含Arg-Gly-Asp肽与仿生PLGA-(ASP-PEG)材料结合的实验研究

设计合成含Arg-Gly-AspArg(R)-精氨酸,Gly(G)-甘氨酸,Asp(D)-天冬氨酸]的非病毒基因载体的肽(K16-GRGDSPC) (K-赖氨酸,S-丝氨酸,P-脯氨酸,C-半胱氨酸),用其作材料聚(丙交酯-co-乙交酯)-(天冬氨酸-聚乙二醇)交替预聚物PLGA-(ASP-PEG)]的表面修饰.合成K16-GRGDSPC, 将PLGA-(ASP-PEG)制成A、B、C三块薄片.薄片 A 、B与交联剂反应,反应后的薄片A再与肽反应,薄片 C作为对照.质谱仪(MS)、高效液相色谱分析仪(HPLC)检测肽的分子量及纯度;x线光电子能谱法(XPS)检测材料表面硫元素;分光光度仪检测残液未反应肽含量.HPLC检测肽的纯度为94.13%,MS检测肽分子量为2741.26.XPS检测改性薄片A中硫元素结合能为164 eV,碳、硫元素的含量比值(C/S)为99.746:0.1014;反应后薄片 B中硫元素结合能为162 eV与 164 eV,C/S为99.574:0.4255; 薄片 C中无硫元素.残液管OD值为0.069, 薄片A表面接枝肽密度为0.04 mg/mm2.提示通过交联剂可将所合成的肽K16-GRGDSPC结合到仿生材料PLGA-(ASP-PEG)表面.

Arg-Gly-Asp-containing Peptide Combining with the Biomimetic And Modified PLGA-(ASP-PEG)

Arg-Gly-Asp-(RGD) containing peptide characterized as the non-viral gene vector was synthesized to modify the surface of PLGA-(ASP-PEG). The Peptide (K16-GRGDSPC) was synthesized. PLGA-(ASP-PEG) was executed into chips A, B and C. Chip C was regarded as control. Chips A and B reacted with the cross-linker, then Chip A reacted with peptide. Mass spectrometry (MS) and high performance liquid chromatography (HPLC) detected the molecular weight and the purity of peptide. Sulphur in the surface of materials was detected by X-ray photoelectron spectrometry (XPS). The peptide content in the residual solution was detected by Spectrometer. HPLC showed the peptide purity was 94.13%; MS showed the molecular weight was 2741.26. XPS revealed the binding energy of the sulphur in reacted Chip A was 164 eV in reacted Chip B, 164eV and 162 eV; the ratios of carbon to sulphur in reacted Chip A and B were 99.746:0.1014 and 99.574:0.4255, respectively. There was no sulphur in Chip C. The optical density value (OD) of the resident solution was 0.069. The peptide density of reacted Chip A was 0.04 mg/mm2. The peptide was manufactured and linked to the surface of the biomimetic PLGA-(ASP-PEG) with the cross-linker.