JP2005170953A - L鎖欠落免疫グロブリン - Google Patents
L鎖欠落免疫グロブリン Download PDFInfo
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- JP2005170953A JP2005170953A JP2005010359A JP2005010359A JP2005170953A JP 2005170953 A JP2005170953 A JP 2005170953A JP 2005010359 A JP2005010359 A JP 2005010359A JP 2005010359 A JP2005010359 A JP 2005010359A JP 2005170953 A JP2005170953 A JP 2005170953A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- 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
- C07K16/20—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans from protozoa
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/22—Immunoglobulins specific features characterized by taxonomic origin from camelids, e.g. camel, llama or dromedary
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
-
- 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
- C07K2317/50—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
- C07K2317/567—Framework region [FR]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
- C07K2317/64—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising a combination of variable region and constant region components
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
-
- 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
- Y10S530/866—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof involving immunoglobulin or antibody fragment, e.g. fab', fab, fv, fc, heavy chain or light chain
Abstract
【解決手段】 天然に産する動物、ラクダの血清から精製され、天然4本鎖モデル免疫グロブリンと、もしくはその誘導体等の何れとも対応しない新規な免疫グロブリンを得ることが出来た。本免疫グロブリンは医学分野の診断、治療に使用し得る。
【選択図】 なし
Description
−発現ベクタープラスミドipBS(イムノPBS)(層細胞)は、プロモーターpLACによる制御下で、E. coli における免疫グロブリン鎖の発現に用いられるペル(pel)Bリーダー配列、リボゾーム結合部位及び終止コドンを含む。更に、C−末端デカペプチドタグの配列も含む。
−ipBS−VHH21プラスミドをもつE. coli JM101 を、アンピシリン100μg/ml及びグルコース0.1%を含むTB培地1l中32℃で発育させた。OD550 1.0でIPTG 1mM(最終濃度)の添加により、発現を誘発した。28℃で一晩誘発した後、4,000gで10分間(4℃)遠心分離することにより細胞を集め、TES緩衝液(pH8.0のトリス−HCl 0.2M、EDTA0.5mM、ショ糖0.5M)10ml中に再懸濁した。懸濁物を氷上に2時間保持した。周辺質タンパク質を、水により1:4容積/容積に希釈したTES緩衝液20mlの添加による浸透ショックにより除去し、1時間氷上に保持し、その後12,000g、4℃で30分間遠心分離した。上清の周辺質分画を、pH8.8のトリス−HCl、NaCl 50mMにより透析し、ファストQセファロースフロー(ファルマシア(Pharmacia))カラムに適用し、上述の緩衝液により洗浄し、緩衝液中で50mMから1M NaClのグラジエントにより溶離した。
Claims (31)
- 1以上の抗原を認識することができそして結合することができる2つの重ポリペプチド鎖を含むこと、及びその可変領域のアミノ酸配列が45位において荷電アミノ酸の中から選択されるか又はシステイン残基であるアミノ酸を含み、この免疫グロブリンが軽ポリペプチド鎖を欠くことを特徴とする、免疫グロブリン。
- ラクダ科のリンパ球又は他の細胞から得ることができるような軽鎖を欠く免疫グロブリンの配列をコードするDNA又はcDNAの原核生物又は真核生物宿主細胞における発現の産生物であることを特徴とする、請求項1に記載の免疫グロブリン。
- 各重鎖の各可変領域が少なくとも1の抗原結合性部位を含むことを特徴とする、請求項1又は2に記載の免疫グロブリン。
- ラクダ科の免疫グロブリンであることを特徴とする、請求項1〜3のいずれか1項に記載の免疫グロブリン。
- その定常領域の全部又は一部がヒト抗体の定常領域の全部又は一部によって置換されている、請求項1〜4のいずれか1項に記載の免疫グロブリン。
- 2つの重ポリペプチド鎖を含み、ポリペプチド鎖のそれぞれが異なる抗原を認識することができ、そして結合することができ、上記免疫グロブリンが軽ポリペプチド鎖を欠いていることを特徴とする、免疫グロブリン。
