Reference - Detail
| RRC ID | 29141 |
|---|---|
| Author | Yokozeki T, Ueda H, Arai R, Mahoney W, Nagamune T. |
| Title | A homogeneous noncompetitive immunoassay for the detection of small haptens. |
| Journal | Anal Chem |
| Abstract |
We describe a noncompetitive homogeneous immunoassay for small haptens based on the antigen-dependent reassociation of antibody variable domains and beta-galactosidase (beta-gal) complementation (open sandwich enzymatic complementation immunoassay). As a model system, the reassociation of two fusion proteins, an anti 4-hydroxy-3-nitrophenylacetyl (NP) antibody heavy-chain variable-region fragment fused to an N-terminal deletion mutant of beta-gal (V(H)delta alpha) and the light-chain variable-region fragment fused to a C-terminal deletion mutant of beta-gal (V(L)delta omega), was monitored by the enzymatic complementation between the two. Upon simple mixing of the reagents with the sample, an antigen (NP)-dependent increase in enzymatic activity was observed. When 5-iodo-NP was measured, a 10 times higher sensitivity was observed, probably due to its higher affinity. Compared with our corresponding heterogeneous open sandwich enzyme-linked immunosorbent assay, approximately 1000-fold improvement in the sensitivity was attained, probably due to lower background V(H)-V(L) association. In addition, the assay required less time, handling, sample volume, and assay reagents. |
| Volume | 74(11) |
| Pages | 2500-4 |
| Published | 2002-6-1 |
| DOI | 10.1021/ac015743x |
| PMID | 12069229 |
| MeSH | Electrophoresis, Polyacrylamide Gel Escherichia coli / chemistry Escherichia coli / genetics Haptens / analysis* Immunoenzyme Techniques / methods* Plasmids / chemistry Recombinant Fusion Proteins / analysis beta-Galactosidase / chemistry |
| IF | 6.785 |
| Times Cited | 29 |
| WOS Category | CHEMISTRY, ANALYTICAL |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 12 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| DNA material | pPCR/delta alpha (RDB02987) pPCR/delta omega (RDB02988) |