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ACROBiosystems社 Are your CD3 proteins for bispecific antibody research are heterodimers?

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Are your CD3 proteins for bispecific antibody research are heterodimers? 

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2_CD3-targeted therapeutic antibody.jpg 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, CD3E and CD3D can randomly form incorrect heterogeneous complexes, leading to the loss of bioactivity and high batch-to-batch inconsistency.

Here we ACROBiosystems has developed a series of confirmed homogeneous CD3δ/CD3ε and CD3γ/CD3ε proteins with good bioactivity.

※ 1:1 heterodimers verified by nonreductive electrophoresis and MALS
 Show good performance in different applications in therapeutic antibody development
Used for the development of a bispecific antibody in clinical phase successfully

Assay data

※ 1:1 heterodimer verified by Nonreduction electrophoresis and MALS

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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.
 

>>> Protocol

 High bioactivity and low batch differences

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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.

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Designed for different applications of drug development

Application 1: Development and screening of antibody drugs

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Fig.3 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.

>>> Protocol

Application 2: Identification and quality control of antibody drugs

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Fig.4 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) (Cat. No. CDE-M120a) with a linear range of 0.4-6 ng/mL.

>>> Protocol

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Fig.5 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

Application 3: Clinical plasma concentration assay

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Fig.6 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.

>>> Protocol

 

Product List

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