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带有接枝聚合物“触手”的超滤膜表面:共价蛋白质结合的制备、表征及应用

Ultrafiltration membrane surfaces with grafted polymer 'tentacles': preparation, characterization and application for covalent protein binding.

作者信息

Ulbricht M, Riedel M

机构信息

Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, Institut für Chemie, BR Deutschland.

出版信息

Biomaterials. 1998 Jul;19(14):1229-37. doi: 10.1016/s0142-9612(98)00029-5.

DOI:10.1016/s0142-9612(98)00029-5
PMID:9720886
Abstract

Ultrafiltration (UF) membranes from polysulfone (PSf) were functionalized by heterogeneous photo-initiated graft copolymerization of acrylic acid (AA). With radiation susceptible PSf, only proper selection of the UV energy (lambda > 350 nm; for selective excitation of the photoinitiator) yielded membranes with preserved UF barrier layer. Possibilities for adjusting structure and morphology of the graft polymer (g-PAA) layer by variation of functionalization parameters such as AA concentration and UV irradiation time were investigated. Very long grafted chains (Mw > 10(5) g mol(-1)) at varied grafting density (GD = 0.01 ... 1.2 nmol cm(-2), relative to the outer surface area) were obtained. Partial penetration of the UF barrier layer by g-PAA was verified. Covalent immobilization of bovine serum albumin (BSA), gamma-globulin (gamma-Gl) and alkaline phosphatase (APh) was achieved by coupling with a water soluble carbodiimide. Bound BSA and gamma-Gl amounts were up to gamma = 10 microg cm(-2), for membranes accessible only from the outer surface thus not using the entire pore volume. Locally addressed covalent protein immobilization after photo-patterning the PSf surface could be visualized with a fluorescent FITC-BSA conjugate. A strong salt effect onto immobilized APh activity (increase with NaCl concentration) was observed, indicating internal transport/accessibility limitations in the g-PAA layer. Correlations between PAA structure (Mw, GD) and accessibility (from BSA or gamma-G1 binding and APh activity) could be established. The 'tentacle' g-PAA functionalized PSf UF membranes having preserved UF barrier and, e.g., with surface-bound receptors will find application in cell cultures under diffusion or perfusion conditions.

摘要

通过丙烯酸(AA)的非均相光引发接枝共聚反应对聚砜(PSf)超滤(UF)膜进行功能化改性。对于对辐射敏感的PSf,只有适当选择紫外光能量(λ>350 nm;用于光引发剂的选择性激发)才能得到具有完整超滤阻挡层的膜。研究了通过改变功能化参数(如AA浓度和紫外照射时间)来调整接枝聚合物(g-PAA)层结构和形态的可能性。在不同的接枝密度(GD = 0.01...1.2 nmol cm-2,相对于外表面面积)下获得了非常长的接枝链(Mw>105 g mol-1)。证实了g-PAA对超滤阻挡层的部分渗透。通过与水溶性碳二亚胺偶联实现了牛血清白蛋白(BSA)、γ-球蛋白(γ-Gl)和碱性磷酸酶(APh)的共价固定。对于仅从外表面可接触的膜,即不使用整个孔体积的膜,结合的BSA和γ-Gl量高达γ = 10 μg cm-2。用荧光FITC-BSA缀合物可以观察到在PSf表面进行光图案化后局部定位的共价蛋白质固定。观察到盐对固定化APh活性有强烈影响(随NaCl浓度增加),表明g-PAA层存在内部传输/可及性限制。可以建立PAA结构(Mw,GD)与可及性(来自BSA或γ-G1结合以及APh活性)之间的相关性。具有完整超滤阻挡层且例如带有表面结合受体的“触手”状g-PAA功能化PSf超滤膜将在扩散或灌注条件下的细胞培养中得到应用。

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