Production and Characterization of an Immune Phage Display Library to Glioblastoma
The purpose of the study is to develop a collection of binding particles (antibody phage display library), scFV fragments (single chain fragment variable region) that recognize unique sites on antigens present on glioblastoma tumor cells that could be of use for therapeutic and/or diagnostic purposes. These scFV fragments represent the molecular engineered binding portion of antibody molecules that recognize unique epitopes (specific sequences of amino acids) present on glioblastoma antigens (protein product of a mutated normal gene or the product of a gene that is normally developmentally silenced). This proposal represents the initial stage in a research program that has the long-term goals to develop, test, and obtain approval for clinical trials of a cocktail of scFV fragments covalently linked to a substance lethal to the cell (termed immunotoxin) which, on binding specific epitopes, would kill the glioblastoma cells.
The graduate research candidate will use molecular techniques to produce cDNA from spleens of immunized mice. Then use multiplex PCR to amplify hyper veriable regions from coding regions for both heavy and light chains. The PCR products will be cloned into phagemid expression constructs and then electroporated into E. coli for expression of the bactrophage now containing the cloned insert. This colection of bacteriophage particles will represent the phage library which will be screened for expression of surface sites that will bind to glioblastoma antigen. Immune absorption, Enzyme linked immunosorbent assays (ELISAs) and western blots will be used to identify and expand phage particles that bind to specific glioblastoma antigens.In some cases additional mice immunizations may be conducted.
1. Expertise in RNA isolation and agarose gel analysis as well as other molecular biology techniques.
2. Experience with western blot testing. Including preparation of antigen strips as well as the actual immunlogical testing.
3. Good aseptic technique and experience working under a biosafety hood.
4. Completion of relevent University of Michigan courses required for carr and use of laboratory mice.
The work schedule will be varriable depending on the particular experiment(s) that are being conducted. In some instances it may be daily for 1.5 hrs occuring five to seven days a week in other instances one experiment may involve a six hour time block on one day, and in others it may be 12 hours spread over 2-3 days. Timing to execute a particular experiment within a day is flexiable.
This project is a blend of molecular biology, theoritical immunology, and applied immunology. The ability to use modern techniques to iolate RNA is an absolute requirement. It also may involve working with mice and previous experience university training is desired.
- Job Opening ID
- Winter 2021 Only
- Work could be done by someone not coming to campus (e.g., online or non-local student)
- What majors can apply?
- Biology (MS)
- Faculty Name
- Jerry Sanders