CD3 Proteins-Hot target for bispecific antibody

T cell bispecific antibodies (TCBs) are molecules that are engineered to include binding sites to the T cell receptor (TCR) and to tumor-associated or tumor-specific antigens within a single entity. One of the most widely studied TCBs in cancer immunotherapy is CD3, a molecule that has the function of stabilizing TCR structure and transmitting activation signals.

Antibodies against CD3 molecules can stimulate or block T cell activation signal transduction. In addition, these antibodies can eliminate effector T cells or induce a regulatory phenotype in T cells thus, providing a new method for the treatment of organ transplant rejection and autoimmune diseases. These CD3 molecules bind to TCR non-covalently to form a TCR/CD3 receptor complex on the surface of the T cell.

There are 4 subtypes of CD3, including CD3δ, CD3ε, CD3γ, and CD3ζ. CD3δ/CD3ε and CD3γ/CD3ε can form the TCR-CD3 complexes through the α/β chain of TCR as heterodimers. CD3-targeted therapeutic antibody usually recognizes the CD3ε in the heterodimer complex to activate the anti-tumor activity of T cells. However, in the process of recombinant expression, CD3ε and CD3δ can randomly form incorrect heterogeneous complexes, which may lead to the loss of bioactivity and high batch-to-batch inconsistency.

To accelerate the research and development of bispecific antibodies, ACROBiosystems has developed a series of homogeneous CD3δ/CD3ε and CD3γ/CD3ε proteins with high bioactivity. These proteins are verified as a 1:1 heterodimer by nonreductive electrophoresis and MALS respectively. In addition, customer data suggests that bispecific antibody products from ACROBiosystems can be used in applications including, but not limited to clinical pharmacokinetics, therapeutic antibody screening, identification, and characterization.

Successively, our superior CD3 product quality helped accelerate the clinical development of a bispecific antibody into a therapeutic. Click here to download CD3 poster.

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CD3-targeted bispecific and trispecific antibodies represent a fast-growing field in therapeutic antibody development. However, selecting the appropriate detection reagents for custom bioanalytical assays such as ELISAs is challenging. With an abundant selection of CD3 proteins including labels and tags, companies may struggle in developing a highly sensitive and specific assay for their drug therapy.

Here at ACROBiosystems, we can provide a large product catalog of highly bioactive CD3δ/CD3ε, CD3γ/CD3ε products as well as our expertise and our ISO17025-accedited analytical service platform. Using our analytical service platform, we offer customized services from paired protein screening to ELISA-based assay release method development, validation, and kit development to help accelerate your R&D process!

Case Studies

Immobilized Human CD3E&CD3D Heterodimer Protein, Fc Tag&Fc Tag (Cat. No. CDD-H5255) at 1 μg/mL (100 μL/well) was spiked with different concentrations of bispecific BCMA×CD3 T cell-engaging Antibody. The identified linear range was found to be 0.8-6 ng/mL.

Protocol

Immobilized Human BCMA, His Tag (Cat. No. BCA-H522y) was bound to a 96-well plate. Increasing concentrations of bispecific T cell Engager (CD3 X BCMA) spiked in 10% human serum and Biotinylated Human CD3E&3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) at 0.2 μg/mL was added to each well. Detection was performed using HRP-conjugated streptavidin with sensitivity of 15 ng/mL.

Protocol

1:1 heterodimer verified by nonreductive electrophoresis and MALS

ACROBiosystems has developed a series of homogeneous CD3 proteins with good bioactivity, which are verified as 1:1 heterodimer by nonreductive electrophoresis and MALS.

High binding ability and low batch-to-batch differences

These verified authentic 1:1 heterodimer proteins show high bioactivity in different applications. The products are homogenous with strict quality control to guarantee the minimal batch-to-batch differences.

Designed for different applications of drug development

These products are suitable for various applications at different stages of drug development, such as antibody immunotiter detection, biopanning, antibody screening, affinity characterization and clinical pharmacokinetic assays.

Fig.3 Immobilized Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) at 1 μg/mL (100 μL/well) on streptavidin precoated (0.5 μg/well) plate, can bind Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a  (Cat. No. CDE-M120a) with a linear range of 0.2-6 ng/mL.

Protocol

Fig.4 Immobilized Human CD3E&CD3D Heterodimer Protein, His Tag&Tag Free (Cat. No. CDD-H52W1) at 1 μg/mL (100 μL/well) can bind Anti-CD3 antibody with a linear range of 0.2-6 ng/mL.

Protocol

Fig.5 Immobilized Human CD3E&CD3D Heterodimer Protein, His Tag&Tag Free (Cat. No. CDD-H52W1) at 1 μg/mL (100 μL/well) can bind Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a  (Cat. No. CDE-M120a) with a linear range of 0.2-6 ng/mL.

Protocol

Fig.6 Immobilized Human CD3E&CD3D Heterodimer Protein, Fc Tag&Fc Tag (Cat. No. CDD-H5255) at 1 μg/mL (100 μL/well) can bind Bispecific BCMA×CD3 T cell-engaging Antibody with a linear range of 0.8-6 ng/mL.

Protocol

Fig.7 Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) captured on Biotin CAP - Series S sensor Chip can bind Anti-Human CD3 Antibody, Mouse IgG2a  with an affinity constant of 22.5 nM as determined in a SPR assay (Biacore T200) .

Protocol

Fig.8 Bispecific T-cell Engager (CD3 X BCMA) immobilized on CM5 Chip can bind Human CD3E&CD3D Heterodimer Protein, His Tag&Tag Free (Cat. No. CDD-H52W1) with an affinity constant of 31.8 nM as determined in a SPR assay (Biacore T200).

Protocol