JP6778380B2 - 吸着材 - Google Patents
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- JP6778380B2 JP6778380B2 JP2016162834A JP2016162834A JP6778380B2 JP 6778380 B2 JP6778380 B2 JP 6778380B2 JP 2016162834 A JP2016162834 A JP 2016162834A JP 2016162834 A JP2016162834 A JP 2016162834A JP 6778380 B2 JP6778380 B2 JP 6778380B2
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Description
水中でイオン化し得る官能基を複数有する高分子材料としてポリアミンを用いた。具体的には、標的物質をヒトIgG抗体とし、該高分子材料としてε−ポリリジン(EPL)を用い、吸着部位を(4−ヒドロキシフェニルエチルアミノ)(フェニルアミノ)トリアジン(ApA)とした。なお、ApAは、抗体吸着能を有するプロテインA模倣分子である。
標的物質をヒトIgG抗体とし、高分子材料としてEPLを用い、吸着部位を、(3−チア−5−ピリジルペンチル)ビニルスルホン(MEP−DVS)由来の(3−チア−5−ピリジルペンチル)スルホニルエチル部分とした。
種々の高分子材料と吸着部位の組み合わせを用いて実施例3−6の吸着材を得た。具体的には、表1に示す高分子材料と、吸着部位を形成するための化合物(1〜20当量)とを混合し、所定の条件にて反応させてポリマー3−6を合成し、得られた各ポリマーを実施例1と同様にして担体に固定化して、実施例3−6の吸着材を得た。
高分子材料としてEPLを用い、吸着部位をApAとした。実施例1におけるポリマー1の担体への固定化と同様にして、ApAを、NHSエステル基を表面に有する架橋アガロース担体に固定化し、その後、EPLを担体に固定化して、APAおよびEPLが別々に担体に固定化された実施例7の吸着材を得た。
架橋アガロース担体に、NaOH存在下でジビニルスルホン(DVS)を作用させた後に2−ピリジルエタンチオール(MEP)塩酸塩を作用させることで、MEP−DVSに由来する(3−チア−5−ピリジルペンチル)スルホニルエチル部分が担体に導入された比較例1の吸着材を得た。
表面にアミノ基を有する架橋アガロース担体に、プロテインA模倣分子22/8の原料であるクロロトリアジン誘導体のDMF溶液を作用させることで、プロテインA模倣分子22/8が担体に導入された比較例2の吸着材を得た。
架橋アガロース担体にヒトIgG抗体を固定化して、ヒトIgG抗体が担体に導入された比較例3の吸着材を得た。
NHSエステル基を表面に有する架橋アガロース担体にポリアミドアミンデンドリマーの0.1重量%溶液を加え、得られた懸濁液を室温で一晩振盪することで、担体にポリアミドアミンデンドリマーが固定化された比較例4の吸着材を得た。
架橋アガロース担体を比較例5の吸着材とした。
実施例1−7および比較例1−5の吸着材について、温度変化に伴うイオン化率の変化を測定した。
実施例1−7の吸着材および比較例1−5の吸着材の温度変化に伴う標的物質に対する吸着特性の変化をカラム中で測定した。
実施例1−7の吸着材および比較例1−5の吸着材の温度変化に伴う標的物質に対する吸着特性の変化を、吸着材を懸濁液の状態で用いるビーズ分散系で測定した。
20 吸着材
1 高分子材料
2 水中でイオン化し得る官能基
3 高分子主鎖
4 吸着部位
5 標的物質
6 担体
Claims (10)
- 水中でイオン化し得る官能基を複数有する、下限臨界溶解温度を示さない高分子材料と、
タンパクと分子間相互作用により吸着可能な吸着部位と、
担体と
を備えることを特徴とするタンパク用吸着材であって、
前記高分子材料が、ポリリジン、アミノ基を複数有するデンドリマー、イミノ基を複数有するデンドリマー、ポリエチレンイミン、ポリアリルアミン、ポリグルタミン酸、またはそれらを部分構造として含む高分子材料であり、
前記吸着部位が、プロテインA部分構造または模倣分子、プロテインG部分構造または模倣分子、プロテインL部分構造または模倣分子、糖鎖結合分子、チオフィリックリガンドおよび抗原または抗原部分構造から選ばれる少なくとも1つであり、
前記高分子材料と前記吸着部位が直接結合しているか、または前記高分子材料と前記吸着部位が結合しておらず、異なる位置で前記担体に固定化されており、
前記高分子材料と前記吸着部位が直接結合している場合であって、前記高分子材料がポリリジンである場合、前記吸着部位がポリリジンの側鎖に結合している、
タンパク用吸着材。 - 外部環境の変化によって高分子材料のイオン化率が変化することを特徴とする、請求項1に記載のタンパク用吸着材。
- 外部環境の変化が、吸着材が接触している溶液の温度、塩濃度、誘電率またはpHの変化であることを特徴とする、請求項2に記載のタンパク用吸着材。
- 前記ポリリジンがε−ポリリジンであることを特徴とする、請求項1に記載のタンパク用吸着材。
- タンパクが抗体であることを特徴とする、請求項1に記載のタンパク用吸着材。
- 担体またはその表面が、多糖類、架橋された多糖類、ポリスチレン、シリカまたは金属酸化物を有することを特徴とする、請求項1に記載のタンパク用吸着材。
