利用噬菌体随机肽库筛选抗柯萨奇病毒B3多肽的研究

本实验将柯萨奇病毒B3型(CVB3)大量扩增,应用蔗糖密度梯度离心法纯化病毒。利用噬菌体随机9肽库进行筛选,3轮淘洗后,测定噬菌体克隆抗病毒复制能力。提取阳性克隆DNA并进行测序,推导外源多肽的氨基酸序列。结果表明:3个具有明显抗病毒复制能力的噬菌体阳性克隆被筛选出来,使TCID50由10-7.5SFU/mL分别降至10-5.25、10-6、10-5.5SFU/mL,由此证明可以应用噬菌体肽库来筛选具有抗病毒作用的多肽,本研究为抗病毒多肽制剂的研究奠定了基础。

Anti-CVB3 Polypeptides Selected from A Phage Display Random Peptide Library

CVB3 was propagated in Hep-2 cells and purified by sucrose gradient centrifugation. The purified CVB3 reacted with random peptides library displaying 9 amino acids. After 3 rounds of screening, the capability of phage peptides in the inhibition of CVB3 replication was determined. DNA was extracted from the positive phage clones, sequenced and the amion acid sequence was deduced. The results showed that 3 phage positive peptides were identified and bound to CVB3 with high affinity. The binding of the peptide to CVB3 was shown to inhibit replication substantially. The TCID50 of progeny virus was reduced from 10-7.5SFU/mL to 10-5.25, 10-6, 10-5.5SFU/mL by the three peptides, respectively. The data demonstrate that peptides against CVB3 can be selected from phage peptide library and may prove useful as antiviral peptides agents.