Simultaneous Detection of Multiple Proteins with an Array-Based Enzyme-Linked Immunosorbent Assay (ELISA) and Enhanced Chemiluminescence (ECL)
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Ruo-Pan Huang
Abstract
Protein arrays hold a promise in basic and clinical applications. As the first step to develop such array system, I used an array-based enzyme-linked immunosorbent assay (ELISA) and enhanced chemiluminescence (ECL) to demonstrate the feasibility of simultaneous detection of multiple proteins. In the direct ELISA system, different known immunoglobulin Gs (IgGs) were immobilized onto polyvinylidine difloride (PDVF) membrane through 96-well format Bio-Dot unit. The antigens were then individually and collectively detected by incubation of membranes with different antibodies coupled with ECL. In the sandwich ELISA system, the cytokine capture antibodies were immobilized onto PDVF membranes. The membranes were then incubated with single cytokine or a combination of different cytokines. The captured cytokines were detected by biotin-conjugated antibodies coupled with ECL system. Experiments demonstrated that multiple IgGs and cytokines could be simultaneously detected using this approach with high specificity and sensitivity. More importantly, cytokines from biological samples were detected using this approach, which can be used in any general laboratory setting without any sophisticated equipment. This concept could be extended to develop a protein-based array system.
Copyright © 2001 by Walter de Gruyter GmbH & Co. KG
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- Hippuric Acid as a Modifier of Calcium Oxalate Crystallisation
- Excretion of Sweat and Urine Pyridinoline Crosslinks in Healthy Controls and Subjects with Established Metabolic Bone Disease
- Hyperhomocysteinemia and Changed Plasma Thiol Redox Status in Chronic Obstructive Pulmonary Disease
- Biochemical Evaluation of Oxidative Stress during Exercise in Patients with Coronary Heart Disease
- Prognostic Significance of the Presence of Erythroblasts in Blood after Cardiothoracic Surgery
- Exploratory Biochemical Data Analysis: a Comparison of Two Sample Means and Diagnostic Displays
- Standardization of Laboratory Data and Establishment of Reference Intervals in the Fukuoka Prefecture: A Japanese Perspective
- Calibration, Specificity and Trueness of a Postheparin Plasma Lipoprotein Lipase Assay
- Analytical Performance of a Direct Assay for LDL-Cholesterol
- Genotyping Method for Point Mutation Detection in the Endothelial Nitric Oxide Synthase Exon 7 Using Fluorescent Probes. Clinical Validation in Systemic Sclerosis Patients
- IFCC Reference Measurement Procedure for Substance Concentration Determination of Total Carbon Dioxide in Blood, Plasma or Serum
- Joshua Lederberg (Editor-in-Chief): Encyclopedia of Microbiology, 2nd Edition, Vol 1-4
Articles in the same Issue
- Autoantibodies Associated with Rheumatic Diseases
- Simultaneous Detection of Multiple Proteins with an Array-Based Enzyme-Linked Immunosorbent Assay (ELISA) and Enhanced Chemiluminescence (ECL)
- Hippuric Acid Test Using 13C-Labelling and NMR Spectroscopy
- Hippuric Acid as a Modifier of Calcium Oxalate Crystallisation
- Excretion of Sweat and Urine Pyridinoline Crosslinks in Healthy Controls and Subjects with Established Metabolic Bone Disease
- Hyperhomocysteinemia and Changed Plasma Thiol Redox Status in Chronic Obstructive Pulmonary Disease
- Biochemical Evaluation of Oxidative Stress during Exercise in Patients with Coronary Heart Disease
- Prognostic Significance of the Presence of Erythroblasts in Blood after Cardiothoracic Surgery
- Exploratory Biochemical Data Analysis: a Comparison of Two Sample Means and Diagnostic Displays
- Standardization of Laboratory Data and Establishment of Reference Intervals in the Fukuoka Prefecture: A Japanese Perspective
- Calibration, Specificity and Trueness of a Postheparin Plasma Lipoprotein Lipase Assay
- Analytical Performance of a Direct Assay for LDL-Cholesterol
- Genotyping Method for Point Mutation Detection in the Endothelial Nitric Oxide Synthase Exon 7 Using Fluorescent Probes. Clinical Validation in Systemic Sclerosis Patients
- IFCC Reference Measurement Procedure for Substance Concentration Determination of Total Carbon Dioxide in Blood, Plasma or Serum
- Joshua Lederberg (Editor-in-Chief): Encyclopedia of Microbiology, 2nd Edition, Vol 1-4
