and quantification); (iii) availability, stability, specificity and cross-reactivity of reagents; (iv) assay automation and the use of validated algorithms for transformation of raw data into diagnostic results. A compendium of multiplex immunoassays applicable to in vitro diagnostics and a summary of the diagnostic products currently available commercially are included, along with an analysis of the relative states of development for each format (namely planar slide based, suspension and planar/microtitre plate based) with respect to the aforementioned issues.
authors: School of Life Sciences, The University of Nottingham, Nottingham, UK.
authors: Patrick J
authors: PJ
authors: Tighe
authors: Richard R
authors: RR
authors: Ryder
authors: Ian
authors: I
authors: Todd
authors: Lucy C
authors: LC
authors: Fairclough
mesh_terms: D015415:Biomarkers / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007118:Immunoassay / Q000379:methods*
chemical_list: D015415:Biomarkers
keywords: ELISA; In vitro diagnostics; Multiplex immunoassay
references: J Immunol Methods. 2000 Feb 21;235(1-2):71-80
references: 10675759
references: Biotechniques. 2001 Jul;31(1):186-90, 192-4
references: 11464511
references: Nature. 1975 Aug 7;256(5517):495-7
references: 1172191
references: J Immunol Methods. 1992 Jul 6;151(1-2):47-66
references: 1378475
references: J Clin Pathol. 1963 Jan;16:282-4
references: 13962737
references: J Immunol Methods. 2005 May;300(1-2):124-35
references: 15896801
references: J Immunol Methods. 2006 Feb 20;309(1-2):205-8
references: 16436279
references: Methods. 2006 Apr;38(4):294-303
references: 16481197
references: Cytometry A. 2006 May;69(5):318-25
references: 16604537
references: Drug Discov Today. 2006 Nov;11(21-22):1007-11
references: 17055410
references: J Allergy Clin Immunol. 2007 Feb;119(2):428-33
references: 17196246
references: Science. 2007 May 18;316(5827):1039-43
references: 17463250
references: Proteomics. 2007 Aug;7(16):2738-50
references: 17639606
references: Science. 2007 Oct 12;318(5848):287-90
references: 17872411
references: J Biol Chem. 2008 Jan 11;283(2):739-50
references: 17942395
references: Am J Clin Pathol. 2008 May;129(5):788-95
references: 18426740
references: Anal Bioanal Chem. 2009 Mar;393(5):1407-16
references: 18807017
references: Clin Cancer Res. 2008 Oct 1;14(19):5967-76
references: 18829475
references: J Immunol Methods. 2009 Jan 1;340(1):55-64
references: 18983846
references: J Immunol Methods. 2009 Mar 15;342(1-2):106-14
references: 19174166
references: Clin Chem. 2009 Jun;55(6):1092-9
references: 19372187
references: J Mol Diagn. 2009 Nov;11(6):590-7
references: 19767590
references: Clin Chem. 2010 Feb;56(2):186-93
references: 19959625
references: Clin Chem. 2010 Feb;56(2):314-8
references: 20022982
references: Br J Cancer. 2010 May 11;102(10):1524-32
references: 20407440
references: Clin Vaccine Immunol. 2010 Jun;17(6):966-8
references: 20427627
references: Alzheimers Dement. 2010 May;6(3):230-8
references: 20451871
references: Cancer Cell Int. 2010 Nov 24;10:45
references: 21092330
references: Hear Res. 2011 May;275(1-2):1-7
references: 21144888
references: Microbiol Immunol. 2011 Apr;55(4):267-78
references: 21244466
references: J Immunol Methods. 2011 Mar 31;367(1-2):33-9
references: 21277854
references: Clin Chem. 2011 Apr;57(4):560-7
references: 21300740
references: Korean J Lab Med. 2011 Apr;31(2):61-71
references: 21474978
references: J Pharm Biomed Anal. 2011 Jul 15;55(5):869-77
references: 21530130
references: Malar J. 2011 May 09;10:115
references: 21554671
references: J Obstet Gynaecol. 2011 Jul;31(5):375-9
references: 21627416
references: Hum Reprod. 2011 Sep;26(9):2289-95
references: 21685139
references: J Oral Maxillofac Surg. 2012 May;70(5):1070-80
references: 21802820
references: Nutr Hosp. 2011 Sep-Oct;26(5):1115-9
