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Due to their mechanism of action that is different from monoclonal antibodies, double antibodies are widely touted by global pharmaceutical companies.
Hundreds of different antibody formats have been developed so far, mainly divided into two formats, lacking Fc region and having Fc region.
Although each format has specific advantages, antibodies with Fc regions are more prominent in research and clinical settings.
Usually because they are recovered by FcRn, the serum half-life is longer, and the Fc region mediates ADCC and other effector functions, which can enhance anti-tumor activity.
In the blocking application environment, the Fc region may also be modified to reduce or eliminate its ADCC to prevent unwanted effector functions from occurring.
A BsAb carrying an Fc region can generate a symmetrical BsAb by adding an additional binding moiety, such as a single-chain fragment variable (scFv), to the N-terminus or C-terminus of the heavy or light chain.
Alternatively, by introducing mutations in the heavy chain CH3, asymmetric antibodies can be produced.
Asymmetric bispecific antibodies generally have advantages over symmetric bispecific antibodies because they allow monovalent binding to each target.
This is particularly important for T cell redirection methods, because the bivalent binding of CD3 antibodies to T cells may lead to excessive activation and clinically produce immune-related toxicity.
Schematic diagram of the production of symmetric and asymmetric bispecific antibodies (Document 1) The antibody standards suitable for clinical development include: relatively easy production, high stability and good activity.
In order to meet these criteria, many variables must be screened in the research environment.
Therefore, methods to efficiently produce and screen a large number of high-purity antibodies are essential, especially for immune cells involved in BsAb.
Janssen Bipod technology platform Bipod technology platform, one of the binding arms is Fab, and the other is scFv.
To generate these asymmetric BsAbs, one heavy chain contains T350V, L351Y, F405A, and Y407V mutations, and the other heavy chain contains complementary T350V, T366L, K392L, and T394W mutations.
Each chain is expressed by an independent vector, and then a heterodimer molecule is formed by complementary amino acids, and purified based on CH1.
Schematic diagram of Biopod platform (Reference 2) Purification effect analysis is analyzed by SEC and capillary electrophoresis.
Comparison of the purification effect of Protein A and CH1 shows that the purification effect of CH1 is better, and the purity can reach more than 97%.
Functional analysis For a large number of candidate molecules produced by Biopod's high-throughput platform, CD3 and tumor antigen affinity detection, as well as tumor lysis activity and T cell activation analysis, are performed to screen target molecules for further development.
The editor concludes that bispecific antibodies, due to their unique mechanism of action, have shown good prospects in the fields of anti-tumor and autoimmune disease treatment.
However, compared with monoclonal antibodies, the process development of double antibodies is relatively difficult.
In order to better clinical development and subsequent production on the market, double antibodies need to meet the characteristics of easy purification, high stability and moderate activity.
These require a large amount of CMC stage in the early stage.
Research requires a high-throughput method to generate a large number of candidate molecules, and a simplified purification method is required to obtain samples suitable for subsequent research.
Otherwise, the pre-development cycle will be lengthened and the progress of drug development will be delayed.
Roche, Janssen and other international giants have successively tried to develop high-throughput and simplified purification double antibody process platforms to meet the needs of double antibody development.
In addition to Janssen’s Biopod platform introduced in this article, the editor of Biopharmaceuticals also introduced Roche’s FORCE platform before (extended reading: Roche’s FORCE bi-antibody technology: knob-into-hole upgraded version).
Reference 1.
Brinkmann, U.
& Kontermann, RE The making of bispecific antibodies.
MAbs 9, 182–212, https://doi.
org/10.
1080/19420862.
201 6.
1268307 (2017).
2.
Thomas C.
Nesspor et al, High-Throughput Generation of Bipod (Fab × scFv) Bispecific Antibodies Exploits Differential Chain Expression and Affinity Capture, Scientific Reports | (2020) 10:7557 Author l Google Editor l Winston copyright statement welcome personal forwarding and sharing.
Any other media or website that needs to reprint or quote the copyrighted content of this website must be authorized and marked "Reprinted from: Biopharmaceutical Editor" in a prominent position.