- 目的の抗原を結合し、そして正常軽鎖相互作用部位を欠く重ポリペプチド鎖の可変領域を含むことを特徴とする、免疫グロブリン。
- 抗原を認識することができ、そして結合することができる、請求項1〜4のいずれか1項に記載の免疫グロブリンの重鎖のポリペプチドに対応するフラグメント。
- 残基が荷電アミノ酸又はシステイン残基である上記重鎖の位置45に対応するアミノ酸残基を含む、請求項1〜7のいずれか1項に記載の免疫グロブリンの重鎖の可変領域の少なくとも10アミノ酸残基のフラグメント。
- 少なくとも20アミノ酸残基を含む、請求項9に記載のフラグメント。
- 抗原を認識することができ、そして結合することができる、免疫グロブリンの重鎖の可変領域である、請求項9又は10に記載のフラグメント。
- 免疫グロブリンのヒンジ領域又はこのヒンジ領域の少なくとも6アミノ酸に対応するフラグメントであるか、又はXがGluを除く任意のアミノ酸であるPro−Xの配列の反復配列を含むヒンジ領域フラグメントであることを特徴とする、請求項1〜7のいずれか1項に記載の免疫グロブリンのフラグメント。
- 請求項1〜7のいずれか1項に記載の免疫グロブリンの重ポリペプチド鎖の可変領域のフラグメントであって、CDR3ドメインを含み、そして上記重ポリペプチド鎖の位置45に対応するアミノ酸残基を含み、上記アミノ酸残基が、荷電アミノ酸及びシステイン残基からなる群から選択され、ここで、上記フラグメントが抗原に特異的な結合性部位を含む、フラグメント。
- 請求項1〜7のいずれか1項に記載の免疫グロブリンの可変領域(VHH)。
- 触媒活性を有することを特徴とする、請求項1〜14のいずれか1項に記載の免疫グロブリン又はフラグメント。
- 所定の基質の活性化状態を擬似する抗原に対して向けられたことを特徴とする、請求項15に記載の免疫グロブリン又はフラグメント。
- 細胞又は生命体において発現される、請求項1〜15のいずれか1項に記載の免疫グロブリン又はフラグメント。
- 酵母細胞、哺乳類細胞、昆虫細胞、原生動物細胞、植物細胞において得られる、請求項1〜15のいずれか1項に記載の免疫グロブリン又はフラグメント。
- 細菌、ウイルス、寄生生物などの抗原に対して向けられたか又はタンパク質、ハプテン、炭水化物又は核酸に対して向けられたことを特徴とする、請求項1〜18のいずれか1項に記載の免疫グロブリン又はフラグメント。
- 免疫グロブリンイディオタイプに対して向けられたことを特徴とする、請求項1〜18のいずれか1項に記載の免疫グロブリン又はフラグメント。
- 細胞性レセプター又は膜タンパク質に対して向けられたことを特徴とする、請求項1〜18のいずれか1項に記載の免疫グロブリン又はフラグメント。
- 毒素、酵素、薬物又はホルモンとコンジュゲーションしていることを特徴とする、請求項1〜18のいずれか1項に記載の免疫グロブリン又はフラグメント。
- 請求項1〜22のいずれか1項に記載の免疫グロブリン又はフラグメントを用いる、インビトロ診断のための方法。
- 請求項1〜22のいずれか1項に記載の免疫グロブリン又はそのフラグメントを投与する工程を含む、哺乳類におけるがんを処置するための方法であって、上記免疫グロブリン又はそのフラグメントが腫瘍特異的タンパク質に結合する、方法。
- 請求項1〜22のいずれか1項に記載の免疫グロブリン又はそのフラグメントを投与する工程を含み、上記免疫グロブリン又はそのフラグメントが病原性物質に結合する、哺乳類における病原性物質に対する防御を誘導するための方法。
- 請求項1〜22のいずれか1項に記載の免疫グロブリン又はフラグメントを投与する工程を含む、哺乳類におけるタンパク質の発現又は活性を調節するための方法。
- 請求項1〜22のいずれか1項に記載の免疫グロブリン又はそのフラグメントを投与する工程を含む、細胞の代謝を改変するための方法。
- 下記工程:
− 請求項1〜9のいずれか1項に記載の免疫グロブリンを産生することができ、決定された抗原で予め免疫したラクダ科のリンパ球から得られたDNA又はcDNA配列を、ベクター、特にファージ、そしてより詳細にはフィラメント状バクテリオファージ中へクローンニングする工程、
− 上記抗体の産生を可能にする条件で、原核細胞を上記ベクターで形質転換する工程、
− 形質転換細胞を抗原親和性選択に付すことによって適切な抗体を選択する工程、
− 所望の特異性を有する抗体を回収する工程
を含む、決定された抗原に対して向けられた免疫グロブリン又はフラグメントの調製のための方法。 - クローニングベクターがプラスミド又は真核生物ウイルスであり、形質転換細胞が真核生物細胞、特に酵母細胞、哺乳類細胞、植物細胞又は原生動物細胞である、請求項28に記載の方法。
- クローニングベクターが細菌膜において免疫グロブリンを発現することができるプラスミドである、請求項28に記載の方法。
- クローニングベクターが分泌タンパク質として免疫グロブリンを発現することができるプラスミドである、請求項28に記載の方法。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP92402326A EP0584421A1 (en) | 1992-08-21 | 1992-08-21 | Immunoglobulins devoid of light chains |
EP93401310 | 1993-05-21 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2001136457A Division JP3660270B2 (ja) | 1992-08-21 | 2001-05-07 | L鎖欠落免疫グロブリン |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2009192382A Division JP2009280608A (ja) | 1992-08-21 | 2009-08-21 | L鎖欠落免疫グロブリン |
Publications (3)
