ME02162B - Proces prečišćavanja proteina - Google Patents

Proces prečišćavanja proteina

Info

Publication number
ME02162B
ME02162B MEP-2015-94A MEP9415A ME02162B ME 02162 B ME02162 B ME 02162B ME P9415 A MEP9415 A ME P9415A ME 02162 B ME02162 B ME 02162B
Authority
ME
Montenegro
Prior art keywords
host cell
recombinant
recombinant protein
seq
extract
Prior art date
Application number
MEP-2015-94A
Other languages
English (en)
Inventor
Gavin Barry Wild
Original Assignee
Ucb Biopharma Sprl
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Ucb Biopharma Sprl filed Critical Ucb Biopharma Sprl
Publication of ME02162B publication Critical patent/ME02162B/me

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Classifications

    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K1/00General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14Extraction; Separation; Purification
    • C07K1/30Extraction; Separation; Purification by precipitation
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K1/00General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/107General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K1/00General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14Extraction; Separation; Purification
    • C07K1/16Extraction; Separation; Purification by chromatography
    • C07K1/18Ion-exchange chromatography
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K1/00General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14Extraction; Separation; Purification
    • C07K1/16Extraction; Separation; Purification by chromatography
    • C07K1/22Affinity chromatography or related techniques based upon selective absorption processes
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K1/00General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14Extraction; Separation; Purification
    • C07K1/36Extraction; Separation; Purification by a combination of two or more processes of different types
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475Growth factors; Growth regulators
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K16/00Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K16/00Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/24Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
    • C07K16/241Tumor Necrosis Factors
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/90Isomerases (5.)
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00Preparation of peptides or proteins
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2317/00Immunoglobulins specific features
    • C07K2317/10Immunoglobulins specific features characterized by their source of isolation or production
    • C07K2317/14Specific host cells or culture conditions, e.g. components, pH or temperature
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2317/00Immunoglobulins specific features
    • C07K2317/50Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/55Fab or Fab'

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Immunology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Peptides Or Proteins (AREA)

Claims (25)

1. Metod za prečišćavanje rekombinantnog proteina iz gram-negativnog bakterijskog uzorka ćelije domaćina ili njegovog ekstrakta pri čemu navedena ćelija domaćina eksprimira rekombinantni protein i rekombinantnu disulfid izomerazu DsbC; pri čemu metoda sadrži: a) podešavanje pH uzorka ćelije domaćina, ili njegovog ekstrakta do pH 5 ili manje da se istaloži rekombinantna disulfid izomeraza; i b) odvajanje istaložene rekombinantne disulfid izomeraze iz rekombinantnog proteina da se proizvede uzorak rekombinantnog proteina.
2. Metod prema patentnom zahtevu 1, pri čemu je pH uzorka ćelije domaćina ili njegovog ekstrakta podešena na pH od 4.5 ili manje.
3. Metod prema patentnom zahtevu 2, pri čemu je pH uzorka ćelije domaćina ili njegovog ekstrakta podešena na pH od 4.5 do 3.0.
4. Metod prema patentnom zahtevu 2, pri čemu je pH uzorka ćelije domaćina ili njegovog ekstrakta podešena na pH od 3 ili manje.
5. Metod prema patentnom zahtevu 4, pri čemu se u koraku a) dipeptid ćelije domaćina vezujućeg proteina je dalje istaložen i ukoraku b) dipeptid ćelije domaćina je dalje odvojen od rekombinantnog proteina.
6. Metod prema bilo kojem od prethodnih patentnih zahteva, pri čemu disulfid izomeraza sadrži dodatak histidina.
7. Metod prema bilo kojem od prethodnih patentnih zahteva, pri čemu je gram-negativna bakterijska ćelija domaćina E. koli.
8. Metod prema bilo kojem od prethodnih patentnih zahteva, pri čemu ćelija domaćina sadrži Fkpa i/ili Skp.
9. Metod prema bilo kojem od prethodnih patentnih zahteva, pri čemu u koraku a) pH uzorka ćelije domaćina ili njegov ekstrakt je podešen na pH od manje od 5 upotrebom glacijalne-ledene sirćetne kiseline.
10. Metod prema bilo kojem od prethodnih patentnih zahteva, pri čemu uzorak ćelije domaćina ili njegov ekstrakt se drži na pH od 5 ili manje tokom jednog sata ili manje.
11. Metod prema bilo kojem od prethodnih patentnih zahteva, pri čemu separacija u koraku b) sadrži centrifugiranje i/ili filtriranje.
12. Metod prema bilo kojem od prethodnih patentnih zahteva, pri čemu nakon koraka b) metod sadrži dalji korak podvrgavanja prečišćavanju uzorka rekombinantnog proteina do hromatografije.
13. Metod prema patentnom zahtevu 11, pri čemu je hromatografija hromatografija jonske izmene.
14. Metod prema bilo kojem od prethodnih patentnih zahteva, pri čemu nakon koraka b) pH uzorka rekombinantnog proteina je podešena na pH od 5 do 7.
15. Metod prema bilo kojem od prethodnih patentnih zahteva, pri čemu pre koraka a) pufer za ekstrakciju je dodat uzorku ćelije domaćina ili njegovom ekstraktu i uzorak ćelije domaćina ili njegov ekstrakt je podvrgnut koraku termičke obrade.
16. Metod prema bilo kojem od prethodnih patentnih zahteva, pri čemu rekombinantni protein ima pl od 6 do 9.
17. Metod prema patentnom zahtevu 16, pri čemu rekombinantni protein ima pl od 8 do 9.
18. Metod prema bilo kojem od prethodnih patentnih zahteva, pri čemu rekombinantni protein je rekombinantno antitelo.
19. Metod prema patentnom zahtevu 18, pri čemu je rekombinantno antitelo monoklonalno, humanizovano ili himerno antitelo, ili fragment od njih, kao što je vezujući fragment od njih.
20. Metod prema patentnom zahtevu 18 ili zahtevu 19, pri čemu je rekombinantno antitelo Fab ili Fab-ov fragment.
21. Metod prema bilo kojem od patentnih zahteva 18 do 20, pri čemu je rekombinantno antitelo specifično za humani TNFɑ.
22. Metod prema patentnom zahtevu 21, pri čemu antitelo sadrži teški lanac gde varijabilni domen sadrži CDR koji ima sekvencu datu u SEQ ID NO:1 za CDRH1, SEQ ID NO:2, za CDRH2 i SEQ ID NO:3 za CDRH3 i laki lanac pri čemu varijabilni domen sadrži CDR koji ima sekvencu datu u SEQ ID NO:4 za CDRL1, SEQ ID NO:5 za CDRL2 i SEQ ID NO:6 za CDRL3.
23. Metod prema patentnom zahtevu 22, pri čemu antitelo sadrži sekvencu varijabilne regije lakog lanca datu u SEQ ID NO: 7 i varijabilna regija teškog lanca je data u SEQ ID NO: 8.
24. Metod prema patentnom zahtevu 23, pri čemu je antitelo Fab fragment i sadrži teški lanac koji ima sekvencu datu u SEQ ID NO: 10 i laki lanac koji ima sekvencu datu u SEQ ID NO: 9.
25. Upotreba koraka podešavanja pH od gram-negativnog bakterijskog uzorka ćelije domaćina ili njegovog ekstrakta transformisanog ekspresonim vektorom koji kodira rekombinantni protein na pH 5 ili manje da se istaloži rekombinantna disulfid izomeraza ćelije domaćina, odvoji precipitirana rekombinantna disulfid izomeraza ćelije domaćina iz rekombinantnog proteina i proizvede prečišćeni uzorak rekombinantnog proteina.
MEP-2015-94A 2010-07-27 2011-07-27 Proces prečišćavanja proteina ME02162B (me)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
GBGB1012599.5A GB201012599D0 (en) 2010-07-27 2010-07-27 Process for purifying proteins
PCT/GB2011/001129 WO2012013930A2 (en) 2010-07-27 2011-07-27 Process for purifying proteins
EP11745810.9A EP2598516B8 (en) 2010-07-27 2011-07-27 Process for purifying proteins

