Blood group antigens of different types for diagnostic and therapeutic applications
The present invention provides compositions and methods for treating or preventing antibody mediated graft rejection and blood typing.
1. A method for determining the presence of anti-blood group antigen reactive antibodies in serum of a subject, said method comprising:
a) providing a collection of blood group antigens coated onto a solid surface, wherein the collection of blood group antigens comprises one or more blood group antigens selected from the group consisting of:
A type 1 GalNAcα1,3(Fucα1,2)Galβ1,3GlcNAcβ1-R,
A type 2 GalNAcα1,3(Fucα1,2)Galβ1,4GlcNAcβ1-R,
A type 3 GalNAcα1,3(Fucα1,2)Galβ1,3GalNAcα1-R,
A type 4 GalNAcα1,3(Fucα1,2)Galβ1,3GalNAcβ1-R,
B type 1 Galα1,3(Fucα1,2)Galβ1,3GlcNAcβ1-R,
B type 2 Galα1,3(Fucα1,2)Galβ1,4GlcNAcβ1-R,
B type 3 Galα1,3(Fucα1,2)Galβ1,3GalNAcα1-R, and
B type 4 Galα1,3(Fucα1,2)Galβ1,3GalNAcβ1-R; wherein the blood group antigens are linked to the solid surface, and R represents the point of attachment to the solid surface;
b) adding said serum from said subject to said blood group antigen coated solid surface;
c) incubating said serum and said blood group antigen coated solid surface for sufficient time for anti-blood group antigen reactive antibodies in said serum to bind to said blood group antigens;
d) removing components in said serum which do not specifically bind to said blood group antigen coated solid surface;
e) incubating said solid surface with ligands capable of specifically binding to said anti-blood group antigen reactive antibodies bound to said blood group antigens; wherein said ligands are labeled with different labels such that one blood group antigen differs from at least one other blood group antigen by being labeled with said differently labeled ligands;
f) removing said labeled ligands that are not bound to said anti-blood group antigen reactive antibodies; and
g) detecting the presence of said differently labeled ligands bound to said anti-blood group antigen reactive antibodies to determine the presence or absence of said reactive antibodies.
2. The method of claim 1 , wherein the labeled ligands are labeled with fluorescent labels.
3. The method of claim 1 , wherein said labeled ligands are antibodies or binding fragments thereof.
4. The method of claim 3 , wherein said labeled ligands are monovalent antibody fragments.
5. The method of claim 4 , wherein said monovalent antibody fragments are Fab or Fab′ fragments.
6. The method of claim 5 , wherein said Fab or Fab′ fragments are selected from the group consisting of an anti-Fc antibody fragment, an anti-kappa light chain antibody fragment, an anti-lambda light chain antibody fragment, and a single chain antibody fragment.
7. The method of claim 1 , wherein the collection of blood group antigens comprises at least one blood group antigen selected from the group consisting of:
A type 1 GalNAcα1,3(Fucα1,2)Galβ1,3GlcNAcβ1-R,
A type 2 GalNAcα1,3(Fucα1,2)Galβ1,4GlcNAcβ1-R,
A type 3 GalNAcα1,3(Fucα1,2)Galβ1,3GalNAcα1-R, and
A type 4 GalNAcα1,3(Fucα1,2)Galβ1,3GalNAcβ1-R.
8. The method of claim 1 , wherein the collection of blood group antigens comprises at least one blood group antigen selected from the group consisting of:
B type 1 Galα1,3(Fucα1,2)Galβ1,3GlcNAcβ1-R,
B type 2 Galα1,3(Fucα1,2)Galβ1,4GlcNAcβ1-R,
B type 3 Galα1,3(Fucα1,2)Galβ1,3GalNAcα1-R, and
B type 4 Galα1,3(Fucα1,2)Galβ1,3GalNAcβ1-R.
9. The method according to claim 1 , wherein the collection of blood group antigens comprises at least two blood group antigens selected from the group consisting of:
A type 1 GalNAcα1,3(Fucα1,2)Galβ1,3GlcNAcβ1-R,
A type 2 GalNAcα1,3(Fucα1,2)Galβ1,4GlcNAcβ1-R,
A type 3 GalNAcα1,3(Fucα1,2)Galβ1,3GalNAcα1-R, and
A type 4 GalNAcα1,3(Fucα1,2)Galβ1,3GalNAcβ1-R.
10. The method according to claim 1 , wherein the collection of blood group antigens comprises at least three blood group antigens selected from the group consisting of:
A type 1 GalNAcα1,3(Fucα1,2)Galβ1,3GlcNAcβ1-R,
A type 2 GalNAcα1,3(Fucα1,2)Galβ1,4GlcNAcβ1-R,
A type 3 GalNAcα1,3(Fucα1,2)Galβ1,3GalNAcα1-R, and
A type 4 GalNAcα1,3(Fucα1,2)Galβ1,3GalNAcβ1-R.
11. The method according to claim 1 , wherein the collection of blood group antigens comprises at least four blood group antigens selected from the group consisting of:
A type 1 GalNAcα1,3(Fucα1,2)Galβ1,3GlcNAcβ1-R,
A type 2 GalNAcα1,3(Fucα1,2)Galβ1,4GlcNAcβ1-R,
A type 3 GalNAcα1,3(Fucα1,2)Galβ1,3GalNAcα1-R, and
A type 4 GalNAcα1,3(Fucα1,2)Galβ1,3GalNAcβ1-R.
12. The method according to claim 1 , wherein the collection of blood group antigens comprises at least two blood group antigens selected from the group consisting of:
B type 1 Galα1,3(Fucα1,2)Galβ1,3GlcNAcβ1-R,
B type 2 Galα1,3(Fucα1,2)Galβ1,4GlcNAcβ1-R,
B type 3 Galα1,3(Fucα1,2)Galβ1,3GalNAcα1-R, and
B type 4 Galα1,3(Fucα1,2)Galβ1,3GalNAcβ1-R.
13. The method according to claim 1 , wherein the collection of blood group antigens comprises at least three blood group antigens selected from the group consisting of:
B type 1 Galα1,3(Fucα1,2)Galβ1,3GlcNAcβ1-R,
B type 2 Galα1,3(Fucα1,2)Galβ1,4GlcNAcβ1-R,
B type 3 Galα1,3(Fucα1,2)Galβ1,3GalNAcα1-R, and
B type 4 Galα1,3(Fucα1,2)Galβ1,3GalNAcβ1-R.
14. The method according to claim 1 , wherein the collection of blood group antigens comprises at least four blood group antigens selected from the group consisting of:
B type 1 Galα1,3(Fucα1,2)Galβ1,3GlcNAcβ1-R,
B type 2 Galα1,3(Fucα1,2)Galβ1,4GlcNAcβ1-R,
B type 3 Galα1,3(Fucα1,2)Galβ1,3GalNAcα1-R, and
B type 4 Galα1,3(Fucα1,2)Galβ1,3GalNAcβ1-R.
15. The method according to claim 1 , where in the solid surface is a microtiter plate, resin, nitrocellulose, or nylon.