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Using Phage Display to Select Peptides Binding to Type 8 Capsular Polysaccharide of Staphylococcus aureus

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2016, Master of Science in Biological Sciences, Youngstown State University, Department of Biological Sciences and Chemistry.
S. aureus aureus (S. aureus) is a major bacterial pathogen causing an array of critical infections. Increasing the problem of S. aureus infections is the fact that forty to sixty percent of S. aureus infections have now become antibiotic resistant, making it difficult to treat. S. aureus is protected by a polysaccharide capsule, which contributes to its virulence. The goal of this project is to identify a peptide that will bind to S. aureus type 8 capsular polysaccharide using phage display technology. Phage display is a technique used to display proteins or peptides on the surface of a phage (a virus that infects a bacterium). If a peptide displayed on the phage surface binds to a molecule of interest, the DNA for that peptide can be sequenced and used to synthesize the pure peptide. In this study, S. aureus capsular carbohydrate was purified and the presence of carbohydrate was confirmed using a red tetrazolium carbohydrate assay and an ELISA using anti-capsular hybridoma antibodies. The sample was determined to be free of contaminants using a Bradford assay for protein, phosphate assay for teichoic acid, and a DNA concentration assay for nucleic acid contamination. Using a phage display library, biopanning was performed to identify peptides that bind to whole bacteria. A series of ELISAs were performed to prove binding of the peptide to type 8 capsular polysaccharide of S. aureus. These ELISAs concluded specific clones 1, 2, 3, and 6 showed the most significant amount of binding to the type 8 capsular polysaccharide of S. aureus. Future studies will include further testing of the specific clones, and DNA sequencing of the most specific clone. The peptide can then be purchased and tested for specificity. The clone DNA may then be transferred to expression vectors and coupled to toxins.
Diana Fagan, PhD (Advisor)
David Asch, PhD (Committee Member)
Gary Walker, PhD (Committee Member)
86 p.

Recommended Citations

Citations

  • Lenkey, N. M. (2016). Using Phage Display to Select Peptides Binding to Type 8 Capsular Polysaccharide of Staphylococcus aureus [Master's thesis, Youngstown State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ysu148611183162124

    APA Style (7th edition)

  • Lenkey, Nina. Using Phage Display to Select Peptides Binding to Type 8 Capsular Polysaccharide of Staphylococcus aureus. 2016. Youngstown State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ysu148611183162124.

    MLA Style (8th edition)

  • Lenkey, Nina. "Using Phage Display to Select Peptides Binding to Type 8 Capsular Polysaccharide of Staphylococcus aureus." Master's thesis, Youngstown State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=ysu148611183162124

    Chicago Manual of Style (17th edition)