- 担体またはその表面が、多糖類または架橋された多糖類を有することを特徴とする、請求項6に記載のタンパク用吸着材。
- 請求項1に記載のタンパク用吸着材にタンパクを含む溶液を接触させる工程と、
その後請求項3に記載の外部環境の変化を印加する工程と
を含むタンパクの精製方法。 - 請求項1に記載のタンパク用吸着材が充填されたカラムを用いる、請求項8に記載の精製方法。
- 請求項1に記載のタンパク用吸着材を懸濁液の状態で用いる、請求項8に記載の精製方法。
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
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JP2016162834A JP6778380B2 (ja) | 2016-08-23 | 2016-08-23 | 吸着材 |
CN201780029791.2A CN109153006A (zh) | 2016-08-23 | 2017-07-10 | 吸附材料 |
US16/301,303 US10898878B2 (en) | 2016-08-23 | 2017-07-10 | Adsorbent material |
EP17843229.0A EP3505240A4 (en) | 2016-08-23 | 2017-07-10 | ADSORBENT MATERIAL |
PCT/JP2017/025139 WO2018037742A1 (ja) | 2016-08-23 | 2017-07-10 | 吸着材 |
TW106125747A TWI660774B (zh) | 2016-08-23 | 2017-07-31 | 吸附材料 |
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CN109225149B (zh) * | 2018-10-09 | 2021-02-05 | 齐齐哈尔大学 | 一种TiO2改性壳聚糖吸附剂及其制备方法和应用 |
CN111408354B (zh) * | 2020-03-30 | 2022-11-25 | 杭州师范大学钱江学院 | 一种基于生物纤维面膜边角料的固相萃取剂及其制备方法 |
JPWO2022039112A1 (ja) * | 2020-08-20 | 2022-02-24 | ||
CN114505077B (zh) * | 2022-01-28 | 2023-09-08 | 湖南城市学院 | 一种处理污水用复合催化剂及制备方法 |
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JP3254815B2 (ja) | 1993-05-13 | 2002-02-12 | 富士レビオ株式会社 | 凝集反応測定用粒子の製造方法 |
JP3441496B2 (ja) | 1993-11-17 | 2003-09-02 | 株式会社セルシード | アフィニティー分離材料 |
EP0851768B1 (en) | 1995-09-01 | 2002-04-24 | University of Washington | Interactive molecular conjugates |
AU4612400A (en) | 1999-05-11 | 2000-11-21 | Japan Chemical Innovation Institute | Affinity-controlling material with the use of stimulus-responsive polymer and separation/purification method with the use of the material |
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US8969353B2 (en) * | 2008-11-07 | 2015-03-03 | Massachusetts Institute Of Technology | Aminoalcohol lipidoids and uses thereof |
JP4922341B2 (ja) * | 2009-05-14 | 2012-04-25 | シャープ株式会社 | 標的認識分子および標的認識分子を固定化する方法 |
CN102580685B (zh) | 2011-12-15 | 2013-05-29 | 清华大学 | 一种多孔的多糖类细胞吸附材料及其制备方法与应用 |
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JP6169885B2 (ja) | 2013-05-07 | 2017-07-26 | 株式会社日立製作所 | 精製装置及び精製方法 |
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TW201806669A (zh) | 2018-03-01 |
WO2018037742A1 (ja) | 2018-03-01 |
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EP3505240A1 (en) | 2019-07-03 |
TWI660774B (zh) | 2019-06-01 |
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