Humanized docking system for assembly of targeting drug delivery complexes.
Targeted drug delivery requires 'loading' drugs onto targeting proteins. Traditional technologies for loading drugs rely on chemical conjugation of drugs or drug carriers to targeting proteins. An alternative approach might rely on assembly of targeting complexes using a docking system that includes two components: a 'docking' tag fused to a targeting protein, and a 'payload' module containing an adapter protein for non-covalent binding to the docking tag. We describe here a fully humanized adapter/docking tag system based on non-covalent interaction between two fragments of human pancreatic RNase I. A 15 amino acid long N-terminal fragment of RNase I designed to serve as a docking tag, was fused to the N-terminus of human vascular endothelial growth factor that served as a targeting protein. An 18-125 and an 18-127 amino acid long fragments of RNase I were engineered, expressed and refolded into active conformations to serve as adapter proteins. Interactions between the targeting and adapter proteins were characterized using enzymatic analysis and surface plasmon resonance. Targeting DNA delivery complexes were assembled, characterized by dynamic light scattering, and found to be very effective in receptor-mediated DNA delivery.
Backer
Marina V
MV
SibTech, Inc, 705 North Mountain Road, Newington, CT 06111, USA. mbacker@sibtech.com
Gaynutdinov
Timur I
TI
Gorshkova
Inna I
II
Crouch
Robert J
RJ
Hu
Tao
T
Aloise
Renee
R
Arab
Mohamed
M
Przekop
Kristen
K
Backer
Joseph M
JM
D002460:Cell Line; D016503:Drug Delivery Systems / Q000379:methods*; D006801:Humans; D008565:Membrane Proteins / Q000008:administration & dosage* / Q000493:pharmacokinetics; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D008565:Membrane Proteins; C035026:SRPRA protein, human; D042461:Vascular Endothelial Growth Factor A; D012259:Ribonuclease, Pancreatic