CPC C07K 16/2881 (2013.01) [C07K 16/005 (2013.01); C12N 15/1058 (2013.01); C40B 20/00 (2013.01); C40B 30/04 (2013.01); C40B 50/00 (2013.01); C07K 2317/35 (2013.01); C07K 2317/524 (2013.01); C07K 2317/526 (2013.01); C07K 2317/55 (2013.01); C12N 2320/10 (2013.01)] | 17 Claims |
1. A method of generating one or more variant polypeptides that specifically bind transferrin receptor protein and transport across the blood brain barrier, comprising:
(a) identifying a contiguous surface exposed register of about 7 to 15 amino acids of a native polypeptide for mutation;
(b) generating one or more polynucleotides comprising a nucleotide sequence that encodes for a library of sequence variants of the native polypeptide, wherein the nucleotide sequence encodes for one or more mutations in the identified register of the native polypeptide;
(c) expressing the one or more polynucleotides to produce the library of sequence variants;
(d) screening the library of sequence variants to identify one or more variant polypeptides that specifically bind transferrin receptor protein; and
(e) isolating the variant polypeptides,
wherein the identified register of the native polypeptide does not bind transferrin receptor protein; and the isolated variant polypeptides exhibit substantially improved brain uptake compared to the native polypeptide.
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