ACROBiosystems – CD3 – Hot target for bispecific antibody

CD3
BackgroundProduct featuresProduct ListData

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]

Related Page: Introduction of ACROBiosystems
Related Page: Bispecific antibody targets

CD3Function 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.

Product List

Cat. No. Product Description
CDD-H5258 Human CD3E&CD3D Heterodimer Protein, Llama Fc&Llama Fc, low endotoxin
CDD-H52W0 Human CD3 epsilon&CD3 delta Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (MALS verified)Hot
CDD-H52W1 Human CD3 epsilon & CD3 delta Heterodimer Protein, His Tag&Tag Free (MALS verified)Hot
CDD-H52Wa Human CD3 epsilon&CD3 delta Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (MALS verified)
CDD-H82F6 Biotinylated Human CD3 epsilon&CD3 delta Heterodimer Protein, Fc,His,Avitag™&Fc,Flag,Avitag™ (MALS verified)
CDD-H82W0 Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, ultra sensitivity (primary amine labeling) (MALS verified)
CDD-H82W6 Biotinylated Human CD3 epsilon&CD3 delta Heterodimer Protein, His,Avitag™&Tag Free (MALS verified)Hot
CDD-C5259 Cynomolgus CD3E&CD3D Heterodimer Protein, Llama Fc&Llama Fc, low endotoxin
CDD-C52W4 Cynomolgus CD3 epsilon&CD3 delta Heterodimer Protein, His Tag&Flag Tag (MALS verified)
CDD-C52W9 Cynomolgus CD3 epsilon&CD3 delta Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (MALS verified)
CDD-C82W6 Biotinylated Cynomolgus CD3E&CD3D Heterodimer Protein, His,Avitag™&Tag Free (MALS verified)
CDD-M5219 Mouse CD3 epsilon&CD3 delta Heterodimer Protein, His Tag&Flag Tag (MALS verified)
CDD-M52W2 Mouse CD3 epsilon&CD3 delta Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (MALS verified)
CDD-M82W5 Biotinylated Mouse CD3 epsilon&CD3 delta Heterodimer Protein, His,Avitag™&Tag Free
CDD-R5257 Rat CD3 epsilon&CD3 delta Heterodimer Protein, Llama Fc Tag&Llama Fc Tag (MALS verified)
CDD-R52D8 Rat CD3 epsilon&CD3 delta Heterodimer Protein, His Tag&Flag Tag (MALS verified)
CDD-R5259 Rabbit CD3E&CD3D Heterodimer Protein, Llama Fc&Llama Fc, low endotoxin (MALS verified)
CDD-R52D7 Rabbit CD3 epsilon&CD3 delta Heterodimer Protein, His Tag&Flag Tag (MALS verified)
MBS-K003 Human CD3E & CD3D Heterodimer-coupled Magnetic Beads
CDG-H52W5 Human CD3 epsilon&CD3 gamma Heterodimer Protein, His Tag&Tag Free (MALS verified)
CDG-H52W9 Human CD3 epsilon&CD3 gamma Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (MALS verified)Hot
CDG-C5257 Cynomolgus CD3E&CD3G Heterodimer Protein, Fc Tag&Fc Tag (MALS verified)
CDG-C52W6 Cynomolgus CD3E&CD3G Heterodimer Protein, His Tag&Tag Free (MALS verified)
CDG-C8219 Biotinylated Cynomolgus CD3E&CD3G Heterodimer Protein, His,Avitag™&Flag Tag (MALS verified)
CDG-M52D2 Mouse CD3 epsilon&CD3 gamma Heterodimer Protein, His Tag&Flag Tag (MALS verified)
CDG-M52W2 Mouse CD3 epsilon&CD3 gamma Heterodimer Protein,Fc,His Tag&Fc,Flag Tag (MALS verified)
CDG-M58D2 Biotinylated Mouse CD3 epsilon&CD3 gamma Heterodimer Protein, His,Avitag™&Flag Tag (MALS verified)
CDG-R82D4 Biotinylated Rat CD3 epsilon&CD3 gamma Heterodimer Protein, His,Avitag™&Flag Tag (MALS verified)
CDG-R82D3 Biotinylated Rabbit CD3 epsilon&CD3 gamma Heterodimer Protein, His,Avitag™&Flag Tag (MALS verified)
CDG-RB52D5 Rabbit CD3 epsilon&CD3 gamma Heterodimer Protein, His Tag&Flag Tag (MALS verified)
CDE-H5223 Human CD3 epsilon Protein, His Tag
CDE-H5256 Human CD3 epsilon Protein, Fc,His Tag
CDE-H8223 Biotinylated Human CD3 epsilon Protein, His Tag, ultra sensitivity (primary amine labeling)
CDE-H82E1 Biotinylated Human CD3 epsilon Protein, His,Avitag™
CDE-HR2H4 HRP-Human CD3 epsilon Protein, His Tag
CDN-H5253 Human CD3 epsilon Protein, Mouse IgG2a Fc Tag (MALS verified)
CDE-C5226 Cynomolgus CD3 epsilon Protein, His Tag (MALS verified)
CDE-C5254 Cynomolgus CD3 epsilon Protein, Fc,His Tag
CDE-M5256 Mouse CD3 epsilon Protein, Fc Tag
MBS-K026 Human CD3E-coupled Magnetic Beads
CDD-H5224 Human CD3 delta Protein, His Tag
CDD-C52H8 Cynomolgus CD3 delta Protein, His Tag
CDA-H5254 Human CD3 gamma / CD3G Protein, Fc Tag (MALS verified)

More BsAb targets↓

CD3 CD19 CD20 CD38
CD40 CD47 IL-3 R alpha Her2
ErbB3 LAG-3 4-1BB PD-1
PD-L1 CTLA-4 TIM-3 EpCAM
EGF R BCMA PSMA HGF R
ROR1 VEGF R2 FAP IL-1 beta
IL-4 IL-6 R alpha IL-13 IL-17A
CD16 TNF-alpha ANGTP2 OX40
ICOS HGF B7-H3 TGF-beta 1
LRP-5 LRP-6 CEACAM-5 Mucin-1

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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.

purity of CD3

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.

CD3の抗体結合性

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) (Cat. No. CDE-M120a) with a linear range of 0.2-6 ng/mL (QC tested), 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

ELISAへの応用1 ELISAへの応用2

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) (Cat. No. CDE-M120a) 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

ELISAへの応用3 ELISAへの応用4

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) (Cat. No. CDE-M120a) 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-H52W0) at 2 μg/mL (100 μL/well) can bind OKT3 (mouse IgG2a) (Cat. No. CDE-M120a) with a linear range of 10-78 ng/mL.

SPR分析への応用1 SPR分析への応用2

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

臨床PK分析への応用

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.

[1] Siwei N , Zhuozhi W , Maria M C , et al. Biology drives the discovery of bispecific antibodies as innovative therapeutics[J]. Antibody Therapeutics(1):1.

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