GPCR binding proteins and synthesis thereof

Patent metadata

Field	Value
Publication number	US-11407837-B2
Application number	US-201816128372-A
Country	US
Kind code	B2
Filing date	Sep 11, 2018
Priority date	Sep 11, 2017
Publication date	Aug 9, 2022
Grant date	Aug 9, 2022

Abstract

Provided herein are methods and compositions relating to G protein-coupled receptor (GPCR) libraries having nucleic acids encoding for a scaffold comprising a GPCR binding domain. Libraries described herein include variegated libraries comprising nucleic acids each encoding for a predetermined variant of at least one predetermined reference nucleic acid sequence. Further described herein are protein libraries generated when the nucleic acid libraries are translated. Further described herein are cell libraries expressing variegated nucleic acid libraries described herein.

First claim

What is claimed is:

1. A nucleic acid library comprising a plurality of nucleic acids, wherein each nucleic acid comprises:
a) a nucleic acid sequence comprising a length of about 80 to about 230 base pairs that encodes for a CDR-H3 loop, wherein the CDR-H3 loop comprises a sequence of any one of SEQ ID NOs: 2420 to 2436; and
b) a nucleic acid sequence encoding a variable domain of light chain (VL), wherein the nucleic acid sequence comprises at least about 90% sequence identity to any one of SEQ ID NOs: 94, 96, 98, 283, 301, 321, 416, 432, 459, 460, 616, 618, 647, 652, 660, 666, 677, 745, 755, 1627, 1883, and 2043,

wherein each of the nucleic acids encodes for a sequence that when translated encodes for an immunoglobulin scaffold that comprises a GLP1R binding domain, and wherein the GLP1R binding domain is a ligand for GLP1R.

2. The nucleic acid library of claim 1 , wherein the immunoglobulin scaffold further comprises one or more domains selected from variable domain of heavy chain (VH), constant domain of light chain (CL), and constant domain of heavy chain (CH).

3. The nucleic acid library of claim 2 , wherein a length of the VH domain is about 280 to about 300 base pairs.

4. The nucleic acid library of claim 1 , wherein a length of the VL domain is about 90 to about 120 amino acids.

5. The nucleic acid library of claim 1 , wherein a length of the VL domain is about 300 to about 350 base pairs.

6. The nucleic acid library of claim 1 , wherein the library comprises at least 10 5 non-identical nucleic acids.

7. The nucleic acid library of claim 1 , wherein the immunoglobulin scaffold comprises a single immunoglobulin domain.

8. The nucleic acid library of claim 1 , wherein the immunoglobulin scaffold comprises a peptide of at most 100 amino acids.

9. The nucleic acid library of claim 1 , wherein the nucleic acid sequence encoding a variable domain of light chain (VL) comprises a sequence having at least about 90% sequence identity to SEQ ID NOs: 94, 96, and 98.

10. The nucleic acid library of claim 1 , wherein the nucleic acid sequence encoding a variable domain of light chain (VL) comprises a sequence having at least about 90% sequence identity to any one of SEQ ID NOs: 616, 618, 647, 652, 660, 666, 677, 745, and 755.

Assignees

• Twist Bioscience Corp

Inventors

• Glanville Jacob

Classifications

• C07K16/2869 Primary

  against hormone receptors (for antibodies against neuromediator receptors C07K16/286) · CPC title

• C07K16/46

  Hybrid immunoglobulins (hybrids of an immunoglobulin with a peptide not being an immunoglobulin C07K19/00) · CPC title

• C12N15/1044

  Preparation or screening of libraries displayed on scaffold proteins · CPC title

• C07K16/005 Primary

  constructed by phage libraries · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title