ES2525945T3 - Purificación cromatográfica de eritropoyetina humana recombinante - Google Patents

Purificación cromatográfica de eritropoyetina humana recombinante Download PDF

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Publication number
ES2525945T3
ES2525945T3 ES02803796.8T ES02803796T ES2525945T3 ES 2525945 T3 ES2525945 T3 ES 2525945T3 ES 02803796 T ES02803796 T ES 02803796T ES 2525945 T3 ES2525945 T3 ES 2525945T3
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ES
Spain
Prior art keywords
epohr
column
fractions
eluting
supernatant
Prior art date
Legal status (The legal status 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 status listed.)
Expired - Lifetime
Application number
ES02803796.8T
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English (en)
Inventor
Peter Alliger
Norbert Palma
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sandoz AG
Original Assignee
Sandoz AG
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 Sandoz AG filed Critical Sandoz AG
Application granted granted Critical
Publication of ES2525945T3 publication Critical patent/ES2525945T3/es
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

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Classifications

    • 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
    • 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
    • C07K1/20—Partition-, reverse-phase or hydrophobic interaction chromatography
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/505—Erythropoietin [EPO]
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

Abstract

Método para recuperar y purificar eritropoyetina humana recombinante (EPOhr) a partir de un medio de cultivo celular que comprende células huésped, método que comprende las etapas de: (a) eliminar células huésped, constituyentes celulares y residuos del medio de cultivo celular realizando un procedimiento seleccionado del grupo que consiste en (i) centrifugación seguida por una etapa de filtración en profundidad, (ii) una etapa de filtración en profundidad y (iii) centrifugación, para obtener un sobrenadante de medio de cultivo clarificado; (b) ajustar la conductividad del sobrenadante a 5 mS/cm o menos, y un pH de entre aproximadamente 7,0 y 8,0; (c) aplicar el sobrenadante de la etapa (b) a una columna que comprende un medio cromatográfico de intercambio aniónico, lavar la columna, eluir la EPOhr de la columna y recoger la fracción/fracciones de picos que contienen EPOhr; (d) someter las fracciones de picos combinadas de la etapa (c) a una etapa de cromatografía de fase inversa usando una resina que puede ejecutarse a presión media (< 10 bar) y es resistente a altas concentraciones de NaOH, eluyéndose la EPOhr usando un gradiente lineal de un disolvente orgánico; (e) aplicar una o más fracciones eluidas en la etapa (d) que contienen EPOhr a una columna que comprende medios cromatográficos de intercambio aniónico, lavar la columna y eluir la EPOhr usando un gradiente salino lineal; (f) seleccionar una o más fracciones eluidas en la etapa (e) que contienen EPOhr basándose en el grado de sialilación de la EPOhr; y (g) someter una o más fracciones eluidas en la etapa (f) que contienen EPOhr a una o más etapas cromatográficas de exclusión molecular usando un medio de filtración en gel para eliminar posibles dímeros y agregados superiores; y recoger el eluato que contiene EPOhr

Description

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E02803796
19-12-2014
(continuación)
Transfección
Número de clones por T25 Qp, MTX 9,6x10 -8 M [µg/106 células/d]
09/T25/3 (5:1)
459 0,14
09/T25/4 (5:1)
648 0,9
Transfecciones en 96 pocillos
Transfección
Número de clones por placa Clon seleccionado
09/96/1 (50:1)
47 1F5
09/96/2 (50:1)
46
09/96/3 (50:1)
50 5D5, 3H5
09/96/4 (50:1)
52
09/96/5 (50:1)
49 5D4, 5H4
09/96/6 (5:1)
416 6C5
09/96/7 (5:1)
556 7E9
09/96/8 (5:1)
392
09/96/9 (5:1)
427
09/96/10 (5:1)
352
09/96/11 (75:1)
49
09/96/12 (75:1)
60 12A9
Amplificación de diferentes clones
Clon
Qp [µg/10 células/d]
MTX 9,6x10 -8 M
MTX 1,9x10 -7 M MTX 3,8x10 -7 M
09/96/1F5
11 11,4 17,1
09/96/3D5
8,4 14,4 11,4
09/96/3H5
8,2 14,1 12,9
09/96/5D4
4,9 3,8 3,6
09/96/5H1
4,7 7,1 6,7
09/96/6C5
4,2 3,8 3,6
30
imagen29