references: 22072361
references: Mol Cell Proteomics. 2012 Apr;11(4):M111.011460
references: 22171321
references: Genome Res. 2012 Feb;22(2):183-7
references: 22301132
references: Sensors (Basel). 2012;12(2):1494-508
references: 22438722
references: J Otolaryngol Head Neck Surg. 2012 Feb;41(1):51-7
references: 22498269
references: Nat Rev Cardiol. 2012 May 01;9(8):478-90
references: 22547172
references: Clin Cancer Res. 2012 May 1;18(9):2687-94
references: 22553374
references: Pharmacol Rep. 2012;64(1):179-84
references: 22580534
references: J Neuroimmunol. 2012 Aug 15;249(1-2):109-11
references: 22633195
references: Arch Physiol Biochem. 2012 Oct;118(4):192-6
references: 22852821
references: Arthritis Res Ther. 2012 Oct 01;14(5):R201
references: 23025688
references: PLoS One. 2012;7(9):e44260
references: 23028511
references: J Immunol Methods. 2013 Jan 31;387(1-2):89-95
references: 23085532
references: Nephrol Dial Transplant. 2013 Feb;28(2):254-73
references: 23115326
references: Clin Transl Allergy. 2012 Dec 19;2(1):24
references: 23254184
references: PLoS One. 2012;7(12):e52981
references: 23285240
references: PLoS One. 2013;8(1):e52587
references: 23300981
references: Methods. 2013 May 15;61(1):23-9
references: 23333412
references: BMC Bioinformatics. 2013 Apr 30;14:145
references: 23631706
references: Bioanalysis. 2013 May;5(9):1041-55
references: 23641695
references: Vaccine. 2013 Jul 18;31(33):3333-8
references: 23742991
references: Best Pract Res Clin Endocrinol Metab. 2013 Oct;27(5):647-61
references: 24094636
references: Pediatr Allergy Immunol. 2014 Feb;25(1):71-9
references: 24131308
references: J Clin Microbiol. 2014 Jan;52(1):103-7
references: 24153130
references: J Immunol Methods. 2013 Dec 31;400-401:58-69
references: 24161315
references: PLoS One. 2013 Nov 04;8(11):e79662
references: 24223987
references: Int Arch Occup Environ Health. 2014;87(7):775-81
references: 24310757
references: Analyst. 2014 Feb 7;139(3):528-42
references: 24326290
references: Int J Biol Sci. 2013 Nov 22;9(10):1099-107
references: 24339731
references: Cytotherapy. 2014 May;16(5):631-9
references: 24364910
references: Results Immunol. 2011 Dec 17;2:1-6
references: 24371560
references: Arthritis Res Ther. 2014 Jan 08;16(1):R5
references: 24398122
references: J Immunol Methods. 2014 Mar;405:29-34
references: 24424297
references: Clin Vaccine Immunol. 2014 Apr;21(4):496-500
references: 24477853
references: Arthritis Res Ther. 2013 Nov 07;15(6):R182
references: 24485167
references: PLoS One. 2014 Feb 06;9(2):e88394
references: 24516646
references: Curr Opin Chem Biol. 2014 Feb;18:29-37
references: 24534750
references: Biol Psychiatry. 2014 May 1;75(9):723-31
references: 24548642
references: J Neurol Sci. 2014 May 15;340(1-2):75-9
references: 24629476
references: PLoS One. 2014 Mar 25;9(3):e93144
references: 24667736
references: Clin Transl Allergy. 2014 Mar 28;4(1):9
references: 24679244
references: BMC Biotechnol. 2014 Apr 01;14:24
references: 24684904
references: PLoS Negl Trop Dis. 2014 Apr 10;8(4):e2786
references: 24722524
references: Prev Med. 2014 Jul;64:81-7
references: 24732722
references: J Immunol Methods. 2014 Jun;408:13-23
references: 24768796
references: Sci Rep. 2014 May 02;4:4878
references: 24786974
references: J Pharm Biomed Anal. 1989;7(2):155-68
references: 2488616
references: Analyst. 2014 Oct 21;139(20):5093-102
references: 25112421
references: Ann Dermatol. 2014 Aug;26(4):437-46
references: 25143671
references: Clin Chem. 1988 Jan;34(1):27-33
references: 3338181
references: J Immunol Methods. 1983 Dec 16;65(1-2):217-23
references: 6606681
references: Cancer Res. 1982 Mar;42(3):1088-93
references: 7059971
references: Science. 1977 Oct 14;198(4313):149-57
references: 905822
references: Clin Chem. 1999 Feb;45(2):199-205
references: 9931041