Phosphohistidine and phosphotyrosine analogues

Patent metadata

Field	Value
Publication number	US-10053476-B2
Application number	US-201414914994-A
Country	US
Kind code	B2
Filing date	Sep 2, 2014
Priority date	Sep 4, 2013
Publication date	Aug 21, 2018
Grant date	Aug 21, 2018

Abstract

The invention relates to phosphohistidine analogs. The invention also relates to antibodies that specifically bind to the analogs and methods of generating said antibodies. In one embodiment of the invention there is provided a phosphohistidine analog of Formula V: (V) wherein W is selected from H, CO 2 H or CONH 2 ; and X is selected from CH or N.

First claim

The invention claimed is:

1. A phosphohistidine analogue of Formula V or Formula III:

wherein
W is selected from H, CO 2 H or CONH 2 ;
X 1 is CH;
X 2 is selected from NH, O or S.

2. The phosphohistidine analogue according to claim 1 , wherein said analogue is selected from the group consisting of:

3. The phosphohistidine analogue according to claim 1 , wherein the analogue is:

4. The phosphohistidine analogue of claim 1 wherein W is selected from H or CO 2 H.

5. The phosphohistidine analogue according to claim 3 , wherein W is selected from H or CO 2 H.

6. A polypeptide that is a sequence of amino acids that are joined together through amide bonds, which sequence includes the analogue of claim 1 .

7. A method of producing an anti-phosphohistidine antibody, comprising:
(a) immunizing a non-human animal with an analogue according to claim 1 , wherein said analogue is coupled to a carrier protein;
(b) collecting serum from the immunized animal; and
(c) isolating from the serum a population of polyclonal antibodies that specifically bind to the analogue.

8. The method according to claim 7 , wherein said carrier protein is keyhole limpet hemocyanin (KLH), or bovine serum albumin (BSA).

9. A method of producing an anti-phosphohistidine antibody, comprising:
(a) immunizing a non-human animal with an analogue according to claim 1 , wherein said analogue is coupled to a carrier protein;
(b) generating hybridomas by fusing B-cells isolated from the immunized animal with myeloma cells;
(c) identifying hybridomas that produce monoclonal antibodies that specifically bind to the analogue; and
(d) propagating said identified hybridomas to produce monoclonal antibodies that bind to the analogue.

10. The method according to claim 9 , wherein said carrier protein is keyhole limpet hemocyanin (KLH), or bovine serum albumin (BSA).

11. A polypeptide that is a sequence of amino acids that are joined together through amide bonds, which sequence includes the analogue of claim 2 .

12. A polypeptide that is a sequence of amino acids that are joined together through amide bonds, which sequence includes the analogue of claim 3 .

Assignees

• Univ Sheffield

Inventors

• Jackson Richard
• Muimo Richmond
• Lilley Matthew

Classifications

• G01N33/5308

  for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites · CPC title

• C07F9/587

  Chemistry & Metallurgy · mapped topic

• C07F9/65036 Primary

  Chemistry & Metallurgy · mapped topic

• C07K14/001

  by chemical synthesis · CPC title

• C07F9/6539

  Five-membered rings · CPC title