US 11,912,778 B2
Methods of engineering transferrin receptor binding polypeptides
Xiaocheng Chen, South San Francisco, CA (US); Mark S. Dennis, South San Francisco, CA (US); Mihalis Kariolis, South San Francisco, CA (US); Adam P. Silverman, South San Francisco, CA (US); Ankita Srivastava, South San Francisco, CA (US); Ryan J. Watts, South San Francisco, CA (US); Robert C. Wells, South San Francisco, CA (US); and Joy Yu Zuchero, South San Francisco, CA (US)
Assigned to Denali Therapeutics Inc., South San Francisco, CA (US)
Filed by Denali Therapeutics Inc., South San Francisco, CA (US)
Filed on Aug. 15, 2022, as Appl. No. 17/888,406.
Application 17/888,406 is a continuation of application No. 16/543,332, filed on Aug. 16, 2019, granted, now 11,795,232.
Application 16/543,332 is a continuation of application No. PCT/US2018/018371, filed on Feb. 15, 2018.
Claims priority of provisional application 62/460,692, filed on Feb. 17, 2017.
Claims priority of provisional application 62/543,658, filed on Aug. 10, 2017.
Claims priority of provisional application 62/583,314, filed on Nov. 8, 2017.
Prior Publication US 2023/0235072 A1, Jul. 27, 2023
Int. Cl. C40B 30/04 (2006.01); C07K 16/28 (2006.01); C07K 16/00 (2006.01); C12N 15/10 (2006.01); C40B 20/00 (2006.01); C40B 50/00 (2006.01)
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
OG exemplary drawing
 
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.