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With the rise of bispecific drugs, there is a wave of research and development of dual-targeted based on PD-1 / PD-L1 and CTLA-4.

Several researches have showed that anti-CTLA-4 inhibitors increase the expression of PD-L1 in some tumors and the combination of anti-CTLA-4 inhibition and PD-(L)1 inhibition favors effector T cell recruitment in preclinical tumor models.

At present, there are many bispecific drugs in the clinical validation stage, such as MEDI5752, MGD019, KN046 and AK104. And its indications also cover a variety of solid tumors.

Overall, these molecules potential new candidates to simultaneously block PD-1/PD-L1 and CTLA-4/B7 immune checkpoints pathway. hence mimicking the clinical effect of a dual immune checkpoint blockade.

ACROBiosystems has specially launched a series of PD-(L)1 and CTLA-4 homodimer proteins, which are suitable for antibody screening and functional verification, to support the research and development of related targeted drugs. 

Assay Data

 CTLA-4 homodimer structure and purity verified by SELC-MALS

SDS-PAGE and SEC-MALS evaluation for structural homogeneity and purity. The purity of Human CTLA-4, His Tag (Cat. No. CT4-H52H9) is more than 90% using SDS-PAGE under reducing and SEC-MALS. The molecular weight is 45-60 kDa verified by SEC-MALS.

Neutralizing antibody blocking assay

Serial dilutions of Ipilimumab neutralizing antibody (1:2 serial dilutions, from 40 μg/mL to 0.019 μg/mL) were added into Human B7-1, Fc Tag (Cat. No. B71-H5259): Biotinylated Human CTLA-4, Fc,Avitag (Cat. No. CT4-H82F3) binding reactions. The assay was performed according to the above described protocol. Background was subtracted from data points before curve fitting.

Quantitative Analysis of CTLA-4 x OX40 Bispecific Antibody in Human Serum by Intact Assay

Immobilized Human OX40 Protein, His Tag (MALS verified) (Cat. No. OX0-H5224) at 2 μg/mL, add increasing concentrations of CTLA-4 x OX40 bispecific antibody in 50% Human serum and then add Biotinylated Human CTLA-4, His, Avitag (Cat. No. CT4-H82E3) at 0.2 μg/mL. Detection was performed using HRP-conjugated streptavidin with sensitivity of 4 ng/mL (Intact assay, Routinely tested).

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