{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T03:20:39Z","timestamp":1790306439370,"version":"4.1.0"},"reference-count":32,"publisher":"American Chemical Society (ACS)","issue":"19","license":[{"start":{"date-parts":[[2020,9,3]],"date-time":"2020-09-03T00:00:00Z","timestamp":1599091200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2020,9,3]],"date-time":"2020-09-03T00:00:00Z","timestamp":1599091200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2020,9,3]],"date-time":"2020-09-03T00:00:00Z","timestamp":1599091200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/100000165","name":"Division of Chemistry","doi-asserted-by":"publisher","award":["1608550"],"award-info":[{"award-number":["1608550"]}],"id":[{"id":"10.13039\/100000165","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,10,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Small-molecule detection in an immunoassay format generally employs competition or labeling. A novel direct-detection label-free primary immunoassay utilizing second harmonic generation (SHG) has been developed and the utility of the method has been demonstrated for several small-molecule narcotics. Specifically, the binding of morphine, methadone, and cocaine to antimorphine, antimethadone, and anticocaine antibodies was measured by SHG, allowing binding affinities and rates of dissociation to be obtained. The SHG primary immunoassay has provided the first kinetic measurements of small-molecule hapten interactions with a receptor antibody. The kinetics reveal for the first time that competitive immunoassays achieve their selectivity by taking advantage of the kinetics of association and dissociation of the labeled and unlabeled target and nontarget small-molecule to the capture antibody. In particular, the induced fit of the target small-molecule to their antibody pairs prolongs their residence time, while the nontarget small-molecule dissociate rapidly in comparison.<\/jats:p>","DOI":"10.1021\/acs.analchem.0c02286","type":"journal-article","created":{"date-parts":[[2020,9,3]],"date-time":"2020-09-03T02:48:53Z","timestamp":1599101333000},"page":"13163-13171","source":"Crossref","is-referenced-by-count":7,"title":["Revealing the Kinetic Advantage of a Competitive Small-Molecule\nImmunoassay by Direct Detection"],"prefix":"10.1021","volume":"92","author":[{"given":"Renee J.","family":"Tran","sequence":"first","affiliation":[{"name":"University of Utah , , 315 South 1400 East RM. 2020 , , ,","place":["Salt Lake City, Utah, United States, 84112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Krystal L.","family":"Sly","sequence":"additional","affiliation":[{"name":"University of Utah , , 315 South 1400 East RM. 2020 , , ,","place":["Salt Lake City, Utah, United States, 84112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0023-642X","authenticated-orcid":true,"given":"John C.","family":"Conboy","sequence":"additional","affiliation":[{"name":"University of Utah , , 315 South 1400 East RM. 2020 , , ,","place":["Salt Lake City, Utah, United States, 84112"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2020,9,3]]},"reference":[{"key":"2026081803312176000_cit1","doi-asserted-by":"publisher","first-page":"34","DOI":"10.1016\/j.jim.2016.06.004","article-title":"A Novel Immunoassay\nfor Quantitative Drug Abuse Screening in Serum","volume":"436","author":"Schumacher","year":"2016","journal-title":"J. Immunol. Methods"},{"key":"2026081803312176000_cit2","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1038\/284014a0","article-title":"More Sensitive\nImmunoassays","volume":"284","author":"Ekins","year":"1980","journal-title":"Nature"},{"key":"2026081803312176000_cit3","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1016\/j.aca.2010.04.023","article-title":"Validation of an Analytical Method\nfor Analysis of Cannabinoids in Hair by Headspace Solid-Phase Microextraction\nand Gas Chromatography\u2013Ion Trap Tandem Mass Spectrometry","volume":"670","author":"Emi\u0301dio","year":"2010","journal-title":"Anal. Chim. Acta"},{"key":"2026081803312176000_cit4","doi-asserted-by":"publisher","first-page":"7323","DOI":"10.3390\/s100807323","article-title":"Small Molecule\nImmunosensing Using Surface Plasmon Resonance","volume":"10","author":"Mitchell","year":"2010","journal-title":"Sensors"},{"key":"2026081803312176000_cit5","doi-asserted-by":"publisher","first-page":"2477","DOI":"10.1021\/ja043935e","article-title":"Fluorescent Cocaine Probes: A Tool\nfor the Selection\nand Engineering of Therapeutic Antibodies","volume":"127","author":"Meijler","year":"2005","journal-title":"J.