Preparation of affinity latices and their application t of bioseparaton and cell activation.

亲和胶乳的制备及其在生物分离和细胞活化中的应用。

• 批准号: 05453214
• 负责人: KAWAGUCHI Haruma
• 金额: $ 3.97万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05453214/
• 关键词: Affinity latex; Bioseparator; Cell activator; Transcription factor; Membrane protein; Microsphere; ミクロスフェア; アフィニティ; ラテックス; スペーサー; DNA; RNA; RGDS; 転写制御; ホルモン

Either of biospecific couple was immobilizud to latex particles which were hydrophilic and have binding sites for the immobilization. Affinity latices thus prepared were employed for separation/purification of complementaey component, or cell activation via specific binding reaction. The following couples were used in this study ;a. (DNA) n E4TF1b.TFIID TFIIAc.Subunit of E4TF1 small nuclear Ribonucleo-Proteind.DNA Complementary DNAe.senseDNA antisenseRNAf.RGDS GPIIb/GPIIIaIn terms of f, RGDS-carrying affinity latex was used for two purposes ; GPIIb/IIIa purification from platelet membrane extract and leukocyte activation through RGD-GPIIb/IIIa interaction transferred to actin filament.The following three processed must be developed to get efficient affinity latices ; 1.Preparation of suitable carrier particle, 2.Immobilization of biocompounds in a proper mode, and 3.Search of suitable conditions to exhibit efficient biospecific reaction on particle surface.The effect of physical properties of particles on the efficiency of affinity latex was studied in detail.Polystirene-core/poly (qlycidyl methacrylate) shell particles were especially excellent among particles examined.In some systems, subunits or fragments of biofunctional molecules were immobilized on the particles.The resulting hybrid particles exhibited sufficient ability in bioseparation.Introduction of spacer chains between biocompound and particle surface improved the activity.

将两种生物特异性偶联物固定在具有亲水性和结合位点的乳胶颗粒上。由此制备的亲和胶乳用于分离/纯化互补组分，或通过特异性结合反应活化细胞。本研究中使用了以下夫妇;a. (DNA)n E4TF1b.TFIID TFIIAc.E4TF1小核核糖核蛋白亚基ind.DNA互补DNA e.正义DNA反义RNA f.RGDS GPIIb/GPIIIa携带RGDS的亲和胶乳有两个目的：从血小板膜提取物中纯化GPIIb/IIIa和通过RGD-GPIIb/IIIa相互作用将白细胞活化转移到肌动蛋白丝上。本文研究了聚苯乙烯核/聚苯乙烯核（甲基丙烯酸缩水甘油酯）壳颗粒在所检查的颗粒中是特别优异的。将生物功能分子的亚基或片段固定在颗粒上。所得的杂化颗粒表现出足够的能力，在生物化合物和颗粒表面之间引入间隔链提高了活性。

项目成果

Sumi, Y., et al.: "Application of cationic latex particles for protain separation." Colloids and Surfaces B : Biointerfaces.2. 419-427 (1994)

Sumi, Y. 等人：“阳离子乳胶颗粒在蛋白质分离中的应用。”

Urakami, Y., et al.: "Phagocytosis of microspheres with modified surfaces." Colloids and Surfaces B : Biointerfaces.3. 183-190 (1994)

Urakami, Y., et al.：“具有修饰表面的微球的吞噬作用。”

Inomata,Y.: "Purification of membrane receptors with pepitide-carrying affinity latex particles." Colloids and Surfaces B:Biointerface.(Accepted for publication).

Inomata，Y.：“用携带肽的亲和乳胶颗粒纯化膜受体。”

Y.Kasuya: "Activation of Human Neutrophil by Arg-Gly-Asp-Ser Immobilized on Microspheres" J.Biomed.Mater.Res.28. 397-404 (1994)

Y.Kasuya：“通过固定在微球上的 Arg-Gly-Asp-Ser 激活人类中性粒细胞”J.Biomed.Mater.Res.28。

H.Kawaguchi: "Bilomedical Applications of Polymeric Materials" CRC Press, 26 (1993)

H.Kawaguchi：“高分子材料的生物医学应用”CRC Press，26 (1993)