DK2332996T3 - Oprensning af anti-Her2-antistoffer - Google Patents

Oprensning af anti-Her2-antistoffer Download PDF

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Publication number
DK2332996T3
DK2332996T3 DK10187798.3T DK10187798T DK2332996T3 DK 2332996 T3 DK2332996 T3 DK 2332996T3 DK 10187798 T DK10187798 T DK 10187798T DK 2332996 T3 DK2332996 T3 DK 2332996T3
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Denmark
Prior art keywords
buffer
polypeptide
antibody
wash
salt concentration
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DK10187798.3T
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English (en)
Inventor
Jefferson C Emery
Paul J Mcdonald
Rhona O'leary
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Genentech Inc
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First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=31994112&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=DK2332996(T3) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
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Publication of DK2332996T3 publication Critical patent/DK2332996T3/da

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C07K1/16—Extraction; Separation; Purification by chromatography
    • C07K1/18—Ion-exchange chromatography
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
    • B01D15/36—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving ionic interaction
    • B01D15/361—Ion-exchange
    • B01D15/362—Cation-exchange
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
    • B01D15/36—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving ionic interaction
    • B01D15/361—Ion-exchange
    • B01D15/363—Anion-exchange
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oncology (AREA)
  • Cell Biology (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Claims (18)

1. Fremgangsmåde til oprensning af et polypeptid fra en sammensætning, omfattende polypeptidet og forurenende stoffer, hvor polypeptidet er et antistof, som binder HER2, og hvor de forurenende stoffer omfatter en deamide-ret variant af antistoffet, og hvor fremgangsmåden omfatter følgende fortløbende trin: (a) lade sammensætningen på en kationsbytningsharpiks med en ækvilibre-ringsbuffer, som har en første saltkoncentration; (b) vaske kationsbytningsharpiksen med en vaskebuffer, indtil en forudbestemt protein koncentration er målt i gennemstrømningen, hvor saltkoncentrationen af vaskebufferen stiger fra en indledende, anden saltkoncentration, som er større end saltkoncentrationen af ækvilibreringsbufferen, til en endelig, tredje saltkoncentration; (c) lede en fikseret volumen fra 0,4 til 1 søjlevoluminer afvaskebufferen med den endelige, tredje saltkoncentration gennem kationsbytningsharpiksen; og (d) eluere polypeptidet fra kationsbytningsharpiksen med en elueringsbuffer, som har en saltkoncentration, som er større end den endelige saltkoncentration af vaskebufferen.
2. Fremgangsmåde ifølge krav 1, hvorved kationsbytningsharpiksen omfatter carboxy-methylcellulose eller sulphopropyl immobiliseret på agarose.
3. Fremgangsmåde ifølge krav 1 eller krav 2, hvorved antistoffet omfatter en letkæde-aminosyresekvens SEQ ID NO: 1 og en tungkæde-aminosyresekvens SEQ ID NO: 2.
4. Fremgangsmåde ifølge et af kravene 1 til 3, hvorved antistoffet er rhuMAb HER2 omfattende en letkæde-aminosyresekvens SEQ ID NO: 1 og en tungkæde-aminosyresekvens SEQ ID NO: 2, og hvor de forurenende stoffer omfatter en deamideret variant, som har Asn30 i CDR1 af den ene eller begge af VL-regionerne deraf konverteret til aspartat.
5. Fremgangsmåde ifølge et af de foregående krav, hvorved antistoffet produceres rekombinant i CHO-værtsceller.
6. Fremgangsmåde ifølge krav 5, hvorved antistoffet produceres intracellu-lært og adskilles fra partikelformede rester af værtscellerne ved centrifugering eller ultrafiltrering inden ionbytningskromatografi.
7. Fremgangsmåde ifølge et af de foregående krav, hvorved antistoffet udsættes for protein A kromatografi inden kationsbytningskromatografi.
8. Fremgangsmåde ifølge et af de foregående krav, hvorved mængden af antistof i sammensætningen, der lades på ionbytningsharpiksen, er fra 15 mg til 45 mg per ml af kationbytningsharpiksen.
9. Fremgangsmåde ifølge et af de foregående krav, hvorved den forudbestemte protein koncentration i trin (b) svarer til en OD på 0,6 målt ved 280 nm.
10. Fremgangsmåde ifølge et af de foregående krav, hvorved vaskebufferen omfatter en blanding af en ækvilibreringsbuffer og elueringsbuffer, og hvorved stigningen afvaskebufferens saltkoncentration i trin (b) opnås ved at øge andelen af elueringsbufferen i vaskebufferen.
11. Fremgangsmåde ifølge krav 10, hvorved andelen af elueringsbufferen i vaskebufferen stiger med en konstant rate.
12. Fremgangsmåde ifølge krav 10, hvorved procentsatsen af elueringsbufferen i vaskebufferen stiger med to eller flere forskellige rater under vaskningen i trin (b).
13. Fremgangsmåde ifølge krav 12, hvorved procentsatsen af elueringsbufferen i vaskebufferen stiger med en første rate for et første segment af vaskningen, med en anden rate for et andet segment af vaskningen og med en tredje rate for et tredje segment af vaskningen.
14. Fremgangsmåde ifølge et af de foregående krav, hvorved elueringsbufferen omfatter 145 mM Na/FIOAc, og ækvilibreringsbufferen omfatter 70 mM Na/FIOAc, eller hvorved elueringsbufferen omfatter 100 mM NaCI, og ækvilibreringsbufferen omfatter 45 mM NaCI.
15. Fremgangsmåde ifølge et af de foregående krav, hvilken fremgangsmåde desuden omfatter det at udsætte sammensætningen, omfattende polypepti-det, for et eller flere oprensningstrin, således at der opnås en homogen fremstilling af polypeptidet.
16. Fremgangsmåde ifølge krav 15, hvilken fremgangsmåde desuden omfatter fremstilling af en farmaceutisk sammensætning ved at kombinere den homogene fremstilling af polypeptidet med en farmaceutisk acceptabel bærer.
17. Fremgangsmåde ifølge krav 15, hvilken fremgangsmåde desuden omfatter konjugering af det oprensede polypeptid med et heterologt molekyle.
18. Fremgangsmåde ifølge krav 17, hvorved det heterologe molekyle er po-lyethylenglycol, en label eller et cytotoksisk middel.
DK10187798.3T 2002-09-11 2003-09-08 Oprensning af anti-Her2-antistoffer DK2332996T3 (da)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US41033402P 2002-09-11 2002-09-11
EP03795664.6A EP1543038B2 (en) 2002-09-11 2003-09-08 Protein purification

Publications (1)

Publication Number Publication Date
DK2332996T3 true DK2332996T3 (da) 2014-12-15

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DK10187798.3T DK2332996T3 (da) 2002-09-11 2003-09-08 Oprensning af anti-Her2-antistoffer
DK03795664.6T DK1543038T4 (da) 2002-09-11 2003-09-08 Proteinoprensning

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Country Status (15)

Country Link
US (10) US8044017B2 (da)
EP (3) EP2332996B1 (da)
JP (7) JP5303092B2 (da)
AU (2) AU2003265994B2 (da)
CA (1) CA2496060C (da)
CY (1) CY1119024T1 (da)
DK (2) DK2332996T3 (da)
ES (2) ES2527616T3 (da)
HK (1) HK1158223A1 (da)
HU (1) HUE033623T2 (da)
IL (5) IL262513B (da)
LT (1) LT1543038T (da)
PT (1) PT1543038T (da)
SI (1) SI1543038T2 (da)
WO (1) WO2004024866A2 (da)

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