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基于聚(N-甲基丙烯酰基-L-亮氨酸)的温敏肽模拟涂层:性能、蛋白质吸附和细胞生长。

Temperature-responsive peptide-mimetic coating based on poly(N-methacryloyl-l-leucine): properties, protein adsorption and cell growth.

机构信息

Smoluchowski Institute of Physics, Jagiellonian University, Reymonta 4, 30-059 Kraków, Poland.

"Lvivska Polytechnika" National University, S. Bandery 12, 79013 Lviv, Ukraine.

出版信息

Colloids Surf B Biointerfaces. 2014 Jun 1;118:270-9. doi: 10.1016/j.colsurfb.2014.03.049. Epub 2014 Apr 6.

Abstract

Poly(N-methacryloyl-l-leucine) (PNML) coatings were successfully fabricated via polymerization from peroxide initiator grafted to premodified glass substrate. Chemical composition and thickness of PNML coatings were determined using time of flight-secondary ion mass spectrometry (TOF- SIMS) and ellipsometry, respectively. PNML coatings exhibit thermal response of the wettability, between 4 and 28°C, which indicates a transition between hydrated loose coils and hydrophobic collapsed chains. Morphology of the PNML coating was observed with the AFM, transforming with increasing temperature from initially relatively smooth surface to rough and more structured surface. Protein adsorption observed by fluorescence microscopy for model proteins (bovine serum albumin and lentil lectin labeled with fluorescein isothiocyanate) at transition from 5 to 25°C, showed high affinity of PNML coating to proteins at all investigated temperatures and pH. Thus, PNML coating have significant potential for medical and biotechnological applications as protein capture agents or functional replacements of antibodies ("plastic antibodies"). The high proliferation growth of the human embryonic kidney cell (HEK 293) onto PNML coating was demonstrated, indicating its excellent cytocompatibility.

摘要

聚(N-甲基丙烯酰基-L-亮氨酸)(PNML)涂层通过过氧化物引发剂接枝到预改性玻璃基底上的聚合成功制备。使用飞行时间-二次离子质谱(TOF-SIMS)和椭圆光度法分别确定了 PNML 涂层的化学组成和厚度。PNML 涂层在 4 到 28°C 之间表现出润湿性的热响应,表明在水合松散线圈和疏水性塌陷链之间存在转变。通过原子力显微镜(AFM)观察 PNML 涂层的形貌,随着温度的升高,从最初相对光滑的表面转变为粗糙且更具结构的表面。通过荧光显微镜观察模型蛋白(用异硫氰酸荧光素标记的牛血清白蛋白和扁豆凝集素)在从 5°C 到 25°C 的转变过程中的吸附,表明 PNML 涂层在所有研究温度和 pH 值下对蛋白质均具有高亲和力。因此,PNML 涂层作为蛋白质捕获剂或抗体的功能替代品(“塑料抗体”)在医学和生物技术应用中具有重要的应用潜力。PNML 涂层上的人胚肾细胞(HEK 293)的高增殖生长得到了证明,表明其具有优异的细胞相容性。

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