Research Project
3488380
The goal of the project is the development of a commercially feasible system for the production of human granulocyte-macrophage colony stimulating factor (GmCSF) for use as a therapeutic agent in the treatment of patients whose immune system has been suppressed as a result of treatment with radiation therapy or chemotherapeutic agents. In Phase I the DNA coding sequence for GmCSF will be cloned and expressed in a mammalian cell culture system. The GmCSF cDNA will be obtained by screening cDNA libraries with oligonucleotide probes designed from the published sequence of murine GmCSF or by screening a phage expression cDNA library with radiolabeled GmCSF antibody. Oligonucleotides or nick translated cDNA probes will be used to obtain the genomic GmCSF DNA. From the cDNA and/or genomic DNA a coding sequence will be assembled. This sequence will be expressed as biologically active GmCSF in tissue culture cells. Expression of biologically active GmCSF will provide information for determining the feasibility of obtaining production level expression in Phase II. The anticipated use of GmCSF will be to enhance the proliferation of granulocytes and macrophage in immune suppressed patients, thereby providing an innovative approach in combination with conventional cell replacement or antibiotic therapy.
