Receptor binding ligands, their use in the detection of cells with biological interest

Patent metadata

Field	Value
Publication number	US-9791435-B2
Application number	US-201013143657-A
Country	US
Kind code	B2
Filing date	Jan 8, 2010
Priority date	Jan 9, 2009
Publication date	Oct 17, 2017
Grant date	Oct 17, 2017

Abstract

A method for the identification and quantification of the expression of membrane receptors present on the surface of target cells using at least two soluble receptor binding ligands derived from the soluble part of the glycoprotein of an enveloped virus that interacts with a cellular cognate receptor, the receptor binding ligand containing a part or the totality of one of the receptor binding domains (RBD) of the glycoprotein, and the soluble receptor binding ligand being liable to interact with the at least one membrane receptor of a target cell, for the identification and quantification of the expression of membrane receptors present on the surface of target cells, the identification and quantification taking place at a given time or during a given time interval, and allowing the determination of a physiological state of the target cell.

First claim

The invention claimed is:

1. A method for determining a physiological state of target cells taking place at a given time or during a given time interval, comprising:
contacting the target cells with at least two soluble receptor binding ligands comprising the totality of the receptor binding domains (RBD) of at least two glycoproteins of at least two enveloped viruses that interacts with a cellular cognate receptor,
each of said at least two receptor binding ligands having a tag, detectable molecule or label coupled thereto, each of said at least two soluble receptor binding ligands containing the totality of one of the RBD of said glycoprotein, wherein said RBD does not comprise a transmembrane domain; and,
wherein complexes are formed between each of said at least two soluble receptor binding ligands and each of at least two distinct membrane receptors, and the complexes are detectable by the tag, the detectable molecule or the label coupled to each of said at least two receptor binding ligands,
wherein all receptor binding ligands contacting the cells contain the totality of RBD of said glycoprotein, and said all receptor binding ligands do not comprise a transmembrane domain, and
wherein said at least two glycoproteins belong to viruses selected from the group consisting of: Amphotropic MLV (ampho, SEQ ID NO:1), Gibbon Ape Leukemia virus (GALV, SEQ ID NO:2), Feline endogenous virus (RD114, SEQ ID NO:3), vesicular stomatitis virus (VSV, SEQ ID NO:4), Xenotropic Murine Leukaemia Virus (NZB, Xeno, SEQ ID NO: 10), Feline Leukaemia Virus C (FeLV, SEQ ID NO: 19), Env Koala Retrovirus (KoV, SEQ ID NO: 20), Env Porcine Endogeneous Retrovirus-B (Perv B, SEQ ID NO: 22), Human T Leukaemia Virus-1 (HTLV1, SEQ ID NO:27), Human T Leukaemia Virus-2 (HTLV2, SEQ ID NO:28), Human T Leukaemia Virus-4 (HTLV4, SEQ ID NO: 31), and Env Bovine Leukaemia Virus (BLV, SEQ ID NO: 30).

2. The method according to claim 1 , wherein said membrane receptors are selected from the group consisting of CAT1, PiT2, XPR1, SMIT1, PiT1, ASCT1/ASCT2, FLVCR, PAR1, PAR2, and GLUT1.

3. The method according to claim 1 , wherein said target cells are animal stem cells.

4. The method according to claim 3 , wherein said animal stem cells are human stem cells or cancer stem cells.

5. The method according to claim 3 , wherein said target cells are haematopoietic stem cells or B or T cells.

6. The method according to claim 1 , wherein said at least two soluble receptor binding ligands is a set of three to twelve soluble receptor binding ligands isolated from glycoproteins belonging to viruses selected from the group consisting of: Amphotropic MLV (ampho, SEQ ID NO:1), Gibbon Ape Leukemia virus (GALV, SEQ ID NO:2), Feline endogenous virus (RD114, SEQ ID NO:3), vesicular stomatitis virus (VSV, SEQ ID NO:4), Xenotropic Murine Leukaemia Virus (NZB, Xeno, SEQ ID NO: 10), Feline Leukaemia Virus C (FeLV, SEQ ID NO: 19), Env Koala Retrovirus (KoV, SEQ ID NO: 20), Env Porcine Endogeneous Retrovirus-B (Perv B, SEQ ID NO: 22), Human T Leukaemia Virus-1 (HTLV1, SEQ ID NO:27), Human T Leukaemia Virus-2 (HTLV2, SEQ ID NO:28), Human T Leukaemia Virus-4 (HTLV4, SEQ ID NO: 31), and Env Bovine Leukaemia Virus (BLV, SEQ ID NO: 30).

7. The method according to claim 1 , wherein said soluble receptor binding ligand is a set of ten receptor binding ligands wherein the RBD of glycoproteins are selected from glycoproteins belonging to viruses selected from the group consisting of: Amphotropic MLV (ampho), Feline endogenous virus (RD114), Gibbon Ape Leukemia virus (GALV), Xenotropic Murine Leukaemia Virus (NZB, Xeno), Env Koala Retrovirus (KoV), Env Porcine Endogeneous Retrovirus-B (Perv B), Human T Leukaemia Virus-2 (HTLV2), Human T Leukaemia Virus-4 (HTLV4), Env Bovine Leukaemia Virus (BLV), and Env Feline Leukaemia Virus C (FeLV).

