Abstract
Immunoassays have traditionally relied on antibodies as diagnostic probes. Their use outside of a laboratory, however, may be problematic because antibodies are often unstable in severe environmental conditions. Environmental monitoring requires thermostable probes, such as landscape phage, that carry thousands of foreign peptides on their surfaces, are superior to antibodies, and can operate in non-controlled conditions. While parent wild-type phage are known to be extremely stable in various media at high temperatures, no work has been done to demonstrate the stability of landscape phage probes. We examined the thermostability of a landscape phage probe and a monoclonal antibody specific for β-galactosidase in parallel in an enzyme-linked immunosorbent assay (ELISA) format. They were both stable for greater than six months at room temperature, but at higher temperatures the antibody degraded more rapidly than the phage probe. Phage retained detectable binding ability for more than six weeks at 63 °C, and three days at 76 °C. The activation energy of phage degradation was determined to be 1.34×105 J/mol. These results confirm that phage probes are highly thermostable and can function even after exposure to high temperatures during shipping, storage and operation.
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Acknowledgements
This work was supported by ARO/DARPA Grant # DAAD 19-01-10454 (to VAP) and NIH Grant # NIH-1 R21 AI055645 (to VAP). The authors thank Dr. Vitaly Vodyanoy for assistance with data analysis, and Director of AUDFS Center Dr. Bryan Chin for fruitful discussions and technical support of this work.
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Brigati, J.R., Petrenko, V.A. Thermostability of landscape phage probes. Anal Bioanal Chem 382, 1346–1350 (2005). https://doi.org/10.1007/s00216-005-3289-y
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DOI: https://doi.org/10.1007/s00216-005-3289-y