Thermostability of landscape phage probes

被引:83
作者
Brigati, JR [1 ]
Petrenko, VA [1 ]
机构
[1] Auburn Univ, Coll Vet Med, Dept Pathobiol, Auburn, AL 36849 USA
关键词
landscape phage; diagnostic probes; thermostability; beta-galactosidase; enzyme-linked immunosorbent assay (ELISA);
D O I
10.1007/s00216-005-3289-y
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 beta-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 degrees C, and three days at 76 degrees C. The activation energy of phage degradation was determined to be 1.34x10(5) 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.
引用
收藏
页码:1346 / 1350
页数:5
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