Yao Shasha, Huang Yanqin, Zhao Yanbao, Zhang Yu, Zou Xueyan, Song Chunpeng
Key Laboratory for Special Functional Materials, Henan University, Kaifeng 475004, PR China.
Key Laboratory of Plant Stress Biology, Henan University, Kaifeng 475004, PR China.
Mater Sci Eng C Mater Biol Appl. 2014 Jun 1;39:1-5. doi: 10.1016/j.msec.2014.02.034. Epub 2014 Feb 22.
A simple strategy has been developed to synthesize hydroxyapatite (HAP) nanoparticles (NPs) in a simulated body fluid (SBF). The HAP NPs have an average diameter of 50nm and present porous structure. By taking advantage of surface hydroxyl groups, the HAP NPs are further modified with iminodiacetic acid (IDA), followed by chelating Ni(2+) ions. The HAP/IDA-Ni(2+) NPs as novel adsorbent can capture directly histidine-tagged (His-tagged) proteins from the mixture of lysed cells without sample pretreatment. Results indicated that the HAP/IDA-Ni(2+) NPs present negligible nonspecific adsorption and high protein binding ability, and their specificity and affinity toward His-tagged proteins can remain after 5 times of recycling. The HAP/IDA-Ni(2+) NPs are especially suitable for purification of His-tagged proteins with low molecule weight.
已经开发出一种简单的策略，用于在模拟体液（SBF）中合成羟基磷灰石（HAP）纳米颗粒（NPs）。HAP NPs的平均直径为50nm，并呈现多孔结构。利用表面羟基，HAP NPs用亚氨基二乙酸（IDA）进一步修饰，然后螯合Ni(2+)离子。作为新型吸附剂的HAP/IDA-Ni(2+) NPs无需样品预处理即可直接从裂解细胞混合物中捕获组氨酸标签（His-tagged）蛋白。结果表明，HAP/IDA-Ni(2+) NPs的非特异性吸附可忽略不计，且具有高蛋白结合能力，并且它们对His-tagged蛋白的特异性和亲和力在5次循环后仍可保持。HAP/IDA-Ni(2+) NPs特别适用于纯化低分子量的His-tagged蛋白。
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