JPH06510671A - キメラ抗体の製造−組合せアプローチ - Google Patents
キメラ抗体の製造−組合せアプローチInfo
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- C12N15/09—Recombinant DNA-technology
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- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1037—Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
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Abstract
Description
Claims (23)
- 1.着目の抗原に特異的な抗体ポリペプチド二量体の製造方法であって、次の段 階: (i)複製可能な遺伝表示パッケージ(rgdp)の成分を使ってパッケージン グすることができる核酸発現ベクターを提供し;(ii)(a)各々ヒト抗体の 第一の抗原結合部位構成部分をコードする核酸配列の遺伝学上多様なレパートリ ーを、(b)前記着目の抗原を結合することが知られている非ヒト抗体の第二の 抗原結合部位構成部分のユニーク集団または遺伝学上多様な集団をコードする核 酸と組み合わせ、前記発現ベクター上に抗体ポリペプチド二量体をコードする核 酸配列のライブラリーを形成せしめ、ここで前記二量体は各々第一のポリペプチ ド鎖成分と第二のポリペプチド鎖成分とから成り、前記第一の抗原結合部位構成 部分と前記第二の抗原結合部位構成部分は一緒になって抗原ポリペプチド二量体 の抗原結合部位を形成し; (iii)組換え宿主生物細胞において前記ベクターから前記ライブラリーを発 現せしめ、前記第一のポリペプチド鎖成分の各々が、前記第一のポリペプチド鎖 成分をrdgpの表面に表示するrgdpの成分との融合体として発現され; (iv)前記発現されたライブラリーから、前記着目の抗原に対する結合特異性 を有する前記抗体ポリペプチド二量体のユニーク集団または限定集団を抗原との 結合により選択し、各選択された抗体ポリペプチド二量体が各々のrgdp中で それの前記第一の抗原結合部位構成部分をコードする核酸と関連づけられるを含 んで成る方法。
- 2.前記抗体ポリペプチド二量体が段階(iv)での選択後に可溶性ポリペプチ ドとして発現される、請求項1に記載の方法。
- 3.前記段階(ii)での組合せの前に、各々前記非ヒト抗体の第二の構成成分 をコードする配列を遺伝子的に変更してヒト抗体の第二構成部分との相同性を増 加させる、請求項1または請求項2に記載の方法。
- 4.前記段階(iv)において選択された抗体ポリペプチド二量体を遺伝子的に 変更して非ヒト特性を除去または削減する追加の段階(v)を有する、請求項1 〜3のいずれか一項に記載の方法。
- 5.前記段階(v)が、 (a)段階(iv)において選択された前記第一の抗原結合部位構成部分をコー ドする核酸のユニーク集団または限定集団を、各々ヒト抗体の第二の抗原結合部 位構成部分をコードする核酸配列の遺伝学上多様なレパートリーと組合せ、抗体 ポリペプチド二量体をコードする核酸配列の第二のライブラリーを形成せしめ、 ここで各抗体ポリペプチドニ量体は、rgdpの一成分を使ってパッケージング することができる核酸配列から発現される第二の抗体ポリペプチド鎖成分を含ん で成り、前記第二の鎖成分は、それによってrgdpの表面に前記第二の鎖成分 を表示するrgdpの成分との融合体として発現され、その結果、前記着目の抗 原との結合によって前記第二のライブラリーから着目の前記抗原に特異的な抗体 ポリペプチド二量体を選択することが可能であり、各抗体ポリペプチドニ量体が そのrgdp中でそれの第二のポリペプチド成分をコードする核酸と関連づけら れることを含んで成る、請求項4に記載の方法。
- 6.前記非ヒト抗体の第二の抗原結合部位構成成分が抗原との接触を行う領域で ある、請求項1〜5のいずれか一項に記載の方法。
- 7.前記領域が相補性決定領域(CDR)である、請求項6に記載の方法。
- 8.前記非ヒト抗体の第二の抗原結合部位構成成分が一抗体の第二のポリペプチ ド鎖成分である、請求項1〜5のいずれか一項に記載の方法。
- 9.前記抗体ポリペプチド二量体のライブラリーのいずれかの各抗体ポリペプチ ド二量体が一本のポリペプチド鎖、好ましくはscFv断片として発現される、 請求項1〜8のいずれか一項に記載の方法。
- 10.前記抗体ポリペプチド二量体のライブラリーのいずれかの各抗体ポリペプ チド二量体が二本のポリペプチド鎖、好ましくはFvまたはFab断片として発 現される、請求項1〜8のいずれか一項に記載の方法。
