METHOD FOR SELECTING A SINGLE CELL EXPRESSING A HETEROGENEOUS COMBINATION OF ANTIBODIES
The present invention provides combinations of specific binding proteins, such as immunoglobulins, that are designed to be true combinations, essentially all components of the combination being functional and compatible with each other. The invention further provides a method for producing a composition comprising at least two different proteinaceous molecules comprising paired variable regions, the at least two proteinaceous molecules having different binding specificities, comprising paired variable regions, at least two proteinaceous molecules having different binding specificities, comprising contacting at least three different variable regions under conditions allowing for pairing of variable regions and harvesting essentially all proteinaceous molecules having binding specificities resulting from the pairing.
1 .- 43 . (canceled)
44 . A method for producing nucleic acid sequences encoding variable regions for use in methods for producing a composition comprising at least two different proteinaceous molecules comprising paired variable regions, wherein the at least two different proteinaceous molecules have different binding specificities, the method comprising:
synthesizing nucleic acid sequences encoding variable regions,
expressing the nucleic acid sequences and allowing the expression products to pair, wherein the paired variable regions comprise a first variable region and a second variable region,
selecting nucleic acid sequences encoding variable regions having desired pairing behavior, so as to produce nucleic acid sequences encoding variable regions.
45 . The method according to claim 44 , further comprising altering existing nucleic acid sequences encoding variable regions.
46 . The method according to claim 45 , wherein the alteration increases pairing between variable regions.
47 . The method of according to claim 46 , wherein a nucleic acid sequence encoding a CDR3 domain is altered.
48 . The method according to claim 44 , wherein the variable regions comprise at least one element of variable regions of heavy chains and light chains of antibodies.
49 . The method according to claim 48 , wherein the at least one element comprises at least one complementarity determining region (CDR).
50 . The method according to claim 44 , wherein a nucleic acid sequence is selected encoding a variable region capable of pairing with more than one different variable region.
51 . The method according to claim 44 , wherein a nucleic acid sequence is selected encoding a variable region capable of exclusive pairing with one other variable region.
52 . The method according to claim 50 , further comprising:
expressing the nucleic acid sequence encoding the variable region capable of pairing with more than one different variable region together with nucleic acid sequences encoding the more than one different variable regions.
53 . The method according to claim 52 , wherein the nucleic acid sequence encoding the variable region capable of pairing with more than one different variable region is expressed as a light chain or a fragment thereof; and wherein the nucleic acid sequences encoding the more than one different variable regions are expressed as different heavy chains or fragments thereof.
54 . The method according to claim 53 , wherein the expressed light—and different heavy chains pair so as to produce a heterogeneous combination of antibodies.
55 . The method according to claim 54 , wherein the heterogeneous combination of antibodies has specific affinity for different target epitopes.