\nAm. Chem. Soc."},{"key":"2026081803312176000_cit6","doi-asserted-by":"publisher","first-page":"2637","DOI":"10.1021\/ac048379l","article-title":"One-Step Homogenous\nImmunoassay for Small Analytes","volume":"77","author":"Pulli","year":"2005","journal-title":"Anal. Chem."},{"key":"2026081803312176000_cit7","doi-asserted-by":"publisher","first-page":"13399","DOI":"10.1021\/la802097z","article-title":"Effect of Fluorescently Labeling\nProtein Probes on Kinetics of Protein-Ligand Reactions","volume":"24","author":"Sun","year":"2008","journal-title":"Langmuir"},{"key":"2026081803312176000_cit8","doi-asserted-by":"crossref","DOI":"10.5772\/29620","article-title":"Variability of Plasma Methadone\nConcentration in Opiate Dependent Receiving Methadone: A Personalised\nApproach Towards Optimizing Dose","volume-title":"Toxicity\nand Drug Testing","author":"Mohamad","year":"2012"},{"key":"2026081803312176000_cit9","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0040518","article-title":"Probing\nCocaine-Antibody Interactions in Buffer and\nHuman Serum","volume":"7","author":"Ramakrishnan","year":"2012","journal-title":"PLoS One"},{"key":"2026081803312176000_cit10","doi-asserted-by":"publisher","first-page":"500","DOI":"10.1093\/jat\/23.6.500","article-title":"Detection of Methadone,\nLAAM, and\nTheir Metabolites by Methadone Immunoassays","volume":"23","author":"Cheever","year":"1999","journal-title":"J. Anal. Toxicol."},{"key":"2026081803312176000_cit11","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1002\/biof.1107","article-title":"Endogenous\nMorphine-6-Glucuronide (M6G) Is Present in the Plasma of Patients:\nValidation of a Specific Anti-M6G Antibody for Clinical and Basic\nResearch","volume":"40","author":"Laux-Biehlmann","year":"2014","journal-title":"BioFactors"},{"key":"2026081803312176000_cit12","doi-asserted-by":"publisher","first-page":"1271","DOI":"10.1111\/j.1471-4159.2011.07502.x","article-title":"A Morphine\/Heroin\nVaccine with New Hapten Design Attenuates Behavioral Effects in Rats","volume":"119","author":"Li","year":"2011","journal-title":"J. Neurochem."},{"key":"2026081803312176000_cit13","doi-asserted-by":"publisher","first-page":"570","DOI":"10.1016\/j.jmb.2005.03.080","article-title":"Diversity in Hapten Recognition:\nStructural Study of an Anti-Cocaine Antibody M82G2","volume":"349","author":"Pozharski","year":"2005","journal-title":"J. Mol. Biol."},{"key":"2026081803312176000_cit14","doi-asserted-by":"publisher","first-page":"691","DOI":"10.1016\/j.jmb.2003.12.084","article-title":"Anchoring a Cationic\nLigand: The\nStructure of the Fab Fragment of the Anti-Morphine Antibody 9B1 and\nIts Complex with Morphine","volume":"337","author":"Pozharski","year":"2004","journal-title":"J. Mol. Biol."},{"key":"2026081803312176000_cit15","volume-title":"The Principles of\nNonlinear Optics","author":"Shen","year":"1984"},{"key":"2026081803312176000_cit16","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1117\/12.238808","article-title":"Surface Second Harmonic Generation\n(SSHG) - A New Scheme for Immunoassay","volume":"2676","author":"Yang","year":"1996","journal-title":"Proc.\nSPIE"},{"key":"2026081803312176000_cit17","doi-asserted-by":"publisher","first-page":"221","DOI":"10.1016\/S0956-5663(00)00134-2","article-title":"An Immunosensor\nwith Potential for\nthe Detection of Viral Antigens in Body Fluids, Based on Surface Second\nHarmonic Generation","volume":"16","author":"Ditcham","year":"2001","journal-title":"Biosens. Bioelectron."},{"key":"2026081803312176000_cit18","doi-asserted-by":"publisher","first-page":"17290","DOI":"10.1073\/pnas.1905516116","article-title":"Second Harmonic\nGeneration Detection of Ras Conformational Changes and Discovery of\na Small Molecule Binder","volume":"116","author":"Donohue","year":"2019","journal-title":"Proc. Natl. Acad. Sci.\nU.S.A."},{"key":"2026081803312176000_cit19","doi-asserted-by":"publisher","first-page":"50","DOI":"10.1016\/j.jim.2016.01.001","article-title":"Considerations\nin Producing Preferentially Reduced Half-Antibody Fragments","volume":"429","author":"Makaraviciute","year":"2016","journal-title":"J. Immunol. Methods"},{"key":"2026081803312176000_cit20","doi-asserted-by":"publisher","first-page":"1252","DOI":"10.1016\/j.bios.2005.05.010","article-title":"DNA and\nProtein Microarray Printing\non Silicon Nitride Waveguide Surfaces","volume":"21","author":"Wu","year":"2006","journal-title":"Biosens.