JP2018138047A5 - - Google Patents
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- JP2018138047A5 JP2018138047A5 JP2018088350A JP2018088350A JP2018138047A5 JP 2018138047 A5 JP2018138047 A5 JP 2018138047A5 JP 2018088350 A JP2018088350 A JP 2018088350A JP 2018088350 A JP2018088350 A JP 2018088350A JP 2018138047 A5 JP2018138047 A5 JP 2018138047A5
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- 102100008989 IGHV1-2 Human genes 0.000 claims 20
- 101710003461 IGHV1-2 Proteins 0.000 claims 20
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 claims 20
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 claims 20
- 102000013463 Immunoglobulin Light Chains Human genes 0.000 claims 19
- 108010065825 Immunoglobulin Light Chains Proteins 0.000 claims 19
- 239000000427 antigen Substances 0.000 claims 16
- 102000038129 antigens Human genes 0.000 claims 16
- 108091007172 antigens Proteins 0.000 claims 16
- 230000000392 somatic Effects 0.000 claims 13
- 102000018358 Immunoglobulins Human genes 0.000 claims 11
- 108060003951 Immunoglobulins Proteins 0.000 claims 11
- 102000025417 antigen binding proteins Human genes 0.000 claims 11
- 108091000829 antigen binding proteins Proteins 0.000 claims 11
- 210000003719 B-Lymphocytes Anatomy 0.000 claims 10
- 102000004965 antibodies Human genes 0.000 claims 10
- 108090001123 antibodies Proteins 0.000 claims 10
- 210000004602 germ cell Anatomy 0.000 claims 10
- 210000004027 cells Anatomy 0.000 claims 9
- 230000003053 immunization Effects 0.000 claims 4
- 241000700159 Rattus Species 0.000 claims 2
- 238000004519 manufacturing process Methods 0.000 claims 2
Claims (26)
- 抗原結合性タンパク質を作製する方法であって、該方法は、
(a)遺伝子改変マウスのB細胞からヒト免疫グロブリン重鎖可変領域配列を取得するステップであって、
該遺伝子改変マウスは、
(i)該遺伝子改変マウスの生殖系列ゲノムにおいて単一の再構成されたヒトVκ1−39/Jκ配列と、
(ii)1つまたは複数のヒトV H 遺伝子セグメント、1つまたは複数のヒトD H 遺伝子セグメント、および1つまたは複数のヒトJ H 遺伝子セグメントと、
(iii)各々ヒト免疫グロブリン軽鎖可変ドメインを含む免疫グロブリン軽鎖を発現するB細胞の集団であって、該ヒト免疫グロブリン軽鎖可変ドメインは、該単一の再構成されたヒトVκ1−39/Jκ配列またはその体細胞超変異バリアントから発現され、該単一の再構成されたヒトVκ1−39/Jκ配列は、免疫グロブリン軽鎖定常領域配列に作動可能に連結されている、集団と
を含み、該B細胞は、該B細胞の集団の一部であり、該B細胞は、該ヒト免疫グロブリン重鎖可変領域配列を含む免疫グロブリン重鎖を発現し、該ヒト免疫グロブリン重鎖可変領域配列は、ヒトV H 2−26、V H 3−21、V H 3−64、またはその体細胞超変異バリアントを含む、ステップと、
(b)抗原結合性タンパク質の作製において(a)のヒト免疫グロブリン重鎖可変領域配列を使用するステップであって、該抗原結合性タンパク質は、
(i)該ヒト免疫グロブリン重鎖可変領域配列によってコードされるヒト免疫グロブリン重鎖可変ドメインを含む免疫グロブリン重鎖と、
(ii)該再構成されたヒトVκ1−39/Jκ配列またはその体細胞超変異バリアントによってコードされるヒト免疫グロブリン軽鎖可変ドメインを含む免疫グロブリン軽鎖と
