Competitive Immunoassays for the Detection of Small Molecules Using Single Molecule Arrays
Small-molecule detection is important for many applications including clinical diagnostics, drug discovery, and measurements of environmental samples and agricultural products. Current techniques for small-molecule detection suffer from various limitations including low analytical sensitivity and co...
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| Published in | Journal of the American Chemical Society Vol. 140; no. 51; pp. 18132 - 18139 |
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| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
American Chemical Society
26.12.2018
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| Subjects | |
| Online Access | Get full text |
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| Abstract | Small-molecule detection is important for many applications including clinical diagnostics, drug discovery, and measurements of environmental samples and agricultural products. Current techniques for small-molecule detection suffer from various limitations including low analytical sensitivity and complex sample processing. Furthermore, as a result of their small size, small molecules are difficult to detect using an antibody pair in a traditional sandwich assay format. To overcome these limitations, we developed an ultrasensitive competitive immunoassay for small-molecule detection using Single Molecule Arrays (Simoa). We show that the competitive Simoa assay is approximately 50-fold more sensitive than the conventional ELISA. We performed theoretical calculations to determine the factors that influence the sensitivity of competitive Simoa assays and used them to achieve maximal sensitivity. We also demonstrate detection of small molecules in complex biological samples. We show that the competitive Simoa assay is a simple, fast, and highly sensitive approach for ultrasensitive detection of small molecules. |
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| AbstractList | Small-molecule detection is important for many applications including clinical diagnostics, drug discovery, and measurements of environmental samples and agricultural products. Current techniques for small-molecule detection suffer from various limitations including low analytical sensitivity and complex sample processing. Furthermore, as a result of their small size, small molecules are difficult to detect using an antibody pair in a traditional sandwich assay format. To overcome these limitations, we developed an ultrasensitive competitive immunoassay for small-molecule detection using Single Molecule Arrays (Simoa). We show that the competitive Simoa assay is approximately 50-fold more sensitive than the conventional ELISA. We performed theoretical calculations to determine the factors that influence the sensitivity of competitive Simoa assays and used them to achieve maximal sensitivity. We also demonstrate detection of small molecules in complex biological samples. We show that the competitive Simoa assay is a simple, fast, and highly sensitive approach for ultrasensitive detection of small molecules. Small-molecule detection is important for many applications including clinical diagnostics, drug discovery, and measurements of environmental samples and agricultural products. Current techniques for small-molecule detection suffer from various limitations including low analytical sensitivity and complex sample processing. Furthermore, as a result of their small size, small molecules are difficult to detect using an antibody pair in a traditional sandwich assay format. To overcome these limitations, we developed an ultrasensitive competitive immunoassay for small-molecule detection using Single Molecule Arrays (Simoa). We show that the competitive Simoa assay is approximately 50-fold more sensitive than the conventional ELISA. We performed theoretical calculations to determine the factors that influence the sensitivity of competitive Simoa assays and used them to achieve maximal sensitivity. We also demonstrate detection of small molecules in complex biological samples. We show that the competitive Simoa assay is a simple, fast, and highly sensitive approach for ultrasensitive detection of small molecules.Small-molecule detection is important for many applications including clinical diagnostics, drug discovery, and measurements of environmental samples and agricultural products. Current techniques for small-molecule detection suffer from various limitations including low analytical sensitivity and complex sample processing. Furthermore, as a result of their small size, small molecules are difficult to detect using an antibody pair in a traditional sandwich assay format. To overcome these limitations, we developed an ultrasensitive competitive immunoassay for small-molecule detection using Single Molecule Arrays (Simoa). We show that the competitive Simoa assay is approximately 50-fold more sensitive than the conventional ELISA. We performed theoretical calculations to determine the factors that influence the sensitivity of competitive Simoa assays and used them to achieve maximal sensitivity. We also demonstrate detection of small molecules in complex biological samples. We show that the competitive Simoa assay is a simple, fast, and highly sensitive approach for ultrasensitive detection of small molecules. |
| Author | Walt, David R Wang, Xu Wang, Jun Cohen, Limor |
| AuthorAffiliation | Department of Pathology, Brigham and Women’s Hospital Department of Chemical Biology School of Physical and Mathematical Sciences Wyss Institute for Biologically Inspired Engineering |
| AuthorAffiliation_xml | – name: Department of Chemical Biology – name: Wyss Institute for Biologically Inspired Engineering – name: Department of Pathology, Brigham and Women’s Hospital – name: School of Physical and Mathematical Sciences |
| Author_xml | – sequence: 1 givenname: Xu orcidid: 0000-0002-5929-5048 surname: Wang fullname: Wang, Xu organization: Department of Pathology, Brigham and Women’s Hospital – sequence: 2 givenname: Limor orcidid: 0000-0003-1448-0925 surname: Cohen fullname: Cohen, Limor organization: Department of Pathology, Brigham and Women’s Hospital – sequence: 3 givenname: Jun surname: Wang fullname: Wang, Jun organization: School of Physical and Mathematical Sciences – sequence: 4 givenname: David R orcidid: 0000-0002-5524-7348 surname: Walt fullname: Walt, David R email: dwalt@bwh.harvard.edu organization: Department of Pathology, Brigham and Women’s Hospital |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/30495929$$D View this record in MEDLINE/PubMed |
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| SubjectTerms | agricultural products antibodies detection limit diagnostic techniques drugs enzyme-linked immunosorbent assay |
| Title | Competitive Immunoassays for the Detection of Small Molecules Using Single Molecule Arrays |
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