Laboratory of biotechnology of glycoproteins

Department of biotechnology

Head: Vorob'ev I.I.

recombinant proteins, eukaryotic producers, coagulation factors, hormones

The main research area of ​​the laboratory is the development of methodological approaches to obtaining Chinese hamster CHO ovary cell line for production pharmaceutically significant glycoproteins in large quantities.

The methods developed in the laboratory made it possible to obtain a number of cell lines secreting human coagulation factors VIII and IX, several gonadotropic heterodimeric hormones, and a number of other glycosylated proteins. In the case of follicle-stimulating human hormone (FSH), the producer line obtained by the laboratory was used in the full-scale industrial production of the bio-analog drug FSH "Primapur".

An original series of plasmid vectors has been developed that allows efficient integration into the eukaryotic genome and amplification of the target protein genes in it, as well as providing an extremely high level of expression of the target polypeptide through the use of the non-coding DNA of the Chinese hamster housekeeping gene and a concatemer fragment of the long terminal repeat of the Eppstein-Barr virus . The properties of these vectors were studied using model fluorescent proteins, which allowed them to be used in practical developments in the most rational way.

Before molecular cloning, the genes of the target proteins are preliminarily analyzed to select a cDNA variant capable of providing the maximum level of transcription and translation (selection of effective codons for producer cells, determination of the optimal number of introns). In the obtained producer cells, copying, translation and transcription efficiency of the target gene are determined. The resulting producer lines are examined for the selection of optimal cultivation conditions and stability. In addition, the laboratory is developing methods for purification of target proteins from the culture medium, including pharmacopoeial indicators.

Basic research methods
As part of the research, the laboratory implements the full cycle of the development of biotechnological methods - from obtaining highly effective producers of biologically active substances to developing methods for the isolation of target proteins. This cycle includes both traditional and special (including those developed in the laboratory) methods and approaches.
 

Research Methods:
1) analysis, optimization and generation of target protein genes;
2) molecular cloning, methods of working with NA (Real time-PCR, southern blot, etc.);
3) obtaining, culturing and researching the efficiency and stability of producer lines, selection optimal conditions for the expression of target proteins;
4) microscopic methods for the study of producer lines using fluorescent proteins;
5) chromatographic methods for purification target proteins;
6) immunochemical methods for the analysis of the efficiency of producers and final preparations (ELISA, western blot);
7) chromatographic methods for determining the content of impurities in protein preparations
A Brief History of the Laboratory
The laboratory was established in 2019 in the field of biotechnology. The head of the laboratory is Doctor of Biological Sciences Vorobyov I.I.

1. Obtaining highly effective producers of biologically active protein nature substances

2. Development of methods for purification target proteins

3. Obtaining modified preparations of prolonged action proteins

In the laboratory are developed:

 

Highly effective producers of coagulation factors VIII and IX for the treatment of hemophilia.
A producer of human follicle-stimulating hormone (FSH), which is widely used in medicine for the treatment of infertility and with the use of assisted reproductive technologies, significantly superior in terms of productivity to known analogues.
The producers of luteinizing human hormone and thyroid-stimulating hormone.
Producer of a full-sized human plasminogen tissue activator.
Producer of a full-sized human plasminogen tissue activator.
It is known that modifications such as pegylation and sialylation can increase the lifetime of the protein in the blood of patients. Therefore, the laboratory has developed a method for producing sialylated insulin, which can significantly increase the periods between drug injections, which improves the quality of patient's life . A method for the preparation of a sialylated derivative of the peptide hormone oxyntomodulin was also previously developed, the synthesized conjugate showed a multiple increase in the period of anorexic action compared with unmodified oxyntomodulin.

 

INNOVATIVE DEVELOPMENTS

Status

Development's Name

Date

Introduction's place 

Short description

1

Implemented (The drug is registered, put on the market)

The drug follicle-stimulating human hormone (FSH) Primapur® (follitropin alpha) 2019

LLC "iVFarma", Moscow

Recombinant human follicle-stimulating hormone is used as a stimulator of ovules formation in the body of a woman in in vitro fertilization (IVF) programs. The drug is obtained in the cells of a Chinese hamster. In the laboratory, a line of FSH producer was developed; methods for the isolation, purification and quality control of a protein preparation were developed.

2 Planned for implementation

The drug

of tissue activator of human plasminogen (alteplase)
2017 SPA"Microgen", Moscow

Producer of human plasminogen tissue activator and method for its purification.

 

RESULTS
Registration number

Type

 

Title

 

Authors

Applicant / Patent Holder

 

Priority date

Publication Date

 

1 2585532 RF patent Plasmid for expression of recombinant human coagulation factor IX, monoclonal cell line of a mammal - producer of recombinant human coagulation factor IX and method for producing said factor

Kovnir S.V., Orlova N.A., Vorobiev I.I., Khodak Y.A., Dronina M. A., Smirnov I. V., Ponomarenko N. A., Skryabin K.G.,  Gabibov A. G. 

FRC Bioengineering RAS, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS 31.01.2014

27.05.2016

2 2560596 RF patent Plasmid for expression recombinant human follicle-stimulating hormone (FSH) hormone in CHO cells, plasmid for expression of recombinant human FSH beta-subunit in CHO cells, CHO cell - producer of recombinant human FSH and method for producing said hormone

Vorobiev I.I., Orlova N.A., Kovnir S.V., Khodak Y.A.

LLC "iVFarma", Moscow

24.09.2013

27.03.2015

 

FullnamePositionContacts
Vorob'ev I.I., D.ScHead of lab.
Muzyka I.S.r. f.
Lyutsiya Dayanovaeng.
Loading...
Loading...

Vorob'ev I.I.

Russia, Moscow, Ul. Miklukho-Maklaya 16/10 — On the map

Loading...