Method for separating target entities from a sample using a composition of mono-specific tetrameric antibody complexes coupled to a surface
An improved method for the preparation of target specific surfaces and uses thereof is described. In particular, the surfaces are bound to mono-specific tetrameric antibody complexes prior to their addition to a sample containing target entities and separating them from non-target entities.
1. A method of separating target entities from a sample comprising target entities and non-target entities, said method comprising:
(a) providing at least one mono-specific tetrameric antibody complex (TAC) coupled to a surface, wherein the TAC comprises two antibodies, each antibody specifically binding to the same target entities and wherein the TAC does not comprise bi-specific TACs;
(b) contacting the sample with the TAC coupled surface under conditions to allow the antibodies of the TAC coupled surface to specifically bind to the target entities to form a target entities-TAC coupled surface complex; and
(c) separating the target entities-TAC coupled surface complex from the sample to separate the target entities from the non-target entities.
2. The method according to claim 1 , wherein the surface is a flask, column of beads; or particles.
3. The method according to claim 2 , wherein the surface is a particle.
4. The method according to claim 3 , wherein the particle is non-magnetic.
5. The method according to claim 3 , wherein the particle is magnetic.
6. The method according to claim 5 , wherein the target entities bound to the TAC of the TAC coupled magnetic particles are separated by placing said sample into a magnetic field to separate the target entities from the non-target entities.
7. The method according to claim 1 , wherein the target entities are selected from the group consisting of cells, bacteria, viruses, cell organelles, proteins and nucleic acids.
8. The method according to claim 7 wherein the target entities are cells.
9. The method according to claim 8 , wherein the cells are selected from the group consisting of erythrocytes, lymphocytes, monocytes, granulocytes, tumor cells, stem cells, hematopoietic progenitor cells, mesenchymal cells, mammary epithelial cells, neural cells, and endothelial progenitor cells.
10. The method according to claim 9 wherein the cells are granulocytes.
11. The method according to claim 10 wherein the monospecific TAC comprises anti-CD66 antibodies.
12. The method according to claim 9 wherein the stem cells are endothelial stem cells or embryonic stem cells.
13. The method according to claim 9 wherein the cells are erythrocytes.
14. The method according to claim 13 wherein the monospecific TAC comprises anti-glycophorin A antibodies.