\nBioelectron."},{"key":"2026081803312176000_cit21","doi-asserted-by":"publisher","first-page":"1013","DOI":"10.1364\/JOSAB.21.001013","article-title":"Counterpropagating\nSecond-Harmonic Generation: A New\nTechnique for the Investigation of Molecular Chirality at Surfaces","volume":"21","author":"Kriech","year":"2004","journal-title":"J. Opt. Soc. Am. B"},{"key":"2026081803312176000_cit22","doi-asserted-by":"publisher","first-page":"201","DOI":"10.1021\/ac202375n","article-title":"Comparison of the Energetics of Avidin,\nStreptavidin, NeutrAvidin, and Anti-Biotin Antibody Binding to Biotinylated\nLipid Bilayer Examined by Second-Harmonic Generation","volume":"84","author":"Nguyen","year":"2012","journal-title":"Anal. Chem."},{"key":"2026081803312176000_cit23","doi-asserted-by":"publisher","DOI":"10.3389\/fneur.2015.00179","article-title":"A Practical Guide to Immunoassay\nMethod Validation","volume":"6","author":"Andreasson","year":"2015","journal-title":"Front. Neurol."},{"key":"2026081803312176000_cit24","volume-title":"Chemical Kinetics","author":"Laidler","year":"1987","edition":"3"},{"key":"2026081803312176000_cit25","doi-asserted-by":"publisher","first-page":"1954","DOI":"10.1021\/ac061288z","article-title":"Least Detectable Concentration\nand\nDynamic Range of Three Immunoassay Systems Using the Same Antibody","volume":"79","author":"Glass","year":"2007","journal-title":"Anal. Chem."},{"key":"2026081803312176000_cit26","first-page":"129","article-title":"Study of\na Mathematical Theory for the Coagulation Kinetics of Colloidal Solutions","volume":"92","author":"Smoluchowski","year":"1917","journal-title":"Z. Phys. Chem."},{"key":"2026081803312176000_cit27","doi-asserted-by":"publisher","first-page":"12730","DOI":"10.1073\/pnas.0500909102","article-title":"Structure and Kinetics of a Transient Antibody Binding\nIntermediate Reveal a Kinetic Discrimination Mechanism in Antigen\nRecognition","volume":"102","author":"James","year":"2005","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026081803312176000_cit28","doi-asserted-by":"publisher","first-page":"1383","DOI":"10.1016\/j.addr.2008.04.010","article-title":"The Chemistry\nof Irreversible Capture","volume":"60","author":"Meares","year":"2008","journal-title":"Adv. Drug Delivery\nRev."},{"key":"2026081803312176000_cit29","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1093\/jb\/mvw050","article-title":"Role of the Mobility\nof Antigen Binding\nSite in High Affinity Antibody Elucidated by Surface Plasmon Resonance","volume":"161","author":"Fukuda","year":"2017","journal-title":"J. Biochem."},{"key":"2026081803312176000_cit30","doi-asserted-by":"publisher","first-page":"887","DOI":"10.1016\/j.sbi.2013.06.014","article-title":"Modeling Protein\nAssociation Mechanism and Kinetics","volume":"23","author":"Zhou","year":"2013","journal-title":"Curr. Opin.\nStruct. Biol."},{"key":"2026081803312176000_cit31","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1016\/S0006-3495(82)84455-X","article-title":"Role of Diffusion in\nLigand Binding to Macromolecules and Cell-Bound\nReceptors","volume":"40","author":"Shoup","year":"1982","journal-title":"Biophys. J."},{"key":"2026081803312176000_cit32","doi-asserted-by":"publisher","first-page":"5979","DOI":"10.1021\/ac2009614","article-title":"High-Throughput Screening of Drug-Lipid\nMembrane Interactions\nvia Counter-Propagating Second Harmonic Generation Imaging","volume":"83","author":"Nguyen","year":"2011","journal-title":"Anal. Chem."}],"container-title":["Analytical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.analchem.0c02286","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/92\/19\/13163\/8592288\/ac0c02286.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/92\/19\/13163\/8592288\/ac0c02286.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T08:59:52Z","timestamp":1787043592000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/ancham\/article\/92\/19\/13163\/828171\/Revealing-the-Kinetic-Advantage-of-a-Competitive"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,9,3]]},"references-count":32,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2020,9,3]]},"published-print":{"date-parts":[[2020,10,6]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.analchem.0c02286","relation":{},"ISSN":["0003-2700","1520-6882"],"issn-type":[{"value":"0003-2700","type":"print"},{"value":"1520-6882","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,9,3]]}}}