Recombinant production of mixtures of antibodies
The invention provides methods for producing mixtures of antibodies from a single host cell clone, wherein, a nucleic acid sequence encoding a light chain and nucleic acid sequences encoding different heavy chains are expressed in a recombinant host cell. The recombinantly produced antibodies in the mixtures according to the invention suitably comprise identical light chains paired to different heavy chains capable of pairing to the light chain, thereby forming functional antigen-binding domains. Mixtures of the recombinantly produced antibodies are also provided by the invention. Such mixtures can be used in a variety of fields.
1. An isolated host cell capable of producing three or more non-identical antibodies, said host cell comprising: at least one exogenously introduced nucleic acid sequence encoding an immunoglobulin light chain and exogenously introduced nucleic acid sequences encoding at least three different immunoglobulin heavy chains, wherein said at least three different immunoglobulin heavy chains are capable of pairing with said immunoglobulin light chain to form the three or more non-identical antibodies that bind antigen.
2. The host cell of claim 1 , wherein said at least one exogenously introduced nucleic acid sequence encoding the immunoglobulin light chain is integrated into the host cell's genome.
3. The host cell of claim 1 , wherein said host cell is identified using high throughput procedures.
4. The host cell of claim 1 , wherein said at least one exogenously introduced nucleic acid sequence is present on a low copy number vector and said host cell produces 1 to 20 picograms per cell per day of two or more non-identical antibodies.
5. The host cell of claim 1 , wherein one immunoglobulin light chain is introduced into the host cell.
6. The host cell of claim 1 , wherein said host cell comprises a human embryonic retina cell, and wherein said host cell has been immortalized or transformed by adenoviral E1 nucleic acid sequences.
7. The host cell of claim 1 , wherein said host cell is obtained from a cell as deposited under ECACC number 96022940.
8. The host cell of claim 1 , wherein the three or more non-identical antibodies comprise two or more non-identical antibodies that are of different isotypes.
9. The host cell of claim 8 , wherein said different isotypes comprise at least an IgG and an IgA.
10. The host cell of claim 8 , wherein said different isotypes comprise at least an IgG1 and an IgG3 antibody.
11. The host cell of claim 1 , wherein said three or more non-identical antibodies are of an isotype independently selected from the group consisting of: IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgD, IgE and IgM.
12. The host cell of claim 1 , wherein said three or more non-identical antibodies comprise at least one bispecific antibody.
13. The host cell of claim 1 , wherein said host cell is present in a non-human animal.
14. The host cell of claim 13 , wherein said exogenously introduced nucleic acid sequences encoding the immunoglobulin light and at least three different immunoglobulin heavy chains comprise a tissue-specific promoter.
15. The host cell of claim 1 , wherein said exogenously introduced nucleic acid sequences encoding at least three different immunoglobulin heavy chains are integrated into the host cell's genome.
16. The host cell of claim 1 , wherein at least two of said three or more non-identical antibodies bind to different epitopes of at least one target antigen.
17. The host cell of claim 1 , wherein said at least three different immunoglobulin heavy chains have identical constant regions.
18. The host cell of claim 1 , wherein at least two of said three or more non-identical antibodies bind to different antigens.
19. A culture of host cells producing three or more non-identical antibodies from a single cell within the culture, said culture comprising an isolated host cell according to claim 1 and a culture medium.
20. The culture according to claim 1 , wherein said culture produces said three or more non-identical antibodies from a single cell within the culture for more than 20 population doublings.