Claims (1)

  1. imagen1
    imagen2
ES02803796.8T 2001-11-28 2002-11-26 Purificación cromatográfica de eritropoyetina humana recombinante Expired - Lifetime ES2525945T3 (es)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US33383901P 2001-11-28 2001-11-28
US333839P 2001-11-28
PCT/EP2002/013299 WO2003045996A1 (en) 2001-11-28 2002-11-26 Chromatographic purification of recombinant human erythropoietin

Publications (1)

Publication Number Publication Date
ES2525945T3 true ES2525945T3 (es) 2015-01-02

Family

ID=23304479

Family Applications (1)

Application Number Title Priority Date Filing Date
ES02803796.8T Expired - Lifetime ES2525945T3 (es) 2001-11-28 2002-11-26 Purificación cromatográfica de eritropoyetina humana recombinante

Country Status (20)

Country Link
US (1) US20060099674A1 (es)
EP (1) EP1453857B1 (es)
JP (1) JP4293908B2 (es)
KR (1) KR20040065567A (es)
CN (1) CN1596268A (es)
AU (1) AU2002356733A1 (es)
BR (1) BR0214568A (es)
CA (1) CA2466627A1 (es)
ES (1) ES2525945T3 (es)
HR (1) HRP20040476A2 (es)
HU (1) HUP0500061A2 (es)
IL (1) IL161855A0 (es)
MX (1) MXPA04005189A (es)
NO (1) NO20042185L (es)
NZ (1) NZ533083A (es)
PL (1) PL368755A1 (es)
RU (1) RU2004119818A (es)
SI (1) SI1453857T1 (es)
WO (1) WO2003045996A1 (es)
ZA (1) ZA200403461B (es)

Families Citing this family (27)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4109204B2 (ja) * 2002-03-26 2008-07-02 レツク・フアーマシユーテイカルズ・デー・デー 所望エリスロポエチングリコアイソフォームプロフィールの製造方法
DE60228460D1 (de) * 2002-12-13 2008-10-02 Bioceuticals Arzneimittel Ag Verfahren zur Herstellung und Reinigung von Erythropoietin
US7157276B2 (en) * 2003-06-20 2007-01-02 Biogen Idec Inc. Use of depth filtration in series with continuous centrifugation to clarify mammalian cell cultures
KR100567448B1 (ko) * 2003-11-05 2006-04-04 주식회사 인투젠 형질전환 동물의 유즙 전처리방법 및 이를 통한 인체단백질의 정제방법
DE102004027816A1 (de) 2004-06-08 2006-01-05 Bioceuticals Arzneimittel Ag Verfahren zur Reinigung von Erythropoietin
JP4742042B2 (ja) * 2004-06-30 2011-08-10 テルモ株式会社 ウィルス不活化ヘモグロビンおよびその製造方法
US7531632B2 (en) * 2006-02-16 2009-05-12 Gtc Biotherapeutics, Inc. Clarification of transgenic milk using depth filtration
KR100988706B1 (ko) * 2007-03-07 2010-10-18 한미홀딩스 주식회사 재조합 에리트로포이에틴의 신규한 정제방법
KR100900033B1 (ko) * 2007-11-21 2009-06-01 한국생명공학연구원 인간 에리스로포이에틴의 정제방법
DE102008002209A1 (de) 2008-06-04 2009-12-10 Evonik Degussa Gmbh Verfahren zur Aufreinigung von Erythropoietin
RU2011102447A (ru) 2008-06-24 2012-07-27 Др.Реддис' Лабораторис Лтд. (In) Очистка модифицированных цитокинов
EP2337787A1 (en) * 2008-10-17 2011-06-29 Percivia LLC. Clarification process
DE102008054716A1 (de) * 2008-12-16 2010-06-17 Evonik Degussa Gmbh Inprozesskontrolle in einem Verfahren zur Herstellung von EPO
US20120264688A1 (en) * 2009-09-23 2012-10-18 Walter Hinderer Process for the purification of recombinant human erythropoietin (epo), epo thus purified and pharmaceutical compositions comprising same
WO2011063195A2 (en) * 2009-11-20 2011-05-26 Dr. Reddy's Laboratories Ltd. Purification of modified cytokines
CN102453087B (zh) * 2010-10-22 2013-12-25 广东赛保尔生物医药技术有限公司 一种单取代peg-epo的纯化及制备方法
WO2012068134A1 (en) * 2010-11-15 2012-05-24 Biogen Idec Inc. Enrichment and concentration of select product isoforms by overloaded bind and elute chromatography
WO2012069474A2 (en) * 2010-11-24 2012-05-31 H. Lundbeck A/S A method for reducing potential virus burden in a sample by cyanate treatment
KR101443257B1 (ko) 2011-10-18 2014-09-19 주식회사 종근당 낮은 등전점을 갖는 에리스로포이에틴 유사체의 정제방법
WO2014138921A1 (en) * 2013-03-15 2014-09-18 Apotex Inc. Enhanced liquid formulation stability of erythropoietin alpha through purification processing
MX2016000063A (es) 2013-07-05 2016-03-01 Lab Francais Du Fractionnement Matriz de cromatografia de afinidad.
KR101414897B1 (ko) 2013-11-29 2014-07-04 씨제이헬스케어 주식회사 다베포에틴 알파의 정제 방법
WO2015134640A1 (en) * 2014-03-05 2015-09-11 GlycoScientific LLC Isotopically labeled glycans and methods for producing the same
CA2970732C (en) 2014-12-15 2023-05-16 Merck Patent Gmbh Target molecule capture from crude solutions
EP3153522A1 (en) 2015-10-09 2017-04-12 Hetero Drugs Limited Process for the purification of erythropoietin and darbepoetin alfa
CN106770597A (zh) * 2017-03-01 2017-05-31 上海博威生物医药有限公司 用于检测组合物中促红细胞生成素电荷异质体的cze检测方法
CN112940101A (zh) * 2019-12-11 2021-06-11 深圳赛保尔生物药业有限公司 重组人促红细胞生成素的纯化方法

