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Phage random peptide library screening serum tumor markers in nasopharyngeal experimental study

Author: LiuXinQiang
Tutor: HeHaiPing
School: Guangxi Medical University
Course: Oncology
Keywords: Phage peptide library Nasopharyngeal Tumor markers
CLC: R739.63
Type: Master's thesis
Year: 2011
Downloads: 26
Quote: 0
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Abstract

Background and Purpose NPC (Nasopharyngeal Carcinoma, NPC) is a polygenic disease (polygenetic diseases). Currently the diagnosis of nasopharyngeal carcinoma mainly by tissue biopsy, because early symptoms, the patient's five year survival rate is about 50%. However, cancer patients has been shown in vivo to produce the corresponding tumor markers, we can detect these tumor markers for the early diagnosis of NPC. By molecular biology and immunology methods to find those sensitivity, specificity, relatively high tumor marker for early diagnosis of NPC patients to provide new ideas. Phage display technology (phage display technique, PDT) is looking for a specific binding with the target protein polypeptides or proteins relations potent biological tool, its biggest advantage is that can be presented directly to the phenotype and the genotype linked demonstrated out of the foreign protein can be maintained relatively independent spatial structure and biological activity, and use its affinity for the ligand specificity protein or polypeptide of interest screened out, and then sequenced to obtain the corresponding structural and functional information. We used phage display peptide library screening nasopharyngeal carcinoma patients, Amoy selected patients with nasopharyngeal carcinoma tumor antigen-specific expression of the hope for the diagnosis and prognosis of patients with nasopharyngeal carcinoma provide new markers. Method 1. Were used normal human serum and serum of patients with nasopharyngeal carcinoma coated microtiter plate, through the reduction of the screening method, the pre-adsorbed phage with normal serum, and then unbound phage serum of patients with NPC again adsorption, then get nasopharyngeal carcinoma patients with a combination of phage amplification. Repeat screening times. (2) pick a single plaque with screening detected by ELISA in serum of patients with nasopharyngeal carcinoma serum binding situation, choose significantly different phages, and then expand the content of serum samples for testing, and ultimately get to work with patients with nasopharyngeal carcinoma a single serum specific binding phage clones. 3 Extraction and purification of single phage clone DNA, using primers -96 Ⅲ DNA sequencing, according to the peptide and amino acid insertion point into the genetic code table to obtain a short peptide sequence. Results 1 After three screening, pick a single phage obtained after repeatedly identified two specific phage clones better; 2. Were sequenced arrangement for these two short peptide sequences; 3. These two phage clones were identified IgA-ELISA VCA-positive nasopharyngeal carcinoma serum and serum IgA-VCA-negative nasopharyngeal no significant difference; 4 which two phage clones were detected by ELISA in serum nasopharyngeal carcinoma, liver cancer sera sera showed that ovarian cancer patients with nasopharyngeal carcinoma has a specific binding. Conclusions In this study, phage random peptide library screening with nasopharyngeal carcinoma-associated antigen-specific binding peptides for the early diagnosis of nasopharyngeal carcinoma, joint detection and immunotherapy of candidate markers.

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CLC: > Medicine, health > Oncology > Department of Otolaryngology tumor > Pharyngeal tumors
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