A Near-Infrared, Surface-Enhanced, Fluorophore-Linked Immunosorbent Assay
The enzyme-linked immunosorbent assay is commonly used for research and clinical applications but typically suffers from a limited linear range and is difficult to multiplex. The fluorophore-linked immunosorbent assay is a closely related technique with good linear range and the ability to detect mu...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2013-08, Vol.85 (15), p.7102-7108 |
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| creator | Furtaw, Michael D Steffens, David L Urlacher, Teresa M Anderson, Jon P |
| description | The enzyme-linked immunosorbent assay is commonly used for research and clinical applications but typically suffers from a limited linear range and is difficult to multiplex. The fluorophore-linked immunosorbent assay is a closely related technique with good linear range and the ability to detect multiple antigens simultaneously but is typically less sensitive. Here, we demonstrate a near-infrared, surface-enhanced fluorophore-linked immunosorbent assay with sensitivity comparable to its enzyme-linked counterpart. A 59-fold enhancement to sensitivity (slope of linear fit) and an 8-fold improvement in LOD are demonstrated on a direct assay with rabbit immunoglobulin-G as a model system. The technique is also tested on a clinically relevant assay to detect alpha-fetoprotein, in which a 42-fold enhancement to sensitivity is demonstrated along with a 16-fold improvement in LOD. The technique enables these accomplishments while maintaining the entire traditional assay protocol and simply adding two steps at the end. This technique may prove superior to current protocols for biomarker research and clinical diagnoses, which require high sensitivity along with quantitation over an extended range. |
| doi_str_mv | 10.1021/ac400536k |
| format | Article |
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Chem</addtitle><description>The enzyme-linked immunosorbent assay is commonly used for research and clinical applications but typically suffers from a limited linear range and is difficult to multiplex. The fluorophore-linked immunosorbent assay is a closely related technique with good linear range and the ability to detect multiple antigens simultaneously but is typically less sensitive. Here, we demonstrate a near-infrared, surface-enhanced fluorophore-linked immunosorbent assay with sensitivity comparable to its enzyme-linked counterpart. A 59-fold enhancement to sensitivity (slope of linear fit) and an 8-fold improvement in LOD are demonstrated on a direct assay with rabbit immunoglobulin-G as a model system. The technique is also tested on a clinically relevant assay to detect alpha-fetoprotein, in which a 42-fold enhancement to sensitivity is demonstrated along with a 16-fold improvement in LOD. 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| source | ACS Publications; MEDLINE |
| subjects | alpha-Fetoproteins - analysis Analytical chemistry Animals Antigens Assaying Benzenesulfonates - chemistry Biomarkers Enzymes Fluorescent Dyes - chemistry Immunoglobulin G - immunology Immunoglobulins Immunosorbent Techniques Indoles - chemistry Infrared Rays Multiplexing Rabbits Slopes Spectrometry, Fluorescence Surface Properties |
| title | A Near-Infrared, Surface-Enhanced, Fluorophore-Linked Immunosorbent Assay |
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