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4677195A (en) * 1985-01-11 1987-06-30 Genetics Institute, Inc. Method for the purification of erythropoietin and erythropoietin compositions
US5856298A (en) * 1989-10-13 1999-01-05 Amgen Inc. Erythropoietin isoforms
EP0443724B1 (en) * 1990-02-20 1999-03-17 Baxter International Inc. Viral-safe purified human thrombin
DE19504211A1 (de) * 1995-02-09 1996-08-14 Behringwerke Ag Entfernen von Viren durch Ultrafiltration aus Proteinlösungen
TW442492B (en) 1995-07-26 2001-06-23 Novartis Ag Process for the enrichment of interferon
DK0889949T3 (da) * 1996-03-13 2003-09-22 Delta Biotechnology Ltd Fermenteringskontrol
BR9917606A (pt) 1998-11-06 2002-12-31 Bio Sidus S A Procedimento para a purificação de eritropoetina humana recombinante a partir de sobrenadantes de cultivo de células e eritropoetina humana recombinante obtida com tal procedimento

Also Published As

Publication number Publication date
ZA200403461B (en) 2006-05-31
HUP0500061A2 (hu) 2005-04-28
RU2004119818A (ru) 2006-01-10
HRP20040476A2 (en) 2005-10-31
PL368755A1 (en) 2005-04-04
JP2005518196A (ja) 2005-06-23
BR0214568A (pt) 2004-11-03
EP1453857A1 (en) 2004-09-08
WO2003045996A1 (en) 2003-06-05
CA2466627A1 (en) 2003-06-05
NZ533083A (en) 2006-09-29
SI1453857T1 (sl) 2015-01-30
IL161855A0 (en) 2005-11-20
NO20042185L (no) 2004-08-23
EP1453857B1 (en) 2014-08-13
MXPA04005189A (es) 2005-02-17
JP4293908B2 (ja) 2009-07-08
KR20040065567A (ko) 2004-07-22
CN1596268A (zh) 2005-03-16
US20060099674A1 (en) 2006-05-11
AU2002356733A1 (en) 2003-06-10

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