Bispecific antibody against human EpCAM