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目的获得与hERG钾通道特异性结合的功能性亲和肽。方法以hERG1a亚基第3个细胞外环(L3)为靶蛋白分子,对噬菌体随机七肽库进行亲和筛选,ELISA法检测噬菌体克隆与靶蛋白的亲和性,使用全自动膜片钳技术观察亲和肽的活性。结果与结论经过3轮亲和筛选后,随机挑选15个噬菌体克隆对其亲和性做ELISA鉴定,均显示较强的阳性结果。将上述阳性克隆进行DNA测序得到3种不同小肽序列。电生理学研究表明,合成的3个小肽均能显著抑制hERG钾电流,为靶向hERG钾通道抗肿瘤研究提供了新的研究思路。
Objective To obtain a functional affinity peptide that specifically binds to the hERG potassium channel. METHODS: The third extracellular loop (L3) of hERG1a subunit was used as the target protein molecule to screen the phage random heptapeptide library. The affinity of the phage clone to the target protein was detected by ELISA and the automatic patch clamp technique was used. Observe the activity of the affinity peptide. Results and Conclusions After 3 rounds of affinity screening, 15 phage clones were randomly selected for their affinity for ELISA identification, all showed strong positive results. The above positive clones were subjected to DNA sequencing to obtain 3 different small peptide sequences. Electrophysiological studies have shown that the synthesis of three small peptides can significantly inhibit the hERG potassium current, providing a new research idea for targeting hERG potassium channel anti-tumor research.