Publications (1)

Publication Number Publication Date
ME02162B true ME02162B (me) 2015-10-20

Family

ID=42752868

Family Applications (1)

Application Number Title Priority Date Filing Date
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Country Status (24)

Country Link
US (1) US9751930B2 (me)
EP (1) EP2598516B8 (me)
JP (1) JP5935222B2 (me)
KR (1) KR101944121B1 (me)
CN (1) CN103189385B (me)
AU (1) AU2011284548B2 (me)
BR (1) BR112013000177B1 (me)
CA (1) CA2804099C (me)
CY (1) CY1116526T1 (me)
DK (1) DK2598516T3 (me)
ES (1) ES2537877T3 (me)
GB (1) GB201012599D0 (me)
HR (1) HRP20150639T1 (me)
HU (1) HUE026049T2 (me)
IL (1) IL224044A (me)
ME (1) ME02162B (me)
MX (1) MX337184B (me)
PL (1) PL2598516T3 (me)
PT (1) PT2598516E (me)
RS (1) RS54085B1 (me)
SG (2) SG186974A1 (me)
SI (1) SI2598516T1 (me)
SM (1) SMT201500142B (me)
WO (1) WO2012013930A2 (me)

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Publication number Publication date
US9751930B2 (en) 2017-09-05
RS54085B1 (sr) 2015-10-30
GB201012599D0 (en) 2010-09-08
MX2013000899A (es) 2013-05-22
SG186974A1 (en) 2013-02-28
KR20130103713A (ko) 2013-09-24
IL224044A (en) 2016-09-29
WO2012013930A3 (en) 2012-05-03
CA2804099A1 (en) 2012-02-02
ES2537877T3 (es) 2015-06-15
PT2598516E (pt) 2015-07-22
AU2011284548B2 (en) 2015-11-26
HRP20150639T1 (hr) 2015-07-17
US20130178607A1 (en) 2013-07-11
SMT201500142B (it) 2015-09-07
CN103189385A (zh) 2013-07-03
SI2598516T1 (sl) 2015-07-31
EP2598516A2 (en) 2013-06-05
AU2011284548A1 (en) 2013-02-07
EP2598516B8 (en) 2015-07-15
EP2598516B1 (en) 2015-03-25
CN103189385B (zh) 2016-06-15
CA2804099C (en) 2018-07-10
JP5935222B2 (ja) 2016-06-15
JP2013535463A (ja) 2013-09-12
HUE026049T2 (en) 2016-05-30
SG196862A1 (en) 2014-02-13
DK2598516T3 (da) 2015-06-15
BR112013000177B1 (pt) 2021-11-09
WO2012013930A2 (en) 2012-02-02
KR101944121B1 (ko) 2019-01-30
PL2598516T3 (pl) 2015-10-30
CY1116526T1 (el) 2017-03-15
BR112013000177A2 (pt) 2020-09-24
MX337184B (es) 2016-02-16

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