Identification of a LNCaP-specific binding peptide using phage display.

Identification of a LNCaP-specific binding peptide using phage display.
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DOI:
10.1007/s11095-011-0469-7
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发表时间:
2011-10
影响因子:
3.7
通讯作者:
Cheng, Kun
Cheng, Kun
中科院分区:
医学3区
文献类型:
--
作者:
Qin, Bin;Tai, Wanyi;Shukla, Ravi S.;Cheng, Kun

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使用噬菌体展示库鉴定 LNCaP 特异性肽并评估其在靶向药物递送中的潜在应用。通过细胞噬菌体 ELISA 评估所选噬菌体的结合能力。合成由最特异的噬菌体克隆编码的 KYL 肽,用荧光素标记,并在各种细胞系中进行测定。合成了由KYL肽和促凋亡肽d(KLAKLAK)2组成的融合肽,并在不同细胞上评估细胞死亡效果。此外,KYL 肽与阳离子蛋白鱼精蛋白缀合,以探索其在 siRNA 递送中的潜在应用。鉴定出一种与 LNCaP 细胞具有高结合亲和力的噬菌体克隆。细胞噬菌体 ELISA 和免疫染色证明该噬菌体对 LNCaP 细胞具有高度特异性。与其他细胞相比,荧光素标记的 KYL 肽与 LNCaP 细胞具有更高的结合力。由KYL肽和促凋亡肽组成的融合肽在LNCaP细胞中诱导细胞死亡,但在PC-3细胞中不诱导细胞死亡。 KYL 肽-鱼精蛋白缀合物还可有效地将荧光素标记的 siRNA 递送至 LNCaP 细胞中。我们鉴定了一种 LNCaP 特异性肽,并证明了其在向 LNCaP 细胞靶向给药方面的潜在应用。
To identify a LNCaP-specific peptide using a phage display library and evaluate its potential applications in targeted drug delivery. Binding abilities of selected phages were evaluated by cell phage ELISA. The KYL peptide encoded by the most specific phage clone was synthesized, labeled with fluorescein, and assayed in various cell lines. A fusion peptide composed of the KYL peptide and a proapoptotic peptide d(KLAKLAK)2 was synthesized, and the cell death effect was evaluated on different cells. Moreover, the KYL peptide was conjugated to a cationic protein, protamine, to explore its potential application in siRNA delivery. One phage clone with a high binding affinity to LNCaP cells was identified. Cell phage ELISA and immunostaining demonstrated high specificity of this phage to LNCaP cells. The fluorescein-labeled KYL peptide exhibited higher binding to LNCaP cells in comparison to other cells. The fusion peptide composed of the KYL peptide and the proapoptotic peptide induced cell death in LNCaP cells, but not in PC-3 cells. The KYL peptide-protamine conjugate also efficiently delivered a fluorescein-labeled siRNA into LNCaP cells. We identified a LNCaP-specific peptide and demonstrated its potential applications in targeted drug delivery to LNCaP cells.
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