を含む、ステップと
を含む方法。 - 前記集団の前記B細胞が、ヒト免疫グロブリン重鎖可変領域にコードされるヒト免疫グロブリン重鎖可変ドメインを含む免疫グロブリン重鎖を一緒に発現し、該ヒト免疫グロブリン重鎖可変領域は、V H 2−26またはその体細胞超変異バリアント、V H 3−21またはその体細胞超変異バリアント、およびV H 3−64またはその体細胞超変異を含む、請求項1に記載の方法。
- 前記マウスの生殖系列ゲノムにおける前記単一の再構成されたヒトVκ1−39/Jκ配列が、
(a)再構成されたヒトVκ1−39/Jκ5配列である;および/または
(b)マウスまたはラット免疫グロブリン軽鎖定常領域配列に作動可能に連結されている、
請求項1または請求項2に記載の方法。 - 前記単一の再構成されたヒトVκ1−39/Jκ配列が、
(a)内因性免疫グロブリンVκ遺伝子セグメントおよび内因性免疫グロブリンJκ遺伝子セグメントを置換するように配置されている;
(b)すべての機能的な内因性免疫グロブリンVκ遺伝子セグメントおよび内因性免疫グロブリンJκ遺伝子セグメントを置換するように配置されている;ならびに/あるいは
(c)マウスCκ領域に作動可能に連結している、
請求項1〜3のいずれか一項に記載の方法。 - 前記単一の再構成されたヒトVκ1−39/Jκ配列が、再構成されたヒトVκ1−39/Jκ5配列であり、該ヒトVκ1−39遺伝子セグメントと該ヒトJκ5遺伝子セグメントとの間のジャンクションにまたがる配列が、配列番号21に記載の配列を含む、請求項3に記載の方法。
- 前記マウスの生殖系列ゲノムは、前記単一の再構成されたヒトVκ1−39/Jκ配列についてホモ接合性である、請求項1〜5のいずれか一項に記載の方法。
- 前記ヒト免疫グロブリン重鎖可変領域が、ヒトV H 3−64またはその体細胞超変異バリアントを含む、請求項1〜6のいずれか一項に記載の方法。
- 前記ヒト重鎖可変領域が
(a)CH1、ヒンジ、CH2、CH3またはその組合せをコードするマウス免疫グロブリン重鎖定常(CH)領域配列に作動可能に連結している;ならびに/あるいは
(b)内因性免疫グロブリン重鎖遺伝子座に存在する、
請求項1〜7のいずれか一項に記載の方法。 - 前記遺伝子改変マウスの生殖系列ゲノムは、免疫グロブリン軽鎖の発現のために機能的ないかなる内因性免疫グロブリンVκ遺伝子セグメントを欠いており、前記遺伝子改変マウスの生殖系列ゲノムは、免疫グロブリン軽鎖の発現のために機能的ないかなる内因性免疫グロブリンJκ遺伝子セグメントを欠いている、請求項1〜8のいずれか一項に記載の方法。
- (a)遺伝子改変マウスのB細胞からヒト免疫グロブリン重鎖可変領域配列を取得する前に、前記遺伝子改変マウスを第1の目的の抗原で免疫化するステップをさらに含み、該B細胞が、該第1の目的の抗原に特異的に結合する第1の抗体を発現する、請求項1〜9のいずれか一項に記載の方法。
- 前記遺伝子改変マウスを第2の目的の抗原で免疫化するステップと、
第2のヒト免疫グロブリン重鎖可変領域配列が、前記集団の第2のB細胞から取得されるように、前記遺伝子改変マウスのB細胞からヒト免疫グロブリン重鎖可変領域配列を取得するステップであって、該第2のB細胞が、該第2の目的の抗原に特異的に結合する第2の抗体を発現する、ステップと、
抗原結合性タンパク質の作製において、該第2のヒト免疫グロブリン重鎖可変領域配列を使用するステップと
をさらに含む、請求項10に記載の方法。 - 前記第2のヒト免疫グロブリン重鎖可変領域配列が、ヒトV H 2−26、V H 3−21、V H 3−64、またはその体細胞超変異バリアントを含む、請求項11に記載の方法。
- 前記第1および第2の目的の抗原は、異なる抗原である、請求項11または12に記載の方法。
- 前記第1および第2の目的の抗原は、同じ抗原であり、前記第1および第2の抗体は、該同じ抗原の異なるエピトープに結合する、請求項11または12に記載の方法。
- 抗原結合性タンパク質を作製する方法であって、該方法は、
(a)第1の遺伝子改変マウスを第1の目的の抗原で免疫化するステップであって、該第1の遺伝子改変マウスは、B細胞の集団を含み、該集団のB細胞は、該第1の目的の抗原に特異的に結合する第1の抗体を発現し、
該第1の遺伝子改変マウスは、
(i)該第1の遺伝子改変マウスの生殖系列ゲノムにおいて単一の再構成されたヒトVκ1−39/Jκ配列と、
(ii)各々ヒト免疫グロブリン軽鎖可変ドメインを含む免疫グロブリン軽鎖を発現するB細胞の集団であって、該ヒト免疫グロブリン軽鎖可変ドメインは、該再構成されたヒトVκ1−39/Jκ配列またはその体細胞超変異バリアントから発現され、該再構成されたヒトVκ1−39/Jκ配列は、免疫グロブリン軽鎖定常領域配列に作動可能に連結されている、集団と
を含み、該第1の抗体を発現する該B細胞の集団のB細胞は、第1のヒト免疫グロブリン重鎖可変領域配列によってコードされる第1のヒト免疫グロブリン重鎖可変領域ドメインを含む第1の免疫グロブリン重鎖を発現し、該第1のヒト免疫グロブリン重鎖可変領域配列は、ヒトV H 2−26、V H 3−21、V H 3−64、またはその体細胞超変異バリアントを含む、ステップと、
(b)第2の遺伝子改変マウスを第2の目的の抗原で免疫化するステップであって、該第2の遺伝子改変マウスは、B細胞の集団を含み、該集団のB細胞は、該第2の目的の抗原に特異的に結合する第2の抗体を発現し、
該第2の遺伝子改変マウスは、
