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Your Position: Home > CD3 Protein - A hot target for bispecific antibody

CD3 Protein - A hot target for bispecific antibody

CD3 Proteins
Background

A bispecific antibody (BsAb) is a kind of artificial protein that can bind to two different antigens or two epitopes of one antigen simultaneously. Bispecific antibodies can play a synergistic role of the two monoclonal antibodies and have advantages over the simple combination. Because of its unique biological mechanism, the bispecific antibody has become an important branch of therapeutic antibody research and development, and is attracting investments. According to the relevant statistics, there are more than 110 bispecific antibodies in clinical trials and about 180 in pre-clinical development worldwide. Among them, CD3 is the main target[1].

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 recognize the CD3ε in the heterodimer complex to activate the anti-tumor activity of T cells. However, in the process of recombinant expression, CD3E and CD3D can randomly form incorrect heterogeneous complexes, which may lead to the loss of bioactivity and large batch-to-batch inconsistency.


ACROBiosystems has developed a series of homogeneous CD3δ/CD3ε and CD3γ/CD3ε proteins with good bioactivity. They are verified as 1:1 heterodimers by nonreductive electrophoresis and MALS as well. Additionally, they show very good performance in therapeutic antibody screening, identification, characterization as well as clinical pharmacokinetic assays, etc. What’s more, our CD3 products were successfully used for the clinical development of a bispecific antibody, which greatly accelerated the process of therapeutic antibody development.

Product features
MALS verified heterodimer
High binding ability and low batch-to-batch differences
Bioactivity verified by clinical bispecific antibody
Various tags:his,his&Avi,Fc tags, etc.
Data

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.


                       

Fig.1 Human CD3E&CD3D Heterodimer Protein, His Tag&Tag Free (Cat. No. CDD-H52W1) on SDS-PAGE under reducing (R) and non-reducing(NR)condition. The purity of the protein was more than 85% and around 35-43 kDa verified by SEC-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.


                       

Fig.2 Immobilized Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) at 1 μg/mL (100 μL/well) on Streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate, can bind Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3) with a linear range of 0.2-6 ng/mL, and batch differences EC50<0.0001 μg/mL.

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.

Application 1: Development and screening of antibody drugs

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 (Clone: OKT3) with a linear range of 0.2-6 ng/mL.

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 UCHT1 with a linear range of 0.2-6 ng/mL.


Application 2: Identification and quality control of antibody drugs

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 (Clone: OKT3) with a linear range of 0.2-6 ng/mL.

Fig.6 Immobilized Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (Cat. No. CDD-H52W8) at 5 μg/mL (100 μL/well) can bind Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3) with a linear range of 0.4-6 ng/mL.

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 (Clone: OKT3) with an affinity constant of 22.5 nM as determined in a SPR assay (Biacore T200) .

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).

Application 3: Clinical PK assay

Fig.9 Immobilized Human BCMA, His Tag (Cat. No. BCA-H522y) at 2 μg/mL, add increasing concentrations of Bispecific T cell Engager (CD3 X BCMA) in 10% human serum and then add Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) at 0.2 μg/mL. Detection was performed using HRP-conjugated streptavidin with sensitivity of 15 ng/mL.

Product List
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