JP6277213B2 - サルアデノウイルスベクターの増殖方法 - Google Patents
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Description
本出願は、2009年11月9日に提出された米国仮特許出願第61/259,343号の利益を主張する。この仮出願は、引用によりその全内容を本明細書の一部とする。
本明細書と同時に提出された、次に示す通りの、コンピュータで読み取り可能なヌクレオチド配列表は、引用によりその全内容を本明細書の一部とする:2010年11月8日に作成された「SEQ_Listing.TXT」というファイル名の38,463バイトのASCII(テキスト)ファイル1点。
本発明は、サルアデノウイルス、およびサルアデノウイルスを増殖させる方法であって、ヒトの疾患の治療および予防を含むさまざまな用途でのその使用を可能にする、該方法に関する。
この実施例は、サルアデノウイルスのプラーク形成が、ヒトアデノウイルスの要素E1およびE40RF6を発現するヒト細胞株上で非常に効率的であることを実証する。
この実施例は、ヒトアデノウイルス種のC因子を有するヒト細胞株での高力価のサルアデノウイルス子孫の発生を実証する。
この実施例は、ヒトアデノウイルスの要素を有するヒト細胞株のE1欠失を有するサルアデノウイルスベクターの構築と増殖を実証する。
SV−7:左手配列(配列番号:4);pPIX領域配列(配列番号:5);右配列(配列番号:6)。
SV−11:左手配列(配列番号:7);pPIX領域配列(配列番号:8);右配列(配列番号:9)。
SV−16:左手配列(配列番号:10);pPIX領域配列(配列番号:11);右配列(配列番号:12)。
SV−18:左手配列(配列番号:13);pPIX領域配列(配列番号:14);右配列(配列番号:15)。
SV−38:左手配列(配列番号:16);pPIX領域配列(配列番号:17);右配列(配列番号:18)。
SV−39:左手配列(配列番号:19);pPIX領域配列(配列番号:20);右配列(配列番号:21)。
この実施例は、E1領域配列の欠失があるサルアデノウイルスは複製能欠損型であることを実証する。
Claims (9)
- サルアデノウイルスをin vitroで増殖させる方法であって、
前記サルアデノウイルスをヒト細胞に接触させることを含み、
前記サルアデノウイルスは、ヒトアデノウイルスの1以上の遺伝子生成物をコードする核酸配列を含み、前記1以上の遺伝子生成物は、ヒト細胞における宿主の複製ブロックの軽減または抑制を担うE4領域の一部分であって、E4−ORF6を含み、かつ他のORFを含まないE4領域の一部分、によってコードされる遺伝子生成物を含み、前記サルアデノウイルスは、前記サルアデノウイルス由来の逆方向末端反復(ITR)配列を含み、前記ヒトアデノウイルス由来のITR配列を含まず、ここでサルとは新世界ザルおよび旧世界ザルの両方を指し、ヒト科のメンバーを含まず、それによって前記ヒト細胞において前記サルアデノウイルスが増殖される、方法。 - 前記サルアデノウイルスが、ヒトアデノウイルスC型遺伝子生成物をコードする核酸配列を含む、請求項1に記載の方法。
- 前記E4領域の一部分は、E4−ORF6である、請求項1または2に記載の方法。
- 前記細胞は、HEK−293細胞またはPerC.6細胞である、請求項1〜3のいずれか1項に記載の方法。
- 前記サルアデノウイルスは、複製能欠損型である、請求項1〜4のいずれか1項に記載の方法。
- 前記サルアデノウイルスは、増殖のためのサルアデノウイルスゲノムのE1A領域、E1B領域、およびE4領域のうちの1以上の相補性を必要とする、請求項1〜5のいずれか1項に記載の方法。
- 前記サルアデノウイルスは、サルアデノウイルスゲノムのE1領域の欠損およびE4領域の少なくとも一部の欠損を含む、請求項6に記載の方法。
- 前記1以上の遺伝子生成物は、ヒトアデノウイルスのE1A領域およびE1B領域の一方または両方にコードされた遺伝子生成物を含む、請求項1〜5のいずれか1項に記載の方法。
- 前記サルアデノウイルスは、サルアデノウイルスゲノムのE3領域の欠損をさらに含む、請求項1〜8のいずれか1項に記載の方法。
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Also Published As
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KR20120139672A (ko) | 2012-12-27 |
CN102770146B (zh) | 2016-02-03 |
EA201270607A1 (ru) | 2012-09-28 |
IL219492A0 (en) | 2012-06-28 |
JP2016146838A (ja) | 2016-08-18 |
EP2498791A2 (en) | 2012-09-19 |
CN102770146A (zh) | 2012-11-07 |
US9586998B2 (en) | 2017-03-07 |
US20120225470A1 (en) | 2012-09-06 |
MX2012005181A (es) | 2012-08-01 |
WO2011057248A2 (en) | 2011-05-12 |
EP2498791B1 (en) | 2014-12-24 |
CA2779632C (en) | 2019-08-20 |
AU2010314842A1 (en) | 2012-05-31 |
EP2853266A1 (en) | 2015-04-01 |
JP5980117B2 (ja) | 2016-08-31 |
US20150329834A1 (en) | 2015-11-19 |
JP2013509887A (ja) | 2013-03-21 |
WO2011057248A3 (en) | 2011-09-15 |
BR112012010824A2 (pt) | 2018-03-06 |
US9133248B2 (en) | 2015-09-15 |
EP2853266B1 (en) | 2018-01-31 |
CA2779632A1 (en) | 2011-05-12 |
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