Development of a specific troponin I detection system with enhanced immune sensitivity using a single monoclonal antibody

Using an immunoassay in combination with surface plasmon fluorescence spectroscopy (SPFS), we report the rapid detection of troponin I, a valuable biomarker for diagnosis of myocardial infarction. We discuss the implementation of (i) direct, (ii) sandwich, and (iii) competitive assay formats, based on surface plasmon resonance and SPFS. To elucidate the results, we relate the experiments to orientation-dependent interaction of troponin I epitopes with respective immunoglobulin G antibodies. A limit of detection (LoD) of 19 pM, with 45 min readout time, was achieved using single monoclonal antibody that is specific for one epitope. The borderline between normal people and patients is 20 pM to 83 pM cTnI concentration, and upon the outbreak of acute myocardial infraction it can raise to 2 nM and levels at 20 nM for 6–8 days, therefore the achieved LoD covers most of the clinically relevant range. In addition, this system allows for the detection of troponin I using a single specific monoclonal antibody, which is highly beneficial in case of detection in real samples, where the protein has a complex form leading to hidden epitopes, thus paving the way towards a system that can improve early-stage screening of heart attacks.

[1]  W. Knoll,et al.  Compact Grating-Coupled Biosensor for the Analysis of Thrombin. , 2019, ACS sensors.

[2]  R. El-Kased.  Immuno-analytical approach and its application for cardiac disease marker detection , 2018, Journal of immunoassay & immunochemistry.

[3]  X. He,et al.  Rapid and Sensitive Detection of Cardiac Troponin I for Point-of-Care Tests Based on Red Fluorescent Microspheres , 2018, Molecules.

[4]  Shalini Prasad,et al.  Cardiac troponin biosensors: where are we now? , 2018 .

[5]  C. Rodriguez-Emmenegger,et al.  Plasmonic Hepatitis B Biosensor for the Analysis of Clinical Saliva , 2017, Analytical chemistry.

[6]  B. Liedberg,et al.  Peptide Functionalized Gold Nanoparticles with Optimized Particle Size and Concentration for Colorimetric Assay Development: Detection of Cardiac Troponin I , 2016 .

[7]  Lin Wu,et al.  Exploiting Surface-Plasmon-Enhanced Light Scattering for the Design of Ultrasensitive Biosensing Modality. , 2016, Analytical chemistry.

[8]  J. Dostálek,et al.  Plasmonically amplified bioassay - Total internal reflection fluorescence vs. epifluorescence geometry. , 2016, Talanta.

[9]  Roger M. Leblanc,et al.  Recent Development of Cardiac Troponin i Detection , 2016 .

[10]  Rajesh R. Naik,et al.  Peptide Functionalized Gold Nanorods for the Sensitive Detection of a Cardiac Biomarker Using Plasmonic Paper Devices , 2015, Scientific Reports.

[11]  Feng Xu,et al.  Biomarker detection for disease diagnosis using cost-effective microfluidic platforms. , 2015, The Analyst.

[12]  Yang Zhang,et al.  The I-TASSER Suite: protein structure and function prediction , 2014, Nature Methods.

[13]  Jakub Dostalek,et al.  Plasmon-Enhanced Fluorescence Biosensors: a Review , 2013, Plasmonics.

[14]  A. Bereznikova,et al.  Epitope specificity of anti-cardiac troponin I monoclonal antibody 8I-7. , 2013, Clinical chemistry.

[15]  Qingge Xu,et al.  The impact of antibody selection on the detection of cardiac troponin I. , 2013, Clinica chimica acta; international journal of clinical chemistry.

[16]  J. Niazi,et al.  Biosensors for cardiac biomarkers detection: A review , 2012 .

[17]  Tsehai A. J. Grell,et al.  Rapid and sensitive detection of troponin I in human whole blood samples by using silver nanoparticle films and microwave heating. , 2011, Clinical chemistry.

[18]  Yang Zhang,et al.  I-TASSER: a unified platform for automated protein structure and function prediction , 2010, Nature Protocols.

[19]  M. Fischer,et al.  Amine coupling through EDC/NHS: a practical approach. , 2010, Methods in molecular biology.

[20]  W. Knoll,et al.  Long range surface plasmon-enhanced fluorescence spectroscopy for the detection of aflatoxin M1 in milk. , 2009, Biosensors & bioelectronics.

[21]  Fred S Apple,et al.  Assessment of the multiple-biomarker approach for diagnosis of myocardial infarction in patients presenting with symptoms suggestive of acute coronary syndrome. , 2009, Clinical chemistry.

[22]  W. Knoll,et al.  New concepts with surface plasmons and nano-biointerfaces , 2008 .

[23]  Yang Zhang,et al.  I-TASSER server for protein 3D structure prediction , 2008, BMC Bioinformatics.

[24]  J. Alpert,et al.  Universal definition of myocardial infarction. , 2007, European heart journal.

[25]  Jeroen J. Bax,et al.  Universal definition of myocardial infarction. , 2007 .

[26]  J. Dostálek,et al.  Multichannel SPR biosensor for detection of endocrine-disrupting compounds , 2007, Analytical and bioanalytical chemistry.

[27]  Holger H. Hoos,et al.  A replica exchange Monte Carlo algorithm for protein folding in the HP model , 2007, BMC Bioinformatics.

[28]  W. Knoll,et al.  Long Range Surface Plasmons for Observation of Biomolecular Binding Events at Metallic Surfaces , 2007 .

[29]  Jean-Philippe Charrier,et al.  Isoelectric point determination of cardiac troponin I forms present in plasma from patients with myocardial infarction. , 2007, Clinica chimica acta; international journal of clinical chemistry.

[30]  Lars Edenbrandt,et al.  Direct hospital costs of chest pain patients attending the emergency department: a retrospective study , 2006, BMC emergency medicine.

[31]  D. Cook,et al.  Elevated cardiac troponin levels in critically ill patients: prevalence, incidence, and outcomes. , 2006, American journal of critical care : an official publication, American Association of Critical-Care Nurses.

[32]  M. Nusier,et al.  Diagnostic Efficiency of Creatine Kinase (CK), CKMB, Troponin T and Troponin I in Patients with Suspected Acute Myocardial Infarction , 2006 .

[33]  A. Jaffe,et al.  Troponin: the biomarker of choice for the detection of cardiac injury , 2005, Canadian Medical Association Journal.

[34]  A. Katrukha.  Antibody Selection Strategies in Cardiac Troponin Assays , 2003 .

[35]  A. Go,et al.  The prognostic value of troponin in patients with non-ST elevation acute coronary syndromes: a meta-analysis. , 2001, Journal of the American College of Cardiology.

[36]  Wolfgang Knoll,et al.  Surface-Plasmon Field-Enhanced Fluorescence Spectroscopy , 2000 .

[37]  W. Knoll,et al.  Interfaces and thin films as seen by bound electromagnetic waves. , 1998, Annual review of physical chemistry.

[38]  K. Pulkki,et al.  Troponin I is released in bloodstream of patients with acute myocardial infarction not in free form but as complex. , 1997, Clinical chemistry.

[39]  G. Burch.  [Cardiovascular diseases]. , 1956, Revista medica cubana.