WHDL - 00011612
WHDL - 00011612
Glioblastomas currently make up 45.2% of all malignant brain tumors and have an extremely poor survival rate due to lack of effective treatment options. The following study examines the possible application of ADEPT (Antibody Directed Enzyme Prodrug Therapy) as a treatment for glioblastomas. This method would allow for on-site activation of an inactive, masked pro-methotrexate molecule (FDA approved) by a mutated Carboxypeptidase A (mCPA) enzyme. Designing for selective activation of the methotrexate at the source of the tumor allows for directed cancer treatment while substantially reducing side effects. Under the supervision of Mike Dorrell, PhD and Heidi Woelbern, PhD, a point mutation for the active site of CPA was designed to generate an expression construct with the mutated version of CPA. This point mutation expands the active site allowing for the extra large amino acid-like ‘mask’ of pro-methotrexate to be cleaved, and the methotrexate drug thus activated. This point mutation of a natural human enzyme prevents rejection and is specific enough that endogenous wild type enzymes will not activate the drug. HEK 293 cells were transfected with the construct to produce the enzyme. In this study, transfection protocols were optimized and purification processes were tested and optimized for protein production and identification.
Arete: The PLNU Honors Journal
This collection includes projects completed by Point Loma Nazarene University undergraduate honors scholars. These projects are completed under the supervision of a faculty advisor or committee and cover a wide variety of disciplines.