Publication Number | Publication Date |
---|---|
JP2005170953A true JP2005170953A (ja) | 2005-06-30 |
JP2005170953A5 JP2005170953A5 (ja) | 2005-11-04 |
JP4414900B2 JP4414900B2 (ja) | 2010-02-10 |
Family
ID=26132410
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP50590394A Expired - Fee Related JP3444885B2 (ja) | 1992-08-21 | 1993-08-18 | L鎖欠落免疫グロブリン |
JP2001136457A Expired - Lifetime JP3660270B2 (ja) | 1992-08-21 | 2001-05-07 | L鎖欠落免疫グロブリン |
JP2005010359A Expired - Lifetime JP4414900B2 (ja) | 1992-08-21 | 2005-01-18 | L鎖欠落免疫グロブリン |
JP2009192382A Pending JP2009280608A (ja) | 1992-08-21 | 2009-08-21 | L鎖欠落免疫グロブリン |
Family Applications Before (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP50590394A Expired - Fee Related JP3444885B2 (ja) | 1992-08-21 | 1993-08-18 | L鎖欠落免疫グロブリン |
JP2001136457A Expired - Lifetime JP3660270B2 (ja) | 1992-08-21 | 2001-05-07 | L鎖欠落免疫グロブリン |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2009192382A Pending JP2009280608A (ja) | 1992-08-21 | 2009-08-21 | L鎖欠落免疫グロブリン |
Country Status (13)
Country | Link |
---|---|
US (5) | US5800988A (ja) |
EP (6) | EP0656946B2 (ja) |
JP (4) | JP3444885B2 (ja) |
AT (5) | ATE452207T1 (ja) |
AU (1) | AU701578B2 (ja) |
CA (1) | CA2142331C (ja) |
DE (6) | DE69334258D1 (ja) |
DK (5) | DK0656946T4 (ja) |
ES (5) | ES2325541T3 (ja) |
FI (3) | FI115462B (ja) |
GR (1) | GR3037024T3 (ja) |
PT (5) | PT1498427E (ja) |
WO (1) | WO1994004678A1 (ja) |
Cited By (1)
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JP2009112204A (ja) * | 2007-11-02 | 2009-05-28 | Okayama Univ | 無機硫黄化合物加水分解酵素の製造方法 |
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EP0739981A1 (en) | 1995-04-25 | 1996-10-30 | Vrije Universiteit Brussel | Variable fragments of immunoglobulins - use for therapeutic or veterinary purposes |
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CA2429544C (en) * | 2000-11-17 | 2010-10-19 | University Of Rochester | In vitro methods of producing and identifying immunoglobulin molecules in eukaryotic cells |
US20060064782A1 (en) * | 2000-12-18 | 2006-03-23 | Conopco, Inc. | Production of antibodies |
AU2002235761A1 (en) | 2000-12-19 | 2002-07-01 | Unilever Plc | Stabilization of antibodies or fragments thereof |
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GB8607679D0 (en) * | 1986-03-27 | 1986-04-30 | Winter G P | Recombinant dna product |
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GB9016299D0 (en) * | 1990-07-25 | 1990-09-12 | Brien Caroline J O | Binding substances |
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JP2009112204A (ja) * | 2007-11-02 | 2009-05-28 | Okayama Univ | 無機硫黄化合物加水分解酵素の製造方法 |
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