- 11.着目の抗原に特異的なヒト抗体ポリペプチド二量体の製造方法であって、 (i)各々ヒト抗体の第一のポリペプチド鎖の可変ドメインを含んで成るポリペ プチドの多様性集団(集団A)を、各々前記抗原に特異的な非ヒト抗体の第二の ポリペプチド鎖の可変ドメインを含んで成るポリペプチドのユニーク集団または 限定集団(集団B)と組合せ、それによって各々ヒト抗体の第一のポリペプチド 鎖の可変ドメインを含んで成るポリペプチドと非ヒト抗体の第二のポリペプチド 鎖の可変ドメインを含んで成るポリペプチドとから成る抗体ポリペプチド二量体 のライブラリーを形成せしめ:(ii)前記ライブラリーから、前記抗原に対す る結合特異性を有する前記抗体ポリペプチド二量体のユニーク集団または限定集 団(集団C)を選択し; (iii)各々ヒトの第一のポリペプチド鎖を含んで成る段階(ii)において 選択されたポリペプチド二量体から誘導したポリペプチドのユニーク集団または 限定集団(集団D)を、各々ヒト抗体の第二のポリペプチド鎖の可変ドメインを 含んで成るポリペプチドの多様性集団(集団E)と組合せ、それによってヒト抗 体ポリペプチド鎖二量体のライブラリーを形成せしめ、そのライブラリーから、 前記抗原に特異的なヒト抗体ポリペプチド二量体のユニーク集団または限定集団 (集団F)を選択することが可能であることを含んで成る方法。
- 12.(a)前記集団Aのポリペプチドが、組換え宿主生物細胞において、それ によってrgdpの第一集団においてrgdpの表面上に前記ポリペプチドを表 示する複製可能な遺伝表示パッケージ(rgdp)の成分との融合体としてベク ター(X)から発現され、(b)前記rgdp成分を使って前記ベクター(X) の核酸をパッケージングすることができ、それによって前記rgdpの遺伝物質 が前記集団Aのポリペプチドをコードし、その結果、集団Cの抗体ポリペプチド 二量体がそれら各々のrgdp中でヒト抗体の第一のポリペプチド鎖の可変領域 を含んで成るポリペプチドをコードする核酸と関連づけられ; (c)前記集団Eのポリペプチドの各々が、組換え宿主生物細胞において、rg dpの第二集団においてrgdpの表面上に前記ポリペプチドを表示するrgd pの成分との融合体としてベクター(Y)から発現され;そして (d)前記rgdp成分を使って前記ベクター(Y)の核酸をパッケージングす ることができ、それによって、rgdpの第二集団の中の前記rgdpの遺伝物 質が前記集団Eのポリペプチドをコードする請求項11に記載の方法。
- 13.前記集団Aが前記集団Bと同じベクターから発現されず;そして前記集団 Dが前記集団Eと同じベクターから発現されない、請求項11または請求項12 に記載の方法。
- 14.前記集団Aが前記集団Bと同じベクターから発現され;そして前記集団D が前記集団Eと同じベクターから発現される、請求項11または請求項12に記 載の方法。
- 15.前記抗体ポリペプチド二量体のライブラリーのいずれかの各抗体ポリペプ チド二量体が一本のポリペプチド鎖、好ましくはscFv断片として発現される 、請求項14に記載の方法。
- 16.前記抗体ポリペプチド二量体のライブラリーのいずれかの各抗体ポリペプ チド二量体が二本のポリペプチド鎖、好ましくはFvまたはFab断片として発 現される、請求項11〜14のいずれか一項に記載の方法。
- 17.前記集団Bのポリペプチドが各々ヒト抗体定常ドメインを含んで成るキメ ラである、請求項11または請求項12に記載の方法。
- 18.前記集団Eのポリペプチドが、各々前記抗原に特異的な非ヒト抗体からの 領域を含んで成り、前記領域が抗原との接触を行う領域である、請求項11また は請求項12に記載の方法。
- 19.前記領域が相補性決定領域(CDR)である、請求項18に記載の方法。
- 20.前記CDRがCDR3である、請求項19に記載の方法。
- 21.前記第一のポリペプチド鎖が抗体軽鎖であり、前記第二のポリペプチド鎖 が抗体重鎖である、請求項11〜20のいずれか一項に記載の方法。
- 22.前記抗原に特異的なヒト抗体ポリペプチド二量体の前記集団Fを選択する 追加の段階を含んで成る、請求項11〜21のいずれか一項に記載の方法。
- 23.前記集団CとFのいずれか一方が前記抗原との結合により選択される、請 求項11〜22のいずれか一項に記載の方法。
Applications Claiming Priority (11)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB919120252A GB9120252D0 (en) | 1991-09-23 | 1991-09-23 | Improvement binding substances |
GB9120252.3 | 1991-09-23 | ||
