Rapid production of monoclonal antibodies
View Patent ↗The present invention relates to genetically altered hybridomas, myelomas and B cells. The invention also relates to utilizing genetically altered hybridomas, myelomas and B cells in methods of making monoclonal antibodies. The present invention also provides populations of hybridomas and B cells that can be utilized to make a monoclonal antibody of interest.
1. A non-human transgenic animal comprising B cells comprising a transgene, wherein the transgene comprises a nucleic acid encoding Igα and Igβ surface-expressed antibody receptor, wherein the nucleic acid is introduced into the germline of the transgenic animal and is incorporated into the animal's genome, and wherein the transgene is expressed in one or more of immature B cells, mature naïve B cells, mature activated B cells, memory B cells, B lineage lymphocytes and plasma cells, wherein expression of the transgene results in an increase in the level of surface-expressed antibody on the surface of one or more immature B cells, mature naïve B cells, mature activated B cells, memory B cells, B lineage lymphocytes and plasma cells relative to non-transgenic cells.
2. The non-human transgenic animal of claim 1 , wherein the Igα and/or Igβ surface-expressed antibody receptor is mutated.
3. The non-human transgenic animal of claim 1 , wherein at least one surface-expressed antibody receptor is chimeric wherein the chimeric receptor comprises one receptor domain that is derived from a different species of animal that is different than the remaining domains.
4. The non-human transgenic animal of claim 2 , wherein the mutated Igα receptor comprises one or more mutations selected from the group consisting of: Y176F, Y182F, Y193F, and Y204F.
5. The non-human transgenic animal of claim 2 , wherein the mutated Igβ receptor comprises one or more mutations selected from the group consisting of: Y190F and Y206F.
6. A method of generating the non-human transgenic animal of claim 1 comprising:
a) injecting a one or more transgenes comprising one or more nucleic acids encoding Igα and Igβ functionally linked to an expression sequence into an embryo;
b) allowing the embryo to develop into an animal; and
c) crossing the non-human transgenic animal of step (b) with a second animal to produce a third animal.
7. The method of claim 6 , wherein the animal is a mouse.
8. The method of claim 6 , wherein the animal is a rabbit.
9. The method of claim 6 , wherein the animal is a rat.
10. The method of claim 6 , wherein the animal is a guinea pig.
11. The method of claim 6 , wherein the expression of the transgene is inducible.
12. The transgenic animal of claim 1 , wherein the expression of the transgene is controlled by the cre-lox expression system.
13. The transgenic animal of claim 1 , wherein the expression of the transgene is controlled by an inducible tetracycline promoter system.
14. The non-human transgenic animal produced by the method of claim 6 .
15. A B cell obtained from the non-human transgenic animal of claim claim 1 .
16. The non-human transgenic animal of claim 3 , wherein one receptor domain of the chimeric surface-expressed antibody receptor is selected from the group consisting of the human extracellular domain, the human transmembrane domain, and the human intracellular domain.