Production of monoclonal antibodies against pathogenic Staphylococcus aureus by phage display | |
| Author | Khin Ma Ma Htwe |
| Call Number | AIT Thesis no.FB-05-05 |
| Subject(s) | Monocronal antibodies Bacteriophages |
| Note | A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science, School of Environment, Resources and Development |
| Publisher | Asian Institute of Technology |
| Series Statement | Thesis ; no. FB-05-05 |
| Abstract | The main aim of this study was to produce monoclonal antibodies (MAbs) against foodborne pathogens by phage display for their potential use in therapeutic and diagnostic applications. MAbs were isolated from human nave scFv phage library synthesized from complementary DNA (cDNA) derived from B lymphocytes of 20 healthy donors. The scFv library construction involved amplifying VH and VL with degenerate primers, constructing V„ and VL libraries in pGEM-T Easy, assembling scFv by SOE-PCR, constructing scFv library in pCRII-TOPO, subcloning scFv library into pCANTAB 5E, and generating scFv library in M 13 phage by phage rescue with M I 3KO7. The scFv phage library was screened against 25 common food-borne pathogens by cellular phage ELISA (CP-ELISA), followed by biopanning against selected whole-cell pathogen(s) to identify specific MAbs. The size of scFv library obtained in pCRII-TOPO and pCANTAB 5E was 4.3 x 104 and 1.9 x 104, respectively. The scFv phage library was reactive to 14 pathogens with highest reactivity to S. aureus. Biopanning and CP-ELISA identified 2 S. aureus-specific monoclones (26 and 35), which had moderate cross reactivity to other pathogens |
| Year | 2005 |
| Corresponding Series Added Entry | Asian Institute of Technology. Thesis ; no. FB-05-05 |
| Type | Thesis |
| School | School of Environment, Resources, and Development (SERD) |
| Department | Department of Food, Agriculture and Natural Resources (Former title: Department of Food Agriculture, and BioResources (DFAB)) |
| Academic Program/FoS | Food Engineering and Bioprocess Technology (FB) |
| Chairperson(s) | Rakshit, Sudip Kumar; |
| Examination Committee(s) | Wanpen Chaicumpa ;Ba-Theinm William ;Athapol Noomhorm ;Jindal, Vinod K.; |
| Scholarship Donor(s) | Asian Institute of Technology fellowship; |
| Degree | Thesis (M.Sc.) - Asian Institute of Technology, 2005 |