(i)該第2の遺伝子改変マウスの生殖系列ゲノムにおいて単一の再構成されたヒトVκ1−39/Jκ配列と、
(ii)各々ヒト免疫グロブリン軽鎖可変ドメインを含む免疫グロブリン軽鎖を発現するB細胞の集団であって、該ヒト免疫グロブリン軽鎖可変ドメインは、該再構成されたヒトVκ1−39/Jκ配列またはその体細胞超変異バリアントから発現され、該再構成されたヒトVκ1−39/Jκ配列は、免疫グロブリン軽鎖定常領域配列に作動可能に連結されている、集団と
を含み、該第2の抗体を発現する該B細胞の集団のB細胞は、第2のヒト免疫グロブリン重鎖可変領域配列によってコードされる第2の免疫グロブリン重鎖可変領域ドメインを含む第2の免疫グロブリン重鎖を発現する、ステップと、
(c)該第1の抗体由来の該第1のヒト免疫グロブリン重鎖可変ドメインを含む該第1の免疫グロブリン重鎖を選択し、かつ該第2の抗体由来の該第2のヒト免疫グロブリン重鎖可変ドメインを含む該第2の免疫グロブリン重鎖を選択するステップであって、該第1および第2のヒト免疫グロブリン重鎖可変ドメインは異なる、ステップと、
(d)該第1および第2のヒト免疫グロブリン重鎖可変ドメインの配列を決定するステップと、
(e)該第1および第2のヒト免疫グロブリン重鎖可変ドメインの配列を使用して、二重特異性抗原結合性タンパク質を作製するステップと
を含む方法。 - 前記第2のヒト免疫グロブリン重鎖可変領域配列は、ヒトV H 2−26、V H 3−21、V H 3−64、またはその体細胞超変異バリアントを含む、請求項15に記載の方法。
- 前記第1のマウスおよび前記第2のマウスは、同じマウスまたは異なるマウスである、請求項15に記載の方法。
- 前記第1および第2の目的の抗原は同じ抗原であり、前記第1および第2の抗体は、該同じ抗原の異なるエピトープに結合する、請求項15〜17のいずれか一項に記載の方法。
- 前記第1および第2の遺伝子改変マウスの生殖系列ゲノムにおける前記単一の再構成されたヒトVκ1−39/Jκ配列は、
(a)再構成されたヒトVκ1−39/Jκ5配列である;および/または
(b)マウスまたはラット免疫グロブリン軽鎖定常領域配列に作動可能に連結されている、
請求項15〜18のいずれか一項に記載の方法。 - 前記第1および第2の遺伝子改変マウスの前記単一の再構成されたヒトVκ1−39/Jκ配列は、
(a)内因性免疫グロブリンVκ遺伝子セグメントおよび内因性免疫グロブリンJκ遺伝子セグメントを置換するように配置されている;
(b)すべての機能的な内因性免疫グロブリンVκ遺伝子セグメントおよび内因性免疫グロブリンJκ遺伝子セグメントを置換するように配置されている;ならびに/あるいは
(c)マウスCκ領域に作動可能に連結している、
請求項15〜19のいずれか一項に記載の方法。 - 前記単一の再構成されたヒトVκ1−39/Jκ配列が、再構成されたヒトVκ1−39/Jκ5配列であり、該ヒトVκ1−39遺伝子セグメントと該ヒトJκ5遺伝子セグメントとの間のジャンクションにまたがる配列は、配列番号21に記載の配列を含む、請求項19に記載の方法。
- 前記第1の遺伝子改変マウスの生殖系列ゲノムおよび/または前記第2の遺伝子改変マウスの生殖系列ゲノムは、前記単一の再構成されたヒトVκ1−39/Jκ配列についてホモ接合性である、請求項15〜21のいずれか一項に記載の方法。
- 前記第1のヒト免疫グロブリン重鎖可変領域および/または前記第2のヒト免疫グロブリン重鎖可変領域が、ヒトV H 3−64またはその体細胞超変異バリアントを含む、請求項16〜22のいずれか一項に記載の方法。
- 前記抗原結合性タンパク質を作製することは、前記第1のマウスから選択された前記第1のヒト免疫グロブリン重鎖可変ドメインを含む第1の免疫グロブリン重鎖、および/または前記第2のマウスから選択された前記第2の免疫グロブリン重鎖可変ドメインを含む第2の免疫グロブリン重鎖を、再構成されたヒトVκ1−39/Jκ配列を含む免疫グロブリン軽鎖も発現する細胞において一緒に発現させることを含む、請求項15〜23のいずれか一項に記載の方法。
- 前記抗原結合性タンパク質は、完全ヒト抗原結合性タンパク質である、請求項1〜24のいずれか一項に記載の方法。
- 前記抗原結合性タンパク質は、二重特異的抗原結合性タンパク質である、請求項1〜25のいずれか一項に記載の方法。
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US30228210P | 2010-02-08 | 2010-02-08 | |
PCT/US2011/023971 WO2011097603A1 (en) | 2010-02-08 | 2011-02-08 | Common light chain mouse |
US13/412,936 US20120192300A1 (en) | 2010-02-08 | 2012-03-06 | Common Light Chain Mouse |
US13/412,936 | 2012-03-06 |
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CN101537180B (zh) | 2002-07-18 | 2016-02-10 | 莫鲁斯有限公司 | 抗体混合物的重组生产 |
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