Novo Proteins with Interferon
Design, Genetic Engineering, Production and Physico-Chemical Testing of Biologically Active de Novo Proteins Incorporating an Alfa-Interferon Fragment. Biological Study of the Obtained Constructions
Tech Area / Field
- BIO-CGM/Cytology, Genetics and Molecular Biology/Biotechnology
- MED-DRG/Drug Discovery/Medicine
8 Project completed
Senior Project Manager
Institute of Immunological Engineering, Russia, Moscow reg., Lyubuchany
- Institute of Bioorganic Chemistry, Russia, Moscow
- Leiras Oy / Biomedical Center, Finland, Turku\nGesellschaft für Biotechnologische Forschung mbH, Germany, Braunschweig\nUniversity of Medicine and Dentistry of New Jersey (UMD) / Robert Wood Johnson Medical School, USA, NJ, Piscataway\nLabmaster Ltd., Finland, Turku\nAmersham International, UK, Bucks, Little Chalfont
Project summaryThe design of de novo proteins with predetermined structure and properties is one of the most intriguing challenges of modern protein engineering. At present, a number of such proteins have been designed, and a few of them have been produced and studied (for a review see C.Sander, 1994, Trends in Biotechnologyt 12, 163-167). The next step in this direction should be the production of a protein possessing not only the predetermined structure but a biological activity as well. Such constructions could possess useful biological and pharmacologic properties as candidates for new-type potential medicines. Two years ago we have engineered the first biologically active construction based on a de novo protein albebetin with predesigned structure and an octapeptide fragment of human a2-interferon which activates blast transformation of thymocyte cells. We have obtained nanogram amounts of the radiolabeled chimeric protein in a cell-free translational system. Its initial study using a new approach developed by us showed that the protein possesses compact and relatively stable structure and biological activity even higher than that of intertferon at a concentration as low as 10-11 M (D.A.Dolgikh et al., 1993, Biofizika (Russian) 38, 67-74) . These data demonstrate a necessity of a detailed investigation of biological, physical and chemical properties of the constructed biologically active protein. To this end we need first of all an efficient expression system to produce the protein in preparative scale (10-50 mg). On the other hand we plan to design and produce the modified biologically active constructions based on the results of our study and molecular modeling in order to improve the structural and biological properties of the proteins.
Objective of the project:
Production in preparative scale and study of biologically active protein constructions, including a-peptoferon and the de novo protein albebetin.
The results expected:
- production of preparative amounts of biologically active de novo proteins based on de novo protein albebetin and a-peptoferon, for biological studies and preclinical trials.
- production of biologically active de novo proteins with stable, proteolysis-resistant structure with valuable pharmacological properties. A de novo protein carrying the only grafted biological activity of interest has no side effects unlike natural proteins possessing usually a number of various activities. Besides that, de novo proteins designed from the basic principles as carriers of biological activities would possess lower immunogenity than natural proteins.