Dan Zhang1, Huan Jia2, Yan Wang3, Wei-Ming Li4, Ying-Chun Hou5, Shi-Wei Yin6, Thomas D Wang7, Shui-Xiang He8, Shao-Ying Lu9. 1. Department of Gastroenterology, The First Affiliated Hospital of Medical School, Xian Jiaotong University, Xi'an, 710061, China. zhangdan.124@stu.xjtu.edu.cn. 2. Department of General Surgery, The First Affiliated Hospital of Xi'an Medical University, Xi'an, 710077, Shaanxi, China. aerogle@stu.xjtu.edu.cn. 3. Institute for Biomedical Sciences of Pain, Tangdu Hospital, The Fourth Military Medical University, Xi'an, 710038, Shaanxi, China. wangxiaoyan2148@163.com. 4. Department of General Surgery, The First Affiliated Hospital of Medical School, Xian Jiaotong University, Xi'an, 710061, Shaanxi, China. v84239141@hotmail.com. 5. College of Life Science, Shaanxi Normal University, Xi'an, 710119, Shaanxi, China. ychhou@snnu.edu.cn. 6. College of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, 710119, Shaanxi, China. yin_sw@snnu.edu.cn. 7. Division of Gastroenterology and Hepatology, Department of Medicine, School of Medicine, University of Michigan Ann Arbor, Ann Arbor, MI, 48109, USA. thomaswa@med.umich.edu. 8. Department of Gastroenterology, The First Affiliated Hospital of Medical School, Xian Jiaotong University, Xi'an, 710061, China. hesx123@126.com. 9. Department of General Surgery, The First Affiliated Hospital of Medical School, Xian Jiaotong University, Xi'an, 710061, Shaanxi, China. robertlu@mail.xjtu.edu.cn.
Abstract
OBJECTIVE: To develop a peptide probe that could be used for gastric cancer detection via binding to CD44 protein with specificity and affinity. RESULTS: A 12-mer phage peptide library was screened against immobilized CD44 protein. Bound phage counts using ELISA were performed to identify phage clones carrying the most highly selective peptide, which termed RP-1. Immunofluorescence and flow cytometry analysis indicated that the consensus peptide RP-1 could bind to CD44-positive gastric cancer cells with mean fluorescence intensities significantly higher than that of CD44-negative cells. CD44 knockdown led to decreased binding activity of RP-1 to the same cell line. Tissue array technique was used to identify the relationship (r = 0.556) between peptide binding and CD44 detection on gastric cancer tissues. Further, the hyaluronan-binding domain of CD44 was docked with RP-1 using computer modeling/docking approaches, revealing a RP-1/CD44 interaction with geometrical and energy match (-8.6 kcal/mol). CONCLUSIONS: The RP-1 peptide we screened exhibits affinity and specificity to CD44 on cells and has the potential to be used as a candidate probe for gastric cancer cell targeting.
OBJECTIVE: To develop a peptide probe that could be used for gastric cancer detection via binding to CD44 protein with specificity and affinity. RESULTS: A 12-mer phage peptide library was screened against immobilized CD44 protein. Bound phage counts using ELISA were performed to identify phage clones carrying the most highly selective peptide, which termed RP-1. Immunofluorescence and flow cytometry analysis indicated that the consensus peptide RP-1 could bind to CD44-positive gastric cancer cells with mean fluorescence intensities significantly higher than that of CD44-negative cells. CD44 knockdown led to decreased binding activity of RP-1 to the same cell line. Tissue array technique was used to identify the relationship (r = 0.556) between peptide binding and CD44 detection on gastric cancer tissues. Further, the hyaluronan-binding domain of CD44 was docked with RP-1 using computer modeling/docking approaches, revealing a RP-1/CD44 interaction with geometrical and energy match (-8.6 kcal/mol). CONCLUSIONS: The RP-1 peptide we screened exhibits affinity and specificity to CD44 on cells and has the potential to be used as a candidate probe for gastric cancer cell targeting.
Authors: Moraima Morales-Cruz; Yamixa Delgado; Kai Griebenow; Betzaida Castillo; Cindy M Figueroa; Anna M Molina; Anamaris Torres; Melissa Milián Journal: Drug Des Devel Ther Date: 2019-10-30 Impact factor: 4.162
Authors: Dan Zhang; Jin Huang; Weiming Li; Zhiyong Zhang; Meng Zhu; Yun Feng; Yan Zhao; Yarui Li; Shaoying Lu; Shuixiang He Journal: Ann Transl Med Date: 2020-11