GB919120377A GB9120377D0 (en) | 1991-09-25 | 1991-09-25 | Improved binding substances |
GB9120377.8 | 1991-09-25 | ||
GB9206318.9 | 1992-03-24 | ||
GB929206372A GB9206372D0 (en) | 1992-03-24 | 1992-03-24 | Binding substances |
GB929206318A GB9206318D0 (en) | 1992-03-24 | 1992-03-24 | Binding substances |
PCT/GB1992/000883 WO1992020791A1 (en) | 1990-07-10 | 1992-05-15 | Methods for producing members of specific binding pairs |
GB92/00883 | 1992-05-15 | ||
PCT/GB1992/001755 WO1993006213A1 (en) | 1991-09-23 | 1992-09-23 | Production of chimeric antibodies - a combinatorial approach |
GB9206372.6 | 1992-09-23 |
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JPH06510671A true JPH06510671A (ja) | 1994-12-01 |
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JP50593293A Expired - Lifetime JP3540315B2 (ja) | 1991-09-23 | 1992-09-23 | キメラ抗体の製造−組合せアプローチ |
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JP (1) | JP3540315B2 (ja) |
AT (1) | ATE181571T1 (ja) |
AU (1) | AU665025B2 (ja) |
CA (1) | CA2119930C (ja) |
DE (1) | DE69229477T2 (ja) |
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JP2008512382A (ja) * | 2004-09-03 | 2008-04-24 | ジェネンテック・インコーポレーテッド | ヒト化抗β7アンタゴニストおよびその使用 |
WO2014054820A1 (ja) * | 2012-10-03 | 2014-04-10 | 株式会社リブテック | in vivoで抗腫瘍活性を有する抗ヒトDlk-1抗体 |
US9303086B2 (en) | 2012-10-03 | 2016-04-05 | Livtech, Inc. | Anti-hDlk-1 antibody having an antitumor activity in vivo |
JPWO2014054820A1 (ja) * | 2012-10-03 | 2016-08-25 | 株式会社カイオム・バイオサイエンス | invivoで抗腫瘍活性を有する抗ヒトDlk−1抗体 |
KR20150060761A (ko) * | 2012-10-03 | 2015-06-03 | 가부시키가이샤 리부텍쿠 | in vivo 에서 항종양 활성을 갖는 항인간 Dlk-1 항체 |
Also Published As
Publication number | Publication date |
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AU2593392A (en) | 1993-04-27 |
AU665025B2 (en) | 1995-12-14 |
ATE181571T1 (de) | 1999-07-15 |
WO1993006213A1 (en) | 1993-04-01 |
DE69229477T2 (de) | 1999-12-09 |
CA2119930A1 (en) | 1993-04-01 |
CA2119930C (en) | 2002-10-01 |
DK0605522T3 (da) | 2000-01-17 |
EP0605522B1 (en) | 1999-06-23 |
JP3540315B2 (ja) | 2004-07-07 |
EP0605522A1 (en) | 1994-07-13 |
DE69229477D1 (de) | 1999-07-29 |
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