Antibodies and processes for preparing the same
View Patent ↗Provided herein are various processes for the improved production of antibody producing organisms, antibody producing tissues, antibody producing cells and antibodies. In certain embodiments, provided herein are methods for rapidly producing antibody producing organisms, tissues, cells and antibodies derived from humans, organisms, plants or cells that are genetically altered to over-express certain proteins.
1. A method for generating a B-cell that expresses antibodies specific for a selected antigen comprising:
contacting an antibody expressing B-cell with (i) the selected antigen and (ii) an exogenous fusion peptide, wherein the fusion peptide comprises (a) a peptide transduction domain, and (b) Myc, under conditions wherein the presence of the fusion peptide and the selected antigen promotes proliferation and survival of the B-cell that expresses antibodies specific for the selected antigen.
2. The method of claim 1 , wherein the B-cell is an anergic B cell.
3. The method of claim 2 , wherein B-cell anergy is broken by contacting the anergic B-cell with the fusion peptide.
4. The method of claim 1 , wherein the fusion peptide comprises the sequence
(SEQ ID NO. 5)
MRKKRRQRRRMDFFRVVENQQPPATMPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQ
QSELQPPAPSEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLE
MVTELLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDSLLSSTES
SPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGSPSAGGHSKPPHSP
LVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNRKCTSPRSSDTEENVKR
RTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKATAYILSVQAEEQKLISEEDLL
RKRREQLKHKLEQLRKGELNSKLEGKPIPNPLLGLDSTRTGHHHHHH,
and
variants thereof in which the portion of the fusion peptide corresponding to Myc retains at least 90% amino acid sequence identity, and wherein the amino acid sequence substitutions are conservative, and the variants retain Myc biological activity comprising promoting proliferation and survival of B-cells.
5. The method of claim 1 , wherein contacting the B-cell that expresses antibodies with the fusion peptide occurs ex vivo.
6. The method of claim 1 , wherein contacting the antibody expressing B-cell with the fusion peptide occurs in an animal selected from the group consisting of primate, rodent, livestock, and domestic pet.
7. The method of claim 6 , wherein the animal expresses the selected antigen.
8. The method of claim 7 , wherein the selected antigen is expressed in hematopoietic stem cells, or cells differentiated from the hematopoietic stem cells, wherein the cells were transduced with a nucleic acid encoding the selected antigen ex vivo.
9. The method of claim 6 , wherein the animal is exposed to the selected antigen by immunization.
10. The method of claim 6 , wherein the animal is a mouse.
11. The method of claim 1 , wherein the selected antigen is a self-antigen.
12. The method of claim 1 , wherein the selected antigen is from a virus, bacterium, parasite, or fungus.
13. The method of claim 1 , wherein the peptide transduction domain is a peptide transduction domain of TAT.
14. The method of claim 6 , wherein the animal is selected from the group consisting of a rabbit, a sheep, a hamster, a guinea pig, a mouse, a rat, and a chicken.
15. The method of claim 1 , further comprising recovering the B-cell that expresses antibodies to the selected antigen.
16. The method of claim 1 , further comprising recovering the antibodies specific for the selected antigen.
17. The method of claim 1 , wherein the selected antigen is from a neoplastic cell.
18. The method of claim 1 , wherein the antibody is a human antibody.
19. The method of claim 1 , wherein the antibody is a mouse antibody.
20. The method of claim 1 , wherein the B-cell is a human B-cell.
21. The method of claim 1 , wherein the B-cell is a mouse B-cell.
22. A method for culturing a B-cell that expresses antibodies specific for a selected antigen ex vivo comprising: contacting the B-cell that expresses antibodies with (i) a selected antigen and (ii) an exogenous fusion peptide, wherein the fusion peptide comprises (a) peptide transduction domain, and (b) Myc, under conditions wherein the presence of the fusion peptide and the selected antigen promotes proliferation and survival of the B-cell that expresses antibodies specific for the selected antigen ex vivo.
23. The method of claim 22 , further comprising recovering the antibodies specific for the selected antigen.