Isolation of a Mycoplasma-specific binding peptide from an unbiased phage-displayed peptide library
Isolation of a Mycoplasma-specific binding peptide from an unbiased phage-displayed peptide library
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DOI：
10.1039/b504572j
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发表时间：
2005-07-01
影响因子：
--
通讯作者：
Brown, KC
中科院分区：
文献类型：
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作者：
De, J;Chang, YC;Brown, KC
医学的一个重要目标是开发将治疗分子特异性靶向病原体或病原体感染细胞的方法。然而，在针对细菌感染细胞的细胞特异性靶向方面进展甚微。使用噬菌体展示方法，我们分离出了一种 20 聚体肽，该肽可与组织培养中精氨酸支原体感染的胰腺 β 细胞结合。该肽与精氨酸微球菌感染的 β 细胞的结合比对照噬菌体强 200 倍，并且对受感染的细胞具有特异性。此外，将精氨酸支原体污染转移到另一个细胞系会使新感染的细胞系对肽结合敏感。免疫定位实验表明，该肽与粘附在细胞表面的精氨酸微球菌结合。游离的合成肽在没有噬菌体载体的情况下保留其结合，并且肽的四聚化增加了其对受感染细胞的亲和力。人们已经努力使用这种肽通过涂有所选肽的铁磁珠来消除受感染细胞系中的支原体。通过这种方法，一次分割即可将感染减少十倍。我们的结果表明，从无偏选择中分离出来的这种肽可能可用于检测和减少培养细胞中的支原体感染。此外，我们的研究结果的一般含义是，噬菌体展示方法可能有助于识别以特定方式靶向多种其他生物病原体的肽。
An important goal in medicine is the development of methods for cell-specific targeting of therapeutic molecules to pathogens or pathogen-infected cells. However, little progress has been made in cell-specific targeting of bacterially infected cells. Using a phage display approach, we have isolated a 20-mer peptide that binds to Mycoplasma arginini infected pancreatic beta-cells in tissue culture. This peptide binds to M. arginini infected beta-cells 200 times better than a control phage and is specific for the infected cells. Furthermore, transferring the M. arginini contamination to another cell line renders the newly infected cell line susceptible to peptide binding. Immunolocalization experiments suggest that the peptide is binding to M. arginini adhered to the cell surface. The free synthetic peptide retains its binding in the absence of the phage vehicle and tetramerization of the peptide increases its affinity for the infected cells. Efforts have been made to use this peptide to eliminate Mycoplasma from infected cell lines using ferromagnetic beads coated with the selected peptide. A ten-fold reduction of infection was accomplished with one fractionation via this approach. Our results suggest that this peptide, isolated from an unbiased selection, may be of utility for the detection and reduction of Mycoplasma infection in cultured cells. Furthermore, a general implication of our findings is that phage display methods may be useful for identifying peptides that target a broad array of other biological pathogens in a specific fashion.