8. The method according to claim 1 , wherein said soluble receptor binding ligand is a set of eight receptor binding ligands wherein the RBD of glycoproteins are selected from glycoproteins belonging to viruses selected from the group consisting of: Amphotropic MLV (ampho, SEQ ID NO:1), Gibbon Ape Leukemia virus (GALV, SEQ ID NO:2), Feline endogenous virus (RD114, SEQ ID NO:3), vesicular stomatitis virus (VSV, SEQ ID NO:4), Xenotropic Murine Leukaemia Virus (NZB, Xeno, SEQ ID NO: 10), Feline Leukaemia Virus C (FeLV, SEQ ID NO: 19), Env Koala Retrovirus (KoV, SEQ ID NO: 20), Env Porcine Endogeneous Retrovirus-B (Perv B, SEQ ID NO: 22), Human T Leukaemia Virus-1 (HTLV1, SEQ ID NO:27), Human T Leukaemia Virus-2 (HTLV2, SEQ ID NO:28), Human T Leukaemia Virus-4 (HTLV4, SEQ ID NO: 31), and Env Bovine Leukaemia Virus (BLV, SEQ ID NO: 30).

9. The method according to claim 1 , wherein said RBD of glycoproteins belong to viruses selected from the group consisting of: Amphotropic MLV (ampho), Feline endogenous virus (RD114), Xenotropic Murine Leukaemia Virus (NZB, Xeno), Env Koala Retrovirus (KoV), Env Porcine Endogeneous Retrovirus-B (Perv B), Human T Leukaemia Virus-2 (HTLV2), Human T Leukaemia Virus-4 (HTLV4), and Env Bovine Leukaemia Virus (BLV).

10. The method according to claim 1 , wherein said at least two soluble receptor binding ligands is a set of four soluble receptor binding ligands isolated from glycoproteins belonging to viruses selected from the group consisting of: Amphotropic MLV (ampho, SEQ ID NO:1), Gibbon Ape Leukemia virus (GALV, SEQ ID NO:2), Feline endogenous virus (RD114, SEQ ID NO:3), vesicular stomatitis virus (VSV, SEQ ID NO:4), Xenotropic Murine Leukaemia Virus (NZB, Xeno, SEQ ID NO: 10), Feline Leukaemia Virus C (FeLV, SEQ ID NO: 19), Env Koala Retrovirus (KoV, SEQ ID NO: 20), Env Porcine Endogeneous Retrovirus-B (Perv B, SEQ ID NO: 22), Human T Leukaemia Virus-1 (HTLV1, SEQ ID NO:27), Human T Leukaemia Virus-2 (HTLV2, SEQ ID NO:28), Human T Leukaemia Virus-4 (HTLV4, SEQ ID NO: 31), and Env Bovine Leukaemia Virus (BLV, SEQ ID NO: 30).

11. The method according to claim 1 , for the implementation of a selection process of target cells, having a defined physiological state.

12. The method according to claim 11 , wherein said target cells are stem cells sub-population expressing said at least two membrane receptors.

13. A process of identification and quantification of the expression of membrane receptors to a glycoprotein RBD of target cells comprising the following steps:
a. contacting at least two soluble receptor binding ligands, as defined in claim 1 , optionally marked with a tag, with a target cell to form at least two complexes, each complex being constituted by one said receptor binding ligand and one said membrane receptor of said target cell,
b. identifying each complex formed,
c. quantifying the expression of each membrane receptor of said target cell able to form said complex,
d. optionally, distinguishing receptors expressed to the surface of the membrane of the target cell from the total receptors expressed within the target cell.

14. The process according to claim 13 , wherein said target cells are animal stem cells.

15. The process according to claim 14 , wherein said animal stem cells are human stem cells.

16. The process according to claim 14 , wherein said target cells are haematopoietic stem cells or B cells or T cells.

17. A process of selection of target cells expressing at least one particular membrane receptor to a glycoprotein RBD in a given amount of expression, comprising the following steps:
a. contacting at least two soluble receptor binding ligands, as defined in claim 1 , optionally marked with a tag, with a target cell to form at least two complexes, each complex being constituted

Assignees

• Sitbon Marc
• Battini Jean-Luc
• Taylor Naomi
• Mongellaz Cedric
• Centre Nat Rech Scient
• Univ Montpellier 2

Inventors

• Sitbon Marc
• Battini Jean-Luc
• Taylor Naomi
• Mongellaz Cedric

Classifications

• G01N2400/00

  Assays, e.g. immunoassays or enzyme assays, involving carbohydrates · CPC title

• G01N2333/705

  Assays involving receptors, cell surface antigens or cell surface determinants · CPC title

• G01N33/53 Primary

  Immunoassay; Biospecific binding assay; Materials therefor · CPC title

• G01N33/537 Primary

  with separation of immune complex from unbound antigen or antibody · CPC title