Methods and compositions for the production of monoclonal antibodies
View Patent ↗The present invention comprises compositions and methods for making monoclonal antibodies. The present invention further comprises vectors that replicate the immune system components, particularly an antigen-presenting cell (APC) element of the immune synapse. Additionally, the present invention may further comprise synthetic T-cells.
1. A method for the in vitro production of monoclonal antibodies comprising:
a) generating activated CD4+ T cells by incubating T cells with a composition comprising a first, second, and third colloidal metal particle joined together by a scaffolding molecule, wherein a major histocompatibility protein-II (MHC-II) is bound to the first particle, a co-stimulatory B7 protein is bound to the second particle, and a structural protein is bound to the third particle, and wherein an antigen is bound to the MHC-II;
b) generating immunized B cells by incubating B cells with the same antigen as the antigen bound to the MHC-II;
c) generating IgG positive B cells by incubating the activated CD4 + T cells with the immunized B cells; and
d) generating a monoclonal antibody producing cell by immortalizing the IgG positive B cells.
2. The method of claim 1 , wherein the structural protein is selected from the group consisting of; intracellular adhesion molecule (ICAM), LFA-3, and CD72.
3. The method of claim 1 , wherein the colloidal metal particles are the same size.
4. The method of claim 1 , wherein the colloidal metal particles are different sizes.
5. The method of claim 1 , wherein the first colloidal metal particle is 32 nanometers (nm) and the second and third colloidal metal particles are 17 nm.
6. The method of claim 1 , wherein the first, second, and third colloidal metal particle is coated with streptavidin and the scaffolding molecule is a biotinylated protein.
7. The method of claim 6 , wherein the biotinylated protein is human serum albumin.
8. The method of claim 1 , wherein the scaffolding molecule is a di-thiol alkane.
9. The method of claim 1 , wherein the scaffolding molecule is a 2 or 4-arm poly-ethylene glycol (PEG).
10. The method of claim 1 , wherein the colloidal metal is selected from the group consisting of; gold, silver, iron, aluminum, and platinum.
11. The method of claim 1 , wherein the colloidal metal is gold.
12. The method of claim 1 , wherein the composition further comprises pharmaceutically-acceptable component comprising excipients, buffers, carriers, or a combination thereof.
13. The method of claim 1 , wherein the composition further comprises an adjuvant.
14. The method of claim 13 , wherein the adjuvant comprises liposomes, emulsions, microspheres, biodegradable polymers and polystyrene, alum, heat killed M. butyricum and M. tuberculosis, Pertusis toxin, and Tetanus toxin, or LPS and enterotoxin B.
15. The method of claim 1 , wherein the T cells and B cells are of human origin.