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. A method for producing three or more non-identical antibodies in a single recombinant host cell, the method comprising:
expressing in said single recombinant host cell one or more nucleic acid sequences encoding a common immunoglobulin light chain and at least three different immunoglobulin heavy chains that are capable of pairing with said common immunoglobulin light chain to form functional antigen binding domains to produce three or more non-identical antibodies that comprise said common light chain.
2. The method according to claim 1 , further comprising the step of harvesting the three or more non-identical antibodies from the recombinant host cell or from a culture of said host cell.
3. The method according to claim 1 , wherein at least two of said three or more non-identical antibodies target differing epitopes of the same target antigen.
4. The method according to claim 1 , wherein at least two of said three or more non-identical antibodies have differing affinities for the same target epitope.
5. The method according to 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.
6. The method according to claim 5 , wherein said host cell is obtained from a PER.C6 cell as deposited under ECACC number 96022940.
7. The method according to claim 1 , wherein said three or more non-identical antibodies are independently selected from the group consisting of: IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgD, IgE and IgM.
8. The method according to claim 1 , wherein said at least three different immunoglobulin heavy chains are of IgG isotype.
9. The method according to claim 1 , wherein at least two of said three or more non-identical antibodies bind to different epitopes of one target antigen.
10. The method according to claim 1 , wherein said one or more nucleic acid sequences are stably expressed in said host cell.
11. The method according to claim 1 , wherein said three or more non-identical antibodies are produced by said host cell in vitro.
12. The method according to claim 1 , further comprising:
selecting at least one host cell by assaying said three or more non-identical antibodies produced by said recombinant host cell for their ability to bind a target antigen;
culturing said recombinant host cell; and
isolating said three or more non-identical antibodies.
13. The method according to claim 12 , wherein said host cell is selected using high throughput procedures.
14. The method according to claim 1 , wherein said at least three different immunoglobulin heavy chains have identical constant regions.
15. The method according to claim 1 , wherein at least one of said one or more nucleic acid sequences is integrated in the host cell's genome.
16. The method according to claim 1 , further comprising culturing said host cell for more than 20 population doublings.
17. The method according to claim 1 , wherein said three or more non-identical antibodies comprise monospecific and bispecific antibodies.
18. The method according to claim 1 , wherein at least two of said three or more non-identical antibodies bind to different antigens.
19. The method according to claim 16 , wherein expression of said three or more non-identical antibodies is not substantially diminished over more than 20 population doublings.
20. The method according to claim 1 , wherein said host cell is a mammalian cell.