Modular platform for targeted therapeutics
A lipidated secondary antibody is disclosed. Particles comprising same are also disclosed.
1 . A lipid nanoparticle comprising a lipidated polypeptide non-covalently attached to an outer surface of the lipid nanoparticle, wherein the lipidated polypeptide is a fusion protein comprising, from N-terminus to C-terminus, an inner membrane lipidation signal peptide from an inner membrane bacterial lipoprotein for anchoring the lipidated polypeptide to the lipid nanoparticle non-covalently and an antibody fragment selected from the group consisting of a Fv, a single chain Fv (scFv), a disulfide-stabilized Fv (dsFv), an Fab, an Fab′, and an F(ab′) 2 that selectively binds an antigen.
2 . The lipid nanoparticle of claim 1 , wherein said antibody fragment comprises a scFv, a variable region of a light chain, or an Fd fragment of a heavy chain.
3 . The lipid nanoparticle of claim 1 , wherein said antibody fragment is a humanized or human antibody fragment.
4 . The lipid nanoparticle of claim 1 , wherein said inner membrane lipidation signal peptide comprises SEQ ID NO: 13.
5 . The lipid nanoparticle of claim 1 , wherein said inner membrane lipidation signal peptide consists of SEQ ID NO: 13.
6 . The lipid nanoparticle of claim 1 , wherein said lipid nanoparticle comprises a charged external surface.
7 . The lipid nanoparticle of claim 1 , wherein said lipid nanoparticle comprises cationic lipids.
8 . The lipid nanoparticle of claim 1 , wherein said lipid nanoparticle is loaded with a diagnostic or a therapeutic agent.
9 . The lipid nanoparticle of claim 8 , wherein said therapeutic agent is a nucleic acid agent.
10 . The lipid nanoparticle of claim 9 , wherein said nucleic acid agent is a DNA, RNA, siRNA, plasmid DNA, short-hairpin RNA, small temporal RNA (stRNA), microRNA (miRNA), RNAmimetic or heterochromatic siRNA.
11 . The lipid nanoparticle of claim 10 , wherein said nucleic acid agent is a siRNA.
12 . A method of producing the lipid nanoparticle of claim 1 , the method comprising incubating a lipidated polypeptide that is a fusion protein comprising, from N-terminus to C-terminus, an inner membrane lipidation signal peptide from an inner membrane bacterial lipoprotein and an antibody fragment selected from the group consisting of a Fv, a single chain Fv (scFv), a disulfide-stabilized Fv (dsFv), an Fab, an Fab′, and an F(ab′) 2 that selectively binds an antigen with a lipid nanoparticle, wherein the inner membrane lipidation signal peptide anchors the lipidated polypeptide fusion protein to the lipid nanoparticle.
13 . The method of claim 12 , wherein said method comprises isolating said lipidated polypeptide from a bacteria into detergent micelles prior to said incubating.
14 . A method of delivering a diagnostic or a therapeutic agent to a subject in need thereof, the method comprising administering to the subject the lipid nanoparticle of claim 8 , thereby delivering the diagnostic or the therapeutic agent to the subject.
15 . The lipid nanoparticle of claim 1 , wherein said inner membrane bacterial lipoprotein is selected from the group consisting of AraH, MglC, MalF, MalG, MalC, MalD, RbsC, RbsC, ArtM, ArtQ, GlnP, ProW, HisM, HisQ, LivH, LivM, LivA, LivE, DppB, DppC, OppB, AmiC, AmiD, BtuC, FhuB, FecC, FecD, FecR, FepD, NikB, NikC, CysT, CysW, UgpA, UgpE, PstA, PstC, PotB, PotC, PotiH, Potl, ModB, NosY, PhnM, LacY, SecY, TolC, DsbB, DsbD, TonB, TatC, CheY, TraB, Exb D, ExbB and Aas.
16 . The lipid nanoparticle of claim 1 , wherein said inner membrane bacterial lipoprotein is E. coli new lipoprotein A (NlpA).
17 . The lipid nanoparticle of claim 1 , wherein said inner membrane bacterial lipoprotein is K. pneumoniae pullulanase.
18 . The lipid nanoparticle of claim 1 , wherein said inner membrane lipidation signal peptide comprises SEQ ID NO: 17.