Antibody Directed Gene Therapy for Breast Cancer
Abstract
The main objective of this proposal is to develop monoclonal antibodies as cell specific targeting vectors so that they are able to bind and carry DNA into cells. The molecularly engineered antibody targeted DNA to tumor cells, but the expression of the DNA was low. Different strategies have been examined to improve the expression. These strategies have included incorporation of a flu fusion peptide into the antibody/DNA complex, DNA condensation, and the role of viral origins of replication in the transport of naked DNA into the nucleus. The major barrier remained the cytoplasmic membrane. To achieve entry into the cytoplasm, the entire domain containing the flu fusion peptide, HA2, was engineered to contain a DNA binding domain, and expressed by the baculovirus/insect cell system. The fused HA2 protein, upon purification, was shown to have the signal peptide still attached. Thus, the HA2 did not have glycine as its N- terminal amino acid, which is essential for its fusogenic property. Thrombin and factor Xa recognition sites were incorporated into the N-terminus of HA2 to provide a way to get a N-terminus glycine in the HA2.
Document Details
- Document Type
- Technical Report
- Publication Date
- Feb 01, 2001
- Accession Number
- ADA391365
Entities
People
- Peter J. Curtis
Organizations
- Thomas Jefferson University