Methods and compositions related to soluble monoclonal variable lymphocyte receptors of defined antigen specificity

Patent metadata

Field	Value
Publication number	US-11945857-B2
Application number	US-202017126396-A
Country	US
Kind code	B2
Filing date	Dec 18, 2020
Priority date	Aug 2, 2006
Publication date	Apr 2, 2024
Grant date	Apr 2, 2024

Abstract

Disclosed are compositions and methods related to variable lymphocyte receptors (VLRs). More particularly, disclosed are a variety of antigen specific polypeptides, including soluble, monoclonal, and multivalent forms, as well as methods of using the polypeptides, antibodies that bind the antigen specific polypeptides, and nucleic acids, vectors and expression systems that encode the polypeptides. Antigen specific polypeptides that selectively bind pathogens, like anthrax, and carbohydrates, like blood group determinants, are specifically disclosed.

First claim

What is claimed is:

1. A cell culture, wherein the culture medium comprises a plurality of multimers comprising a plurality of soluble, monoclonal antigen specific polypeptides, wherein each antigen specific polypeptide comprises an N-terminal leucine rich repeat (LRRNT), one or more leucine rich repeats (LRRs), a C-terminal leucine rich repeat (LRRCT), and a connecting peptide, wherein the connecting peptide comprises an alpha helix.

2. The cell culture of claim 1 , wherein the cells comprise a cDNA encoding an antigen specific polypeptide, wherein the antigen specific polypeptide comprises an N-terminal leucine rich repeat (LRRNT), one or more leucine rich repeats (LRRs), a C-terminal leucine rich repeat (LRRCT), and a connecting peptide, wherein the connecting peptide comprises an alpha helix.

3. The cell culture of claim 2 , wherein the cDNA is stably integrated into the genome of the cells.

4. The cell culture of claim 2 , wherein the antigen specific polypeptide binds a target protein, a target carbohydrate or a target pathogen.

5. The cell culture of claim 1 , wherein each multimer comprises up to ten antigen specific polypeptides.

6. The cell culture of claim 1 , wherein each multimer is a multivalent multimer that binds to more than one target.

7. The cell culture of claim 1 , wherein the antigen specific polypeptide is an affinity matured polypeptide.

8. The cell culture of claim 1 , wherein the cell culture is a mammalian cell culture.

9. A method of making the cell culture of claim 1 , comprising transfecting a cultured cell with a cDNA clone encoding an antigen specific polypeptide, wherein the antigen specific polypeptide comprises an N-terminal leucine rich repeat (LRRNT), one or more leucine rich repeats (LRRs), a C-terminal leucine rich repeat (LRRCT), and a connecting peptide, wherein the connecting peptide comprises an alpha helix.

10. The method of claim 9 , wherein the antigen specific polypeptide is an affinity matured polypeptide.

11. The method of claim 9 , further comprising: (a) screening the culture medium for an ability to bind a target antigen; (b) optionally, isolating the plurality of multimers from the culture medium; and (c) modifying the antigen specific polypeptide that binds a target antigen by affinity maturation.

12. The method of claim 11 , wherein affinity maturation comprises phage display, yeast display, bacterial display or ribosome display.

13. The method of claim 9 , further comprising isolating the plurality of multimers from the culture medium.

14. The method of claim 10 , further comprising isolating the plurality of multimers from the culture medium.

15. The method of claim 9 , wherein the antigen specific polypeptides bind a target protein, a target carbohydrate or a target pathogen.

16. The method of claim 9 , wherein each multimer comprises up to ten antigen specific polypeptides.

17. The method of claim 9 , wherein each multimer is a multivalent multimer that binds to more than one target.

18. The method of claim 9 , wherein the cell culture is a mammalian cell culture.

Assignees

• Uab Res Found

Inventors

• Cooper Max D
• Herrin Brantley R
• Alder Matthew N

Classifications

• C07K16/1145

  Env proteins, e.g. gp41, gp110/120, gp160, V3, principal neutralising domain [PND] or CD4-binding site · CPC title

• C07K16/108

  Orthomyxoviridae (F), e.g. influenza virus · CPC title

• C07K16/1278 Primary

  Bacillus (G) · CPC title

• C07K16/1018

  Chemistry & Metallurgy · mapped topic

• C07K16/1063

  Chemistry & Metallurgy · mapped topic