Hundreds of different antibody formats have been developed so far, mainly divided into two formats, lacking Fc region and having Fc region.
Although each format has specific advantages, antibodies with Fc regions are more prominent in research and clinical settings.
Usually because they are recovered by FcRn, the serum half-life is longer, and the Fc region mediates ADCC and other effector functions, which can enhance anti-tumor activity.
In the blocking application environment, the Fc region may also be modified to reduce or eliminate its ADCC to prevent unwanted effector functions from occurring.
A BsAb carrying an Fc region can generate a symmetrical BsAb by adding an additional binding moiety, such as a single-chain fragment variable (scFv), to the N-terminus or C-terminus of the heavy or light chain.
Alternatively, by introducing mutations in the heavy chain CH3, asymmetric antibodies can be produced.
Asymmetric bispecific antibodies generally have advantages over symmetric bispecific antibodies because they allow monovalent binding to each target.
This is particularly important for T cell redirection methods, because the bivalent binding of CD3 antibodies to T cells may lead to excessive activation and clinically produce immune-related toxicity.
Schematic diagram of the production of symmetric and asymmetric bispecific antibodies (Document 1) The antibody standards suitable for clinical development include: relatively easy production, high stability and good activity.
In order to meet these criteria, many variables must be screened in the research environment.
Therefore, methods to efficiently produce and screen a large number of high-purity antibodies are essential, especially for immune cells involved in BsAb.
Janssen Bipod technology platform Bipod technology platform, one of the binding arms is Fab, and the other is scFv.
To generate these asymmetric BsAbs, one heavy chain contains T350V, L351Y, F405A, and Y407V mutations, and the other heavy chain contains complementary T350V, T366L, K392L, and T394W mutations.
Each chain is expressed by an independent vector, and then a heterodimer molecule is formed by complementary amino acids, and purified based on CH1.
Schematic diagram of Biopod platform (Reference 2) Purification effect analysis is analyzed by SEC and capillary electrophoresis.
Comparison of the purification effect of Protein A and CH1 shows that the purification effect of CH1 is better, and the purity can reach more than 97%.
Functional analysis For a large number of candidate molecules produced by Biopod's high-throughput platform, CD3 and tumor antigen affinity detection, as well as tumor lysis activity and T cell activation analysis, are performed to screen target molecules for further development.
The editor concludes that bispecific antibodies, due to their unique mechanism of action, have shown good prospects in the fields of anti-tumor and autoimmune disease treatment.
However, compared with monoclonal antibodies, the process development of double antibodies is relatively difficult.
In order to better clinical development and subsequent production on the market, double antibodies need to meet the characteristics of easy purification, high stability and moderate activity.
These require a large amount of CMC stage in the early stage.
Research requires a high-throughput method to generate a large number of candidate molecules, and a simplified purification method is required to obtain samples suitable for subsequent research.
Otherwise, the pre-development cycle will be lengthened and the progress of drug development will be delayed.
Roche, Janssen and other international giants have successively tried to develop high-throughput and simplified purification double antibody process platforms to meet the needs of double antibody development.
In addition to Janssen’s Biopod platform introduced in this article, the editor of Biopharmaceuticals also introduced Roche’s FORCE platform before (extended reading: Roche’s FORCE bi-antibody technology: knob-into-hole upgraded version).
Reference 1.
Brinkmann, U.
& Kontermann, RE The making of bispecific antibodies.
MAbs 9, 182–212, https://doi.
org/10.
1080/19420862.
201 6.
1268307 (2017).
2.
Thomas C.
Nesspor et al, High-Throughput Generation of Bipod (Fab × scFv) Bispecific Antibodies Exploits Differential Chain Expression and Affinity Capture, Scientific Reports | (2020) 10:7557 Author l Google Editor l Winston copyright statement welcome personal forwarding and sharing.
Any other media or website that needs to reprint or quote the copyrighted content of this website must be authorized and marked "Reprinted from: Biopharmaceutical Editor" in a prominent position.
