Insulin-like growth factor 1 receptor-specific antibodies and uses thereof
The blood-brain barrier (BBB) prevents transport of molecules larger than 500 Daltons from blood to brain. Receptor-mediated transcytosis (RMT) facilitates transport across the BBB of specific molecules that bind receptors on brain endothelial cells that form the BBB. An insulin-like growth factor 1 receptor (IGF1R)-binding antibody or fragment thereof is identified that transmigrates the BBB by RMT. The antibody or fragment is used to deliver a cargo molecule across the BBB, wherein the cargo molecule may be a therapeutic or detectable agent. The antibody is a camelid VHH, prepared by immunizing a llama with a 933-amino acid IGF1R polypeptide. Humanized forms of the camelid VHH are also generated.
1. An isolated or purified single domain antibody or antigen-binding fragment thereof, comprising
a complementarity determining region (CDR) 1 sequence of EYPSNFYA (SEQ ID NO:1);
a CDR2 sequence of VSRDGLTT (SEQ ID NO:2); and
a CDR3 sequence of AIVITGVWNKVDVNSRSYHY (SEQ ID NO:3),
wherein the single domain antibody or antigen-binding fragment thereof is specific for Insulin-Like Growth Factor 1 Receptor (IGF1R).
2. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 , comprising the sequence
X 1 VX 2 LX 3 ESGGGLVQX 4 GGSLRLSCX 5 ASEYPSNFYAMSWX 6 RQAPGKX 7 X 8 EX 9 V X 10 GVSRDGLTTLYADSVKGRFTX 11 SRDNX 12 KNTX 13 X 14 LQMNSX 15 X 16 AEDTAV YYCAIVITGVWNKVDVNSRSYHYWGQGTX 17 VTVSS (SEQ ID NO:4), where X 1 is E or Q; X 2 is K or Q; X 3 is V or E; X 4 is A or P; X 5 is V or A; X 6 is F or V; X 7 is E or G; X 8 is R or L; X 9 is F or W; X 10 is A or S; X 11 is M or I; X 12 is A or S; X 13 is V or L; X 14 is D or Y; X 15 is V or L; X 16 is K or R; and X 17 is Q or L.
3. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 , comprising a sequence selected from the group consisting of:
(SEQ ID NO: 5)
QVKLEESGGGLVQAGGSLRLSCVASEYPSNFYAMSWFRQAPGKEREFVAG
VSRDGLTTLYADSVKGRFTMSRDNAKNTVDLQMNSVKAEDTAVYYCAIVI
TGVWNKVDVNSRSYHYWGQGTQVTVSS;
(SEQ ID NO: 6)
EVQLVESGGGLVQPGGSLRLSCAASEYPSNFYAMSWVRQAPGKGLEWVSG
VSRDGLTTLYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAIVI
TGVWNKVDVNSRSYHYWGQGTLVTVSS;
(SEQ ID NO: 7)
QVQLVESGGGLVQPGGSLRLSCAASEYPSNFYAMSWVRQAPGKGLEWVAG
VSRDGLTTLYADSVKGRFTMSRDNSKNTVYLQMNSLRAEDTAVYYCAIVI
TGVWNKVDVNSRSYHYWGQGTLVTVSS;
(SEQ ID NO: 8)
QVQLVESGGGLVQPGGSLRLSCAASEYPSNFYAMSWFRQAPGKGLEFVAG
VSRDGLTTLYADSVKGRFTMSRDNSKNTVYLQMNSLRAEDTAVYYCAIVI
TGVWNKVDVNSRSYHYWGQGTLVTVSS;
(SEQ ID NO: 9)
QVQLVESGGGLVQPGGSLRLSCAASEYPSNFYAMSWFRQAPGKEREFVAG
VSRDGLTTLYADSVKGRFTMSRDNSKNTVYLQMNSLRAEDTAVYYCAIVI
TGVWNKVDVNSRSYHYWGQGTLVTVSS;
and
(SEQ ID NO: 10)
EVQLVESGGGLVQPGGSLRLSCAASEYPSNFYAMSWFRQAPGKEREFVSG
VSRDGLTTLYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAIVI
TGVWNKVDVNSRSYHYWGQGTLVTVSS.
4. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 , wherein the single domain antibody is of camelid origin.
5. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 , wherein the single domain antibody or antigen-binding fragment thereof is in a multivalent display format.
6. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 5 , wherein the single domain antibody or antigen-binding fragment thereof is linked to a Fc fragment.
7. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 , wherein the single domain antibody or antigen-binding fragment thereof is immobilized onto a surface.
8. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 , wherein the single domain antibody or antigen-binding fragment thereof is linked to a cargo molecule.
9. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 8 , wherein the cargo molecule has a molecular weight in the range of about 1 kD to about 200 kDa.
10. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 8 , wherein the cargo molecule is a detectable agent, a therapeutic, a peptide, a growth factor, a cytokine, a receptor trap, a chemical compound, a carbohydrate moiety, an enzyme, an antibody or fragment thereof, a DNA-based molecule, a viral vector, or a cytotoxic agent; one or more liposomes or nanocarriers loaded with a detectable agent, a therapeutic, a peptide, an enzyme, an antibody or fragment thereof, a DNA-based molecule, a viral vector, or a cytotoxic agent; or one or more nanoparticle, nanowire, nanotube, or quantum dots.
11. A composition comprising one or more than one isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 and a pharmaceutically-acceptable carrier, diluent, or excipient.