KR960010682A - 금속이온 친화성 크로마토그라피를 이용한 히루딘의 정제방법 - Google Patents

금속이온 친화성 크로마토그라피를 이용한 히루딘의 정제방법 Download PDF

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Publication number
KR960010682A
KR960010682A KR1019940022499A KR19940022499A KR960010682A KR 960010682 A KR960010682 A KR 960010682A KR 1019940022499 A KR1019940022499 A KR 1019940022499A KR 19940022499 A KR19940022499 A KR 19940022499A KR 960010682 A KR960010682 A KR 960010682A
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KR
South Korea
Prior art keywords
hirudin
purifying
phosphate buffer
affinity chromatography
metal ion
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Application number
KR1019940022499A
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English (en)
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KR0141651B1 (ko
Inventor
이상기
정봉현
최의성
손정훈
윤덕중
김명국
이해돈
Original Assignee
강재헌
동국제약 주식회사
김은영
한국과학기술연구원
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.)
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Publication date
Application filed by 강재헌, 동국제약 주식회사, 김은영, 한국과학기술연구원 filed Critical 강재헌
Priority to KR1019940022499A priority Critical patent/KR0141651B1/ko
Priority to DE19532495A priority patent/DE19532495A1/de
Priority to JP7228173A priority patent/JPH08176199A/ja
Priority to FR9510394A priority patent/FR2724175A1/fr
Priority to US08/524,311 priority patent/US5698104A/en
Priority to GB9518225A priority patent/GB2292942B/en
Priority to CN95117701A priority patent/CN1071337C/zh
Publication of KR960010682A publication Critical patent/KR960010682A/ko
Application granted granted Critical
Publication of KR0141651B1 publication Critical patent/KR0141651B1/ko

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/81—Protease inhibitors
    • C07K14/815—Protease inhibitors from leeches, e.g. hirudin, eglin
    • 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
    • C07K1/22—Affinity chromatography or related techniques based upon selective absorption processes
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
    • Y10S530/855—Proteins from animals other than mammals or birds

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Analytical Chemistry (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

본 발명은 히루딘을 히루딘을 정제하는 방법을 제공한다. 본 발명의 방법은 조(粗) 히루딘액을 금속 이온으로서 구리이온을 사용하고 용리액으로서 인산염 완충액을 사용하는 금속이온 친화성 크로마토그래피를 이용하여 정제함을 특징으로 한다. 본 발명의 방법에 의하면 종래의 히루딘의 정제방법에 비해 저렴하고 간단하게 히루딘을 고순도로 정제할 수 있는 장점이 있다.

Description

금속이온 친화성 크로마토그라피를 이용한 히루딘의 정제방법
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음

Claims (5)

  1. 조(粗) 하루딘액을 금속 이온으로서 구리이온을 사용하고 용리액으로서 인산염 완충액을 사용하는 금속이온 친화성 크로마토그래피를 이용하여 정제함을 특징으로 하는 히루딘의 정제방법.
  2. 제1항에 있어서, 히루딘은 천연형, HV1,HV2 또는 Lys47-HV2히루딘임을 특징으로 하는 정제방법.
  3. 제1항에 있어서, 인산염 완충액은 농도가 100mM 미만인 인산염 완충액임을 특징으로 하는 정제방법.
  4. 리간드가 충전된 컬럼에 구리이온 용액을 흘려주어 구리이온을 리간드에 결합시키고; 조 히루딘 용액을 상기 컬럼에 주입하여 히루딘을 구리이온에 결합시키며; 인산염 완충액을 이용하여 히루딘을 구리이온으로부터 탈착시킴을 특징으로 하는 히루딘의 정제방법.
  5. 제4항에 있어서, 리간드는 이미노디아세트산임을 특징으로 하는 정제방법.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019940022499A 1994-09-07 1994-09-07 금속이온 친화성 크로마토그라피를 이용한 히루딘의 정제방법 KR0141651B1 (ko)

Priority Applications (7)

