The use of N-terminal immobilization of PTH(1-34) on PLGA to enhance bioactivity.
The use of N-terminal immobilization of PTH(1-34) on PLGA to enhance bioactivity.
复制标题
利用 PTH(1-34) 的 N 端固定在 PLGA 上来增强生物活性。
DOI：
10.1016/j.biomaterials.2008.04.019
复制
发表时间：
2008
期刊：
影响因子：
14
通讯作者：
Puleo,DavidA
中科院分区：
文献类型：
--
作者：
Sharon,JessicaL;Puleo,DavidA
这项工作的目的是控制生物活性分子的方向固定在一个可生物降解的基板上，以提高其可访问性结合细胞表面受体，因此，增加生物活性。骨肽，甲状旁腺激素（1-34）（PTH（1-34）），被用来证明的方法。为此，内在的N-末端丝氨酸残基被氧化以产生醛基，该醛基在中性条件下特异性结合至酰肼衍生的聚（丙交酯-共-乙交酯）以形成腙键。与简单吸附或直接随机附着相比，使用二酰肼间隔物显著增加了固定化肽的量。在探测固定的PTH（1-34）的可及性时，使用较长的二酰肼间隔物的连接增强了抗体对肽的N-末端区域中的表位的结合。最长的间隔区也增加了C-末端抗体的结合。此外，通过间隔物连接肽的底物刺激细胞内cAMP的合成，活性随二酰肼长度增加。使用最长间隔基固定的PTH（1-34）比随机结合和吸附显著更有效。生物活性肽与表面的定点结合呈现用于与细胞结合的生物分子，从而增强与受体的相互作用，因此该方法可用于获得优选的局部组织反应。
The objective of this work was to control the orientation of bioactive molecules immobilized on a biodegradable substrate to improve their accessibility for binding to cell surface receptors and, therefore, to increase bioactivity. The osteotropic peptide, parathyroid hormone (1–34) (PTH(1–34)), was used to demonstrate the approach. To this end, the intrinsic N-terminal serine residue was oxidized to create an aldehyde group that specifically bound to hydrazide-derivatized poly(lactide-co-glycolide) under neutral conditions to form a hydrazone bond. Use of dihydrazide spacers significantly increased the amount of peptide immobilized compared to simple adsorption or direct, random attachment. In probing accessibility of immobilized PTH(1–34), attachment using longer dihydrazide spacers enhanced binding of an antibody against an epitope in the N-terminal region of the peptide. The longest spacer also increased binding of a C-terminal antibody. Furthermore, substrates with peptide tethered via spacers stimulated intracellular synthesis of cAMP, with activity increasing with dihydrazide length. PTH(1–34) immobilized using the longest spacer was significantly more effective than both random binding and adsorption. Site-directed binding of bioactive peptides to surfaces presents biomolecules for binding with cells so as to enhance interaction with receptors, and therefore the approach may be useful for obtaining preferred localized tissue responses.