title: Functionalized linear poly(amidoamine)s are efficient vectors for intracellular protein delivery.
abstract: An effective intracellular protein delivery system was developed based on functionalized linear poly(amidoamine)s (PAAs) that form self-assembled cationic nanocomplexes with oppositely charged proteins. Three differently functionalized PAAs were synthesized, two of these having repetitive disulfide bonds in the main chain, by Michael-type polyaddition of 4-amino-1-butanol (ABOL) to cystamine bisacrylamide (CBA), histamine (HIS) to CBA, and ABOL to bis(acryloyl)piperazine (BAP). These water-soluble PAAs efficiently condense β-galactosidase by self-assembly into nanoscaled and positively-charged complexes. Stable under neutral extracellular conditions, the disulfide-containing nanocomplexes rapidly destabilized in a reductive intracellular environment. Cell-internalization and cytotoxicity experiments showed that the PAA-based nanocomplexes were essentially non-toxic. β-Galactosidase was successfully internalized into cells, with up to 94% of the cells showing β-galactosidase activity, whereas the enzyme alone was not taken up by the cells. The results indicate that these poly(amidoamine)s have excellent properties as highly potent and non-toxic intracellular protein carriers, which should create opportunities for novel applications in protein delivery.
authors.affiliation: Department of Biomedical Chemistry, MIRA Institute for Biomedical Technology and Technical Medicine, Faculty of Science and Technology, University of Twente, P.O. Box 217, 7500 AE Enschede, The Netherlands.
authors.forename: Grégory
authors.initials: G
authors.lastname: Coué
authors.forename: Johan F J
authors.initials: JF
authors.lastname: Engbersen
mesh_terms: D016503:Drug Delivery Systems; D006863:Hydrogen-Ion Concentration; D053758:Nanoparticles; D011073:Polyamines / Q000008:administration & dosage / Q000737:chemistry*; D011506:Proteins / Q000008:administration & dosage*; D001616:beta-Galactosidase / Q000008:administration & dosage / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology
chemical_list: C531249:Poly(amidoamine); D011073:Polyamines; D011506:Proteins; D001616:beta-Galactosidase