METHOD FOR SELECTING A SINGLE CELL EXPRESSING A HETEROGENEOUS COMBINATION OF ANTIBODIES
First Claim
1. A method for selecting a single recombinant cell that expresses a heterogeneous combination of monospecific and bispecific antibodies, or antibody fragments thereof, wherein the monospecific and bispecific antibodies, or antibody fragments thereof, are human, humanized, or deimmunized, and wherein the heterogeneous combination has specific affinity for two target epitopes, the method comprising:
- carrying out a process for producing the heterogeneous combination;
the process comprising providing three different variable regions consisting of two heavy chain regions and one light chain variable region in recombinant cells, wherein antigen binding parts of the variable regions originate from a single species and wherein one variable region is able to functionally pair with more than one other variable region, and under conditions allowing for pairing of variable regions and secretion of the paired regions from the recombinant cells resulting in the production of said heterogeneous combination,providing two target epitopes, andselecting a single recombinant cell from the recombinant cells that produces a heterogeneous combination that binds the two target epitopes,wherein the one light chain variable region comprises a rearranged human immunoglobulin light chain variable region consisting essentially of a germline V and a germline J and encoded by a human immunoglobulin light chain V gene segment joined to a human immunoglobulin light chain J gene segment.
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Abstract
Described are 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. Further provided are methods 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.
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Citations
19 Claims
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1. A method for selecting a single recombinant cell that expresses a heterogeneous combination of monospecific and bispecific antibodies, or antibody fragments thereof, wherein the monospecific and bispecific antibodies, or antibody fragments thereof, are human, humanized, or deimmunized, and wherein the heterogeneous combination has specific affinity for two target epitopes, the method comprising:
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carrying out a process for producing the heterogeneous combination;
the process comprising providing three different variable regions consisting of two heavy chain regions and one light chain variable region in recombinant cells, wherein antigen binding parts of the variable regions originate from a single species and wherein one variable region is able to functionally pair with more than one other variable region, and under conditions allowing for pairing of variable regions and secretion of the paired regions from the recombinant cells resulting in the production of said heterogeneous combination,providing two target epitopes, and selecting a single recombinant cell from the recombinant cells that produces a heterogeneous combination that binds the two target epitopes, wherein the one light chain variable region comprises a rearranged human immunoglobulin light chain variable region consisting essentially of a germline V and a germline J and encoded by a human immunoglobulin light chain V gene segment joined to a human immunoglobulin light chain J gene segment. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A method for selecting a single recombinant cell that expresses a heterogeneous combination of monospecific and bispecific antibodies, or antibody fragments thereof, wherein the monospecific and bispecific antibodies, or antibody fragments thereof, are human, humanized, or deimmunized, and wherein the heterogeneous combination has specific affinity for two target epitopes, the method comprising:
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producing a set of antibody combinations from a set of recombinant cells by a process comprising; pairing, in the set of recombinant cells, three different variable regions consisting of two heavy chain variable regions, and one light chain variable region comprising a rearranged human immunoglobulin light chain variable region encoded by a human immunoglobulin light chain V gene segment joined to a human immunoglobulin light chain J gene segment, wherein antigen binding parts of the variable regions originate from a single species and wherein one variable region is able to functionally pair with more than one other variable region, and secreting the paired regions from the recombinant cells to produce an antibody combination, testing the set antibody combinations to identity a single heterogeneous combination that has specific affinity for the two target epitopes, and isolating from the set of recombinant cells a single recombinant cell that produces the single heterogeneous combination.
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Specification