含IKVAV肽自组装成凝胶的实验

IKVAV-containing peptide self-assembly into hydrogel

目的:利用特定结构的多肽在生理条件下的自组装特性构筑功能性再生支架,对神经修复和再生具有重要意义,并可望突破其他支架材料的局限性。设计合成含IKVAV肽(C16H31O-AAAAGGGEIKVAV),并诱发其自组装过程。方法:实验于2005-01/2006-07在华中科技大学同济医学院附属协和医院骨科实验室完成。实验方法:①寡肽的合成:采用固相法合成肽。②寡肽自组装:取质量分数为0.01的寡肽溶液150μL,置于玻璃小瓶中,pH值调整为9.5,然后滴加等体积的磷酸盐缓冲液,震荡离心(2000r/min,离心10min)。实验评估:①高效液相色谱仪测肽纯度;色谱仪测肽的相对分子质量。②透射电镜下观察自组装凝胶超微结构。结果:①高效液相色谱仪示多肽纯度为95%,质谱仪示肽的相对分子质量为1351.6。②质量分数为0.01的肽溶液在磷酸盐缓冲液触发下数秒钟形成凝胶,透射电镜示自组装的凝胶为交织成网状的纤维结构,直径3～6nm,长度100～1500nm。结论:实验合成了含IKVAV肽,在磷酸盐缓冲液中可自组装成多孔凝胶结构。

AIM： To construct functional regeneration scaffold characterized by self-assembly of peptide with physiological specific structure is significant for repair and regeneration of nerve. It is hoped to break through the limitation of other scaffolds. This article is aimed to synthesize IKVAV-containing （C16H31-AAAA GGGEIKVAV） peptide and induce self-assembly. METHODS： The experiment was performed at the Laboratory of Department of Orthopaedics, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology from January 2005 to July 2006. Experimental methods： ①The peptide was synthesized in solid-phase way. ②Oligopeptide self-assembly： 150 μL oligopeptide solution of 0.01 mass fraction was put in glass flask where pH was equal to 9.5 with the addition of phosphate buffered solution （PBS） of the same volume, and the solution was shaken and ceotrifugalized at 2 000 r/min for 10 minutes. Experimental evaluation： ①Peptide concentration was detected with high performance liquid chromatogram （HPLC）. Relative molecular mass （RMM） of peptide was determined with chromatography. ② Ultramicrostructure of hydrogel was determined with transmission electron microscope （TEM）. RESULTS： ①Purity and RMM were 95% and 1 351.6, respectively identified with HPLC. ② Peptide of 0.01 mass fraction was self-supported into hydrogel in few seconds in PBS solution. TEM showed that self-assembly hydrogel was consisted of the interconnected nanofibers, which varied from 3 nm to 6 nm in diameter and 100 nm to 1 500 nm in length. CONCLUSION： The IKVAV-containing peptide is synthesized and self organized successfully into porous hydrogel, which is triggered with PBS solution.