Subject: research project

From: "Nekrasov Alex" <[email-masked]>
Date: Wed, 21 Feb 1996 18:11:25

Dear Collegues:

	We would like to collaborate with you and submit a joint research 
proposal to U.S. Civilian Research and Development Foundation 
(WWW: http://www.internext.com/crdf).
We look forward to beginning the dialogue and then to expanding our research 
and business relations. If you are interested in cooperation in these mutual 
investigation we will greatly appreciate you kindly reply by FAX or E-mail. 
Please do not reply to the mailing list because we are not subscribed.

Dr. A. Wulfson, Ph.D.
Head of the Recombinant Proteins Group
FAX:  +7 (095) 330-7274
E-mail:  [email-masked]
_____________________________________________________________________________
General:  our research group of biotechnology of recombinant proteins deals 
with isolation, purification and production of biologically active human 
recombinant proteins such as cytokines and insulin. Recently we carried out 
the development of a technological scheme for the production of human 
TNF-alpha (wild-type as well as  mutants), Il-3 and IFN-gamma. Based upon 
strains designed at our Institute, we are developing at present the 
technological operation for production of insulin. Results of our 
investigation of TNF-alpha will be published in 
Russian Journal of Bioorganic  Chemistry N3, 1996 
(English edition is available)
"An Effective Method of Purification For Human Recombinant Tumor Necrosis 
Factor Alpha".
R.V. Tikhonov, S.A. Yakimov, V.G. Korobko, A.N. Wulfson
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, 
Russian Academy of Sciences, 117871, Miklukho-Maklaya St. 16/10, 
Moscow, Russia.

During the progress of our research work we have faced some problems 
connected with preservation of biological activity of  rhIFN-gamma. 
That's only one reason why we have no opportunity to start the clinical 
trials of our product and make promotion of the medicines based on rhIFN-gamma 
for clinical applications. To solve this problem we are going to make 
experimental work to design more stable variants of  rhIFN-gamma in the 
following lines of investigation:

1. COMPUTER DESIGN OF PROTEINS
The computer modeling by the molecular dynamics method of protein spatial 
structure to determine potential mutation regions which allow formation of 
rhIFN-gamma dimer binding by S-S bonds.

2. GENETIC ENGINEERING
a) Preparation of  N-terminal deletion mutants lacking 7-10 residues. We 
suppose that such shortening will increase the stability of protein and 
facilitate its renaturation without lost of protein activity 
b). Preparation of mutant analogs of rhIFN-gamma in which the spatial 
structure would be stabilized by intermolecular S-S bonds.

3. STRUCTURAL ANALYSIS
Analysis  of  the protein dimer  spatial structure by  X-ray analysis  or  
NMR-spectroscopy.

4. BIOTECHNOLOGY
Isolation and purification of the recombinant protein, its renaturation and 
determination of reagents and buffer solutions in which the protein remains 
stable during long time periods and can be lyophilized with minimal lost of 
activity.

We hope that a combination of these approaches would give us the possibility 
to design stable substance of protein rhIFN-gamma for our further work.