Generating multispecific antibody mixtures and methods of uses thereof

Patent metadata

Field	Value
Publication number	US-2019031714-A1
Application number	US-201816042889-A
Country	US
Kind code	A1
Filing date	Jul 23, 2018
Priority date	Jul 21, 2017
Publication date	Jan 31, 2019
Grant date	—

Abstract

The invention relates to the generation of multispecific antibody mixtures include a subset of antibodies isolated from a mixture of two or more monospecific antibodies and one or more bispecific antibodies, wherein all antibodies in the subset have the same common heavy chain. The invention also relates to methods of isolating, purifying, or otherwise producing such a subset of antibodies by using at least one affinity chromatography step. The invention also relates to methods of using such a subset of antibodies in a variety of therapeutic indications.

First claim

What is claimed is:

1 . A subset of antibodies isolated from an antibody mixture that is composed of two or more monospecific antibodies and one or more bispecific antibodies, wherein all antibodies in the subset comprise a common heavy chain.

2 . The subset of claim 1 , wherein the subset consists of antibodies all comprising at least a portion of a kappa light chain.

3 . The subset of claim 1 , wherein the subset consists of antibodies all comprising at least a portion of a lambda light chain.

4 . The subset of claim 1 , wherein the subset consists of antibodies all comprising at least a portion of a kappa light chain and at least a portion of a lambda light chain.

5 . A method of isolating the subset of claim 2 , comprising purifying the subset from an antibody mixture using at least one affinity chromatography step.

6 . The method of claim 5 , wherein the purification step is performed using kappa constant or variable domain specific affinity chromatography media.

7 . A method of isolating the subset of claim 3 , comprising purifying the subset from an antibody mixture using at least one affinity chromatography step.

8 . The method of claim 7 , wherein the purification step is performed using lambda constant specific affinity chromatography media.

9 . A method of isolating the subset of claim 4 , comprising purifying the subset from an antibody mixture using at least a first affinity chromatography step and a second affinity chromatography step.

10 . The method of claim 9 , wherein the first purification step is performed using kappa constant or variable domain specific affinity chromatography media and the second purification step using lambda constant specific affinity chromatography media.

11 . The method of claim 9 , wherein the first purification step is performed using lambda constant specific affinity chromatography media and the second purification step using kappa constant or variable domain specific affinity chromatography media.

12 . A method for isolating a subset of antibodies isolated from an antibody mixture composed of three or more antibodies, wherein at least two antibodies in the mixture are non-identical antibodies, the method comprising:
i) producing an antibody mixture by expressing in a single recombinant host cell one or more nucleic acid sequences encoding a common immunoglobulin heavy chain and at least a first immunoglobulin light chain and a second immunoglobulin light chain, wherein the first and second immunoglobulin light chains are non-identical and each of the first and second immunoglobulin light chains are capable of pairing with the common immunoglobulin heavy chain to form functional antigen binding domains, thereby producing three or more antibodies that comprise the common heavy chain; and ii) isolating a subset of antibodies using at least one affinity chromatography step.

13 . The method of claim 12 , wherein the first immunoglobulin light chain comprises at least a portion of a kappa light chain.

14 . The subset of claim 12 , wherein the second immunoglobulin light chain comprises at least a portion of a lambda light chain.

15 . The subset of claim 12 , wherein the first immunoglobulin light chain comprises at least a portion of a kappa light chain and the second immunoglobulin light chain comprises at least a portion of a lambda light chain.

16 . The method of claim 12 , wherein the isolation step is performed using kappa constant specific affinity chromatography media.

17 . The method of claim 12 , wherein the isolation step is performed using lambda constant specific affinity chromatography media.

18 . The method of claim 15 , wherein the isolation step comprises at least a first affinity chromatography step and a second affinity chromatography step.

19 . The method of claim 18 , wherein the first purification step is performed using kappa constant or variable domain specific affinity chromatography media and the second purification step using lambda constant specific affinity chromatography media.

20 . The method of claim 18 , wherein the first purification step is performed using lambda constant specific affinity chromatography media and the second purification step using kappa constant or variable domain specific affinity chromatography media.

21 . The method of claim 12 , further comprising the step of harvesting the three or more non-identical antibodies from the recombinant host cell or from a culture of the host cell.

22 . The method of claim 12 , wherein at least two of the three or more antibodies target differing epitopes of the same target antigen.

22 . The method of claim 12 , wherein at least two of the three or more antibodies target differing antigens.

23 . The method of claim 12 , wherein the three or more antibodies comprise monospecific and bispecific antibodies.

Assignees

• Novimmune Sa

Inventors

• Fischer Nicolas
• Magistrelli Giovanni
• Masternak Krzysztof

Classifications

• C07K2317/51

  Complete heavy chain or Fd fragment, i.e. VH + CH1 · CPC title

• A61K2039/507

  Comprising a combination of two or more separate antibodies · CPC title

• C07K1/22 Primary

  Affinity chromatography or related techniques based upon selective absorption processes · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61K2039/505

  comprising antibodies · CPC title