Nuclear delivery of p53 C-terminal peptides into cancer cells using scFv fragments of a monoclonal antibody that penetrates living cells.
scFv fragments of a monoclonal antibody that penetrates living cells and localizes in nuclei were designed as fusion proteins with C-terminal p53 peptides and tested for restoring p53 function in p53 mutant cancer cells. scFv fragments transported a 30-mer C-terminal peptide of p53 into cancer cells and induced cellular cytotoxicity in contrast to scFv fragments alone and other scFv-p53 fusion peptides. Cellular toxicity was not observed with scFv fragments containing a single mutation in VH that prevented antibody penetration. Our results demonstrate the potential efficacy of antibody scFv fragments as a nuclear delivery system in cancer cells.
Weisbart
Richard H
RH
Department of Medicine, Division of Rheumatology, Veterans Affairs Greater Los Angeles Health Care System, Sepulveda, CA 91343, USA. rweisbar@ucla.edu
Miller
Carl W
CW
Chan
Grace
G
Wakelin
Rika
R
Ferreri
Kevin
K
Koeffler
H Phillip
HP
D021581:Active Transport, Cell Nucleus; D000230:Adenocarcinoma / Q000473:pathology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage; D017209:Apoptosis / Q000187:drug effects*; D001859:Bone Neoplasms / Q000473:pathology; D016466:CHO Cells; D019556:COS Cells; D002463:Cell Membrane Permeability; D002467:Cell Nucleus / Q000378:metabolism*; D002522:Chlorocebus aethiops; D015179:Colorectal Neoplasms / Q000473:pathology; D006224:Cricetinae; D016503:Drug Delivery Systems; D017930:Genes, Reporter; D006801:Humans; D018796:Immunoconjugates / Q000008:administration & dosage* / Q000494:pharmacology / Q000633:toxicity; D007135:Immunoglobulin Variable Region / Q000008:administration & dosage* / Q000494:pharmacology; D051379:Mice; D019463:Mice, Inbred MRL lpr; D012516:Osteosarcoma / Q000473:pathology; D010446:Peptide Fragments / Q000008:administration & dosage* / Q000494:pharmacology / Q000633:toxicity; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000494:pharmacology; D015533:Transcriptional Activation / Q000187:drug effects*; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000008:administration & dosage* / Q000737:chemistry / Q000502:physiology / Q000633:toxicity
D000911:Antibodies, Monoclonal; D018796:Immunoconjugates; D007135:Immunoglobulin Variable Region; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D016159:Tumor Suppressor Protein p53