Stabilization of Proteins and Peptides in Diagnostic Immunological Assays by the Molecular Chaperone Hsp25
| Autor: | Alfons Dr Nichtl, Monika Ehrnsperger, Johannes Buchner, Christoph Hergersberg, Ulla Wienhues |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Antigenicity
Protein Conformation Molecular Sequence Data Complex formation HSP27 Heat-Shock Proteins Biophysics Enzyme-Linked Immunosorbent Assay Viral Nonstructural Proteins Biology Biochemistry Troponin T Antigen Amino Acid Sequence Molecular Biology Small Heat-Shock Proteins Heat-Shock Proteins chemistry.chemical_classification Proteins alpha-Glucosidases Cell Biology Troponin Neoplasm Proteins Enzyme chemistry biology.protein Protein folding Antibody Peptides Molecular Chaperones |
| Zdroj: | Analytical Biochemistry. 259:218-225 |
| ISSN: | 0003-2697 |
| DOI: | 10.1006/abio.1998.2630 |
| Popis: | Diagnostic assays for proteins devoid of enzymatic activity are becoming increasingly important. Antibodies generated against the respective proteins are used for their detection in enzyme-linked immunosorbent assay or patient sera are used to monitor disease-related antibodies against recombinantly produced antigens. A problem frequently encountered with these assays is that the proteins or fragments thereof used as standards have a limited shelf life. A similar problem arises when activities of labile enzymes are used for diagnostic detection. Here, we present a novel approach to ‘stabilizing‘ enzymatic activity and antigenicity of proteins used for immunogenic detection by molecular chaperones. We have exploited the ability of molecular chaperones to keep proteins in their active conformation to overcome the biotechnological problems encountered in protein-based diagnostics of heart attack, stroke, and viral infections such as hepatitis C. We show that Hsp25, a member of the family of small heat shock proteins, known to act as a molecular chaperone in protein folding reactions, can stably bind labile standard proteins. Complex formation does not interfere with immunogenic detection and, importantly, antigenic as well as enzymatic activity remains constant for weeks. This strategy seems to be applicable to a wide range of assays involving unstable proteins, including the generation of vaccines. |
| Databáze: | OpenAIRE |
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