METHOD FOR PRODUCING AN EXAMINATION REAGENT AND KIT FOR ANALYSING A T-CELL FREQUENCY

• US 20190331681A1
• Filed: 07/01/2019
• Published: 10/31/2019
• Est. Priority Date: 01/04/2012
• Status: Active Grant

• Current Assignee: Constructor University Bremen Ggmbh
• Original Assignee: Jacobs University Bremen Ggmbh
• Inventors: SPRINGER, SEBASTIAN HARTMUT, SAINI, SUNIL KUMAR
• Granted Patent: US 11,698,375 B2
• Time in Patent Office: Days
• CPC Class Codes: C07K 14/70539 MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: G01N 2333/70539 MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
• CPC Class Codes: G01N 33/56977 HLA or MHC typing

Abstract

A method for producing an examination reagent includes adding a helper ligand to a receptor protein which is unfolded in an initial solution so as to provide a pre-solution of the examination reagent with a receptor protein which is folded. The receptor protein which is folded comprises a bonded helper ligand which can be exchanged with an examination peptide.

Claims

1. A method for producing an examination reagent, the method comprising:

adding a helper ligand to a receptor protein which is unfolded in an initial solution so as to provide a pre-solution of the examination reagent with a receptor protein which is folded,

wherein, the receptor protein which is folded comprises a bonded helper ligand which can be exchanged with an examination peptide.

2. The method as recited in claim 1, wherein the receptor protein which is unfolded is an unfolded MHC class I protein.

3. The method as recited in claim 1, further comprising:

adding a ligand exchanger/ligand releaser to the receptor protein which is folded.

4. The method as recited in claim 3, wherein the ligand exchanger/ligand releaser has an affinity for the receptor protein which is less than an affinity for the helper ligand.

5. The method as recited in claim 3, wherein the ligand exchanger/ligand releaser is an amino acid or a peptide comprising 2, 3, 4, 5, 6 or 7 amino acids.

6. The method as recited in claim 1, wherein at least one of:

the helper ligand has an affinity for the receptor protein which is less than an affinity of the examination peptide, and

the helper ligand is a peptide consisting of 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 amino acids.

7. The method as recited in claim 1, wherein the receptor protein which is unfolded is produced in an expression system.

8. The method as recited in claim 1, further comprising:

attaching a signaling element to the receptor protein which is unfolded or to the receptor protein which is folded,

wherein, the signaling element is a fluorescent dye.

9. The method as recited in claim 1, further comprising:

attaching an anchor element to the receptor protein which is unfolded or to the receptor protein which is folded,

wherein, the anchor element is a biotin molecule.

10. The method as recited in claim 1, further comprising:

providing a plurality of receptor proteins each of which are folded; and

assembling the plurality of receptor proteins each of which are folded to form a multimer,

wherein, at least one of the receptor proteins which is folded comprises the helper ligand.

11. The method as recited in claim 10, further comprising:

attaching a multimer signaling element to the multimer,

wherein, the multimer signaling element is a fluorescent dye.