Application Number Priority Date Filing Date Title
KR1019940022499A KR0141651B1 (ko) 1994-09-07 1994-09-07 금속이온 친화성 크로마토그라피를 이용한 히루딘의 정제방법
DE19532495A DE19532495A1 (de) 1994-09-07 1995-09-02 Verfahren zum Reinigen von Hirudin
JP7228173A JPH08176199A (ja) 1994-09-07 1995-09-05 アフィニティークロマトグラフィーを用いたヒルジンの精製法
FR9510394A FR2724175A1 (fr) 1994-09-07 1995-09-05 Procede de purification de l'hirudine par chromatographie d'affinite
US08/524,311 US5698104A (en) 1994-09-07 1995-09-06 Purification process for hirudin using affinity chromatography
GB9518225A GB2292942B (en) 1994-09-07 1995-09-06 Purification process for hirudin using affinity chromatography
CN95117701A CN1071337C (zh) 1994-09-07 1995-09-07 用亲合色谱法提纯水蛭素的方法

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
KR1019940022499A KR0141651B1 (ko) 1994-09-07 1994-09-07 금속이온 친화성 크로마토그라피를 이용한 히루딘의 정제방법

Publications (2)

Publication Number Publication Date
KR960010682A true KR960010682A (ko) 1996-04-20
KR0141651B1 KR0141651B1 (ko) 1998-06-15

Family

ID=19392218

Family Applications (1)

Application Number Title Priority Date Filing Date
KR1019940022499A KR0141651B1 (ko) 1994-09-07 1994-09-07 금속이온 친화성 크로마토그라피를 이용한 히루딘의 정제방법

Country Status (7)

Country Link
US (1) US5698104A (ko)
JP (1) JPH08176199A (ko)
KR (1) KR0141651B1 (ko)
CN (1) CN1071337C (ko)
DE (1) DE19532495A1 (ko)
FR (1) FR2724175A1 (ko)
GB (1) GB2292942B (ko)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6479300B1 (en) * 1999-03-15 2002-11-12 Millipore Corporation Metal loaded ligand bound membranes for metal ion affinity chromatography
WO2006045503A1 (en) 2004-10-19 2006-05-04 Lonza Ag Method for solid phase peptide synthesis
CN100405060C (zh) * 2006-01-24 2008-07-23 李振国 一种水蛭指纹图谱的建立方法和水蛭药材的鉴别方法
CN102153645B (zh) * 2010-12-24 2013-03-06 南宁乙翔生物科技有限公司 用电渗析除去水蛭素中重金属离子的方法

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA1231306A (en) * 1983-03-03 1988-01-12 Erich Hochuli Purification of interferon
DE3738541A1 (de) * 1987-11-13 1989-05-24 Hoechst Ag Verfahren zur isolierung und reinigung von hirudin
US4832849A (en) * 1988-06-16 1989-05-23 Merrell Dow Pharmaceuticals Inc. Extraction and purification of an anticoagulant principle from the south american leech, haementeria ghilianii
US5169936A (en) * 1989-04-14 1992-12-08 Biogen, Inc. Protein purification on immobilized metal affinity resins effected by elution using a weak ligand
US5179196A (en) * 1989-05-04 1993-01-12 Sri International Purification of proteins employing ctap-iii fusions
US5077388A (en) * 1990-05-31 1991-12-31 Schering Corporation Purification of human interleukin-4 from a secreted escherichia coli mutant
DE4001238A1 (de) * 1990-01-18 1991-07-25 Basf Ag Verfahren zur anreicherung bzw. reinigung von biomolekuelen
US5034133A (en) * 1990-07-26 1991-07-23 Schering-Corporation Purification of human interleukin-4 from a CHO-cell line culture medium
US5372719A (en) * 1992-03-30 1994-12-13 Perseptive Biosystems, Inc. Molecular imaging

Also Published As

Publication number Publication date
CN1071337C (zh) 2001-09-19
US5698104A (en) 1997-12-16
DE19532495A1 (de) 1996-03-14
FR2724175A1 (fr) 1996-03-08
CN1127260A (zh) 1996-07-24
JPH08176199A (ja) 1996-07-09
GB2292942B (en) 1998-09-02
KR0141651B1 (ko) 1998-06-15
GB2292942A (en) 1996-03-13
GB9518225D0 (en) 1995-11-08

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