A Purified, Biologically Active Peg Gcsf And Processes Thereof

Abstract: ABSTRACT: The present invention provides an improved process for expressing, isolating and purifying PEG-G-CSF from a G-CSF producing microorganism. The invention also relates to a method of expression of G-CSF in E. coli, including sequence, plasmids, expression vectors and host cells for use in such an improved method. The invention also relates to methods used to clone gene encoding the target protein; inserting the related codon-optimsed gene in a suitable E.coli host; producing culture of the recombinant strain, stimulating expression of the protein, its secretion and purification post fermentation and related processes such as conjugation with PEG. More particularly, the invention relates to a simplified and optimized purification process with respect to solubilization of inclusion bodies and refolding procedures and subsequent chromatographic purification methods, the described chromatographic procedures enabling high yields, selectivity and purity of the desired end product.

Patent Information

Application #

Filing Date

30 March 2009

Publication Number

42/2010

Publication Type

INA

Invention Field

MICRO BIOLOGY

Status

Email

Parent Application

Applicants

BIOCON LIMITED

20th KM Hosur Road Electronic City P.O. Bangalore – 560 100 Karnataka India

Inventors

1. SUDHIR MANJUL

Flat No. 204 Mahaveer Regency 4th A Main 6th Cross Bilekahalli Bangalore -560076 Karnataka India

2. ANUPAMA REDDY

D/O D M Srinivasa Reddy # 41 "Matru Nilaya" Shabarinagar II Cross Byatarayanapura Bellary Road Bangalore – 560092 Karntaka India.

3. NAGARAJ GOVINDAPPA

Kodagikere Durvigere Post Channagiri Tq. Davanagere Dist. Karnataka State PIN: 577213

4. PARTHA HAZRA

Apartment Number S-19 Meenakshi Residency 41/1 Arekere Bannerghata Road Bangalore 560076 Karnataka India.

5. HARISH IYER

5C-221 3rd Main Road HRBR 3rd Block Bangalore 560043 Karnataka India.

Specification

FIELD OF THE INVENTION:
The present invention provides an improved process for expressing, isolating and purifying PEG-G-CSF from a G-CSF producing microorganism. The invention also relates to a method of expression of G-CSF in E. coli, including sequence, plasmids, expression vectors and host cells for use in such an improved method. The invention also relates to methods used to clone gene encoding the target protein; inserting the related codon-optimsed gene in a suitable E.coli host; producing culture of the recombinant strain, stimulating expression of the protein, its secretion and purification post fermentation and related processes such as conjugation with PEG. More particularly, the invention relates to a simplified and optimized purification process with respect to solubilization of inclusion bodies and refolding procedures and subsequent chromatographic purification methods, the described chromatographic procedures enabling high yields, selectivity and purity of the desired end product.

Documents

Application Documents

#	Name	Date
1	731-CHE-2009_EXAMREPORT.pdf	2016-07-02
1	Form-5.pdf	2011-09-03
2	731-CHE-2009 CORRESPONDENCE OTHERS 28-05-2014.pdf	2014-05-28
2	Form-3.pdf	2011-09-03
3	Form-1.pdf	2011-09-03
3	731-CHE-2009 CORRESPONDENCE OTHERS 28-05-2014.pdf	2014-05-28
4	Drawings.pdf	2011-09-03
4	731-CHE-2009 FORM-1 28-05-2014.pdf	2014-05-28
5	731-CHE-2009 FORM-13 13-07-2012.pdf	2012-07-13
5	731-CHE-2009 FORM-13 28-05-2014.pdf	2014-05-28
6	731-CHE-2009 CORRESPONDENCE OTHERS 19-02-2014.pdf	2014-02-19
6	731-CHE-2009 FORM-1 13-07-2012.pdf	2012-07-13
7	731-CHE-2009 POWER OF ATTORNEY 19-02-2014.pdf	2014-02-19
7	731-CHE-2009 CORRESPONDENCE OTHERS 13-07-2012.pdf	2012-07-13
8	731-CHE-2009 AMENDED CLAIMS 28-01-2014.pdf	2014-01-28
8	731-CHE-2009 CORRESPONDENCE OTHERS 28-01-2014.pdf	2014-01-28
9	731-CHE-2009 AMENDED PAGES OF SPECIFICATION 28-01-2014.pdf	2014-01-28
10	731-CHE-2009 CORRESPONDENCE OTHERS 28-01-2014.pdf	2014-01-28
10	731-CHE-2009 AMENDED CLAIMS 28-01-2014.pdf	2014-01-28
11	731-CHE-2009 POWER OF ATTORNEY 19-02-2014.pdf	2014-02-19
11	731-CHE-2009 CORRESPONDENCE OTHERS 13-07-2012.pdf	2012-07-13
12	731-CHE-2009 CORRESPONDENCE OTHERS 19-02-2014.pdf	2014-02-19
12	731-CHE-2009 FORM-1 13-07-2012.pdf	2012-07-13
13	731-CHE-2009 FORM-13 13-07-2012.pdf	2012-07-13
13	731-CHE-2009 FORM-13 28-05-2014.pdf	2014-05-28
14	Drawings.pdf	2011-09-03
14	731-CHE-2009 FORM-1 28-05-2014.pdf	2014-05-28
15	Form-1.pdf	2011-09-03
15	731-CHE-2009 CORRESPONDENCE OTHERS 28-05-2014.pdf	2014-05-28
16	Form-3.pdf	2011-09-03
16	731-CHE-2009 CORRESPONDENCE OTHERS 28-05-2014.pdf	2014-05-28
17	Form-5.pdf	2011-09-03
17	731-CHE-2009_EXAMREPORT.pdf	2016-07-02