Modular functional peptides for delivery of nanoparticles
Inventors
Delehanty, James B. • Medintz, Igor L. • Mattoussi, Hedi M. • Deschamps, Jeffrey R. • Dawson, Glyn • Dawson, Philip E. • Blanco-Canosa, Juan Bautista • Boeneman, Kelly
Assignees
Publication Number
US-8835172-B2
Publication Date
2014-09-16
Expiration Date
2030-10-20
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
A peptide directs nanoparticles (such as quantum dots) to the plasma membrane of mammalian cells. A method of delivery of a nanoparticle to a plasma membrane of a cell includes providing to the cell a nanoparticle attached to a peptide configured to direct the nanoparticle the plasma membrane, and allowing the cell to take up the nanoparticle. The nanoparticle can be a FRET donor to an organic dye.
Core Innovation
Described herein is an invention of modular, multifunctional peptides designed for the delivery of nanoparticles, such as quantum dots, into mammalian cells. These peptides comprise multiple functional domains: a nanoparticle association domain, a spacer domain, an uptake domain, and a vesicle escape domain containing a non-hydrolyzable lipid moiety. The spacer domain functions to reduce steric hindrance between the nanoparticle and the peptide, often comprising a polyproline sequence. When attached to nanoparticles, these peptides mediate the cellular uptake and enable the escape of nanoparticles from endocytic vesicles to the cytosol.
The background of the invention addresses the deficiencies of existing methods for intracellular delivery of nanoparticles. Common methods such as passive delivery, facilitated delivery with endocytosis, and active delivery by physical manipulation each have shortcomings: nonspecific uptake, endosomal sequestration requiring further endosomal disruption which can be toxic, and highly invasive procedures that reduce cell viability. The invention improves intracellular delivery by engineering peptides that not only facilitate uptake but also promote endosomal escape efficiently and with minimal cytotoxicity.
Claims Coverage
The patent claims are centered around methods of labeling and delivering nanoparticles to cells using peptides, specifically involving peptides with modular domains for targeting and delivery functionalities. The claims include methods of labeling plasma membranes and cells, delivering nanoparticles with peptide conjugates, and advantages regarding endocytosis inhibition.
Method for labeling plasma membrane with peptide-nanoparticle conjugates and FRET
Providing to a cell an organic fluorescent dye adapted to partition to the plasma membrane and a nanoparticle attached to a peptide comprising JB858, wherein the nanoparticle acts as a FRET donor to the organic dye; followed by photo-excitation of the nanoparticle to obtain fluorescence from the organic dye in the plasma membrane.
Method for labeling mammalian cells with two nanoparticle-peptide conjugates
Providing to the cell a first nanoparticle attached to a first peptide comprising JB858 to target plasma membrane labeling, and a second nanoparticle attached to a modular peptide comprising a nanoparticle association domain, spacer domain, uptake domain, and a vesicle escape domain with a non-hydrolyzable lipid moiety; where the second peptide induces nanoparticle uptake and delivery to the cytosol.
Method of plasma membrane targeting and uptake of nanoparticles using JB858 peptide
Providing to a cell a nanoparticle attached to a peptide comprising JB858 and allowing cellular uptake, which results in localization to the plasma membrane and partial inhibition of endocytosis.
Collectively, the claims cover novel methods of using modular peptides, particularly those comprising JB858, to direct nanoparticles to plasma membranes, enable cytosolic delivery via endosomal escape domains, and produce functional labeling using FRET mechanisms. They encompass compositions and delivery techniques facilitating selective cellular localization and uptake while inhibiting undesired endocytosis.
Stated Advantages
Efficient delivery of nanoparticles to the cytosol by mediation of cellular uptake and endosomal escape using multifunctional peptides.
Minimal cytotoxicity compared to other delivery methods involving polymers or chemicals.
Stable association of peptide and nanoparticle within cells over multiple days, enabling long-term imaging and labeling.
Capability to specifically target plasma membranes and achieve combinatorial cellular labeling with multiple nanoparticles.
Inhibition of endocytosis by peptide-nanoparticle conjugates allows enhanced targeting to plasma membranes.
Improved photostability of fluorescent dyes through FRET excitation mediated by nanoparticle-peptide conjugates.
Documented Applications
General cellular labeling using nanoparticle-peptide conjugates.
Labeling of specific subcellular compartments, including plasma membrane and cytosol.
Cellular tagging for cell sorting and cataloging purposes.
Delivery of biological cargos such as proteins and drugs to the cytosol.
In vivo imaging applications utilizing biofunctionalized nanoparticles.
FRET-based sensing techniques employing nanoparticles as energy donors to fluorescent dyes localized in the plasma membrane.
Interested in licensing this patent?