Weeks C Andrew, Aden Bethany, Kilbey S Michael, Janorkar Amol V
Department of Biomedical Materials Science, School of Dentistry, University of Mississippi Medical Center, 2500 North State Street, Jackson, Mississippi 39216, United States.
Departments of Chemistry and Chemical and Biomolecular Engineering, University of Tennessee, 322 Buehler Hall, 1420 Circle Drive, Knoxville, Tennessee 37996, United States.
ACS Biomater Sci Eng. 2016 Dec 12;2(12):2196-2206. doi: 10.1021/acsbiomaterials.6b00398. Epub 2016 Nov 16.
Many structural variants of elastin-like polypeptides (ELPs), the genetically engineered equivalents of part of human elastin, currently are being investigated for drug delivery and tissue engineering. Here, we report preparation of six different aminated ELP conjugates via two strategies. In the first, a direct linking strategy was used to couple hydrophobic ELP with either polyethyleneimine, polylysine, or polyarginine. In the second, conjugates were made by attaching ELP onto the reactive polymer, poly(2-vinyl-4,4-dimethyl azlactone), and then exhaustively reacting residual azlactone groups with either ethylenediamine, 1,4-butanediamine, or arginine. Molecular size and chemistry of the resulting six aminated-ELP conjugates were confirmed through gel electrophoresis, FTIR spectroscopy, and mass spectrometry. Dynamic light scattering analysis showed that the conjugates prepared using the "direct reaction scheme" formed small aggregates as well as retained their inverse volume-phase transition temperature behavior. The conjugates prepared using the "reactive polymer linker scheme" also retained this transition temperature behavior. -Phthalaldehyde assay was used to measure the relative primary amine content of the ELP conjugates. Overall, we prepared an array of aminated-ELPs with independently varying amine content and chemistry (i.e., the same amine content for different materials and different amine contents for the same material). Synthesis of such amphiphilic ELP structures that otherwise cannot be prepared through genetic engineering has the potential to further extend the versatility of the ELPs for many biomedical applications.
弹性蛋白样多肽(ELP)是人类弹性蛋白部分的基因工程等同物,目前其许多结构变体正在用于药物递送和组织工程的研究。在此,我们报告了通过两种策略制备六种不同的胺化ELP缀合物。第一种方法是采用直接连接策略,将疏水性ELP与聚乙烯亚胺、聚赖氨酸或聚精氨酸偶联。第二种方法是将ELP连接到反应性聚合物聚(2-乙烯基-4,4-二甲基氮杂环丁烷酮)上,然后使残留的氮杂环丁烷酮基团与乙二胺、1,4-丁二胺或精氨酸充分反应,从而制备缀合物。通过凝胶电泳、傅里叶变换红外光谱(FTIR)和质谱对所得六种胺化ELP缀合物的分子大小和化学组成进行了确认。动态光散射分析表明,采用“直接反应方案”制备的缀合物形成了小聚集体,并且保留了其反相体积转变温度行为。采用“反应性聚合物连接方案”制备的缀合物也保留了这种转变温度行为。采用邻苯二甲醛分析法测量ELP缀合物的相对伯胺含量。总体而言,我们制备了一系列胺化ELP,其胺含量和化学组成独立变化(即不同材料的胺含量相同,同一材料的胺含量不同)。合成这种无法通过基因工程制备的两亲性ELP结构有可能进一步扩展ELP在许多生物医学应用中的多功能性。