Blood-brain barrier permeable peptide compositions comprising a vab domain of a camelid single domain heavy chain antibody against an amyloid-beta peptide
Blood-brain barrier permeable peptide compositions that contain variable antigen binding domains from camelid and/or shark heavy-chain only single-domain antibodies are described. The variable antigen binding domains of the peptide compositions bind to therapeutic and diagnostic biomarkers in the central nervous system, such as the amyloid-beta peptide biomarker for Alzheimer's disease. The peptide compositions contain constant domains from human IgG, camelid IgG, and/or shark IgNAR. The peptide compositions include heavy-chain only single-domain antibodies and compositions with one or more variable antigen binding domain bound to one or more constant domains.
1. A composition comprising:
a polypeptide comprising the formula Vab1-L1-X-R3-Y-L2-Vab2,
wherein the Vab1 and Vab2 each independently comprises a variable antigen-binding domain from a camelid single-domain heavy chain antibody, wherein at least one of the variable antigen-binding domains comprises amino acids 1-127 of the amino acid sequence of SEQ ID NO:11 and is capable of binding to an amyloid-beta peptide;
wherein L1 and L2 each comprise a hinge region from a camelid single-domain heavy-chain only antibody;
wherein a nanoparticle comprising polybutylcyanoacrylate and dextran is covalently linked to at least one of L1 and L2; and
wherein X and Y are bifunctional linkers, selected from a maleimido-thiol conjugate and polyethylene glycol;
wherein R3 comprises at least one constant domain of human IgG CH2 or CH3 domain.
2. The composition of claim 1 , wherein the nanoparticle has a molecular weight of about 70 kDa.
3. The composition of claim 1 , wherein at least one of L1 and L2 is covalently conjugated to the label selected from the group consisting of a radioisotope, a fluorophores, a toxin, biotin, digoxigenin, avidin, and streptavidin.
4. The composition of claim 1 , wherein a hinge region corresponds to the amino acid sequence of SEQ ID NO: 13.
5. The composition of claim 1 , wherein at least one of L1 and L2 is covalently conjugated to —NH—C(═NH 2 .HCl)—CH 2 CH 2 CH 2 —S-Maleimide-PEG-CONH—, which in turn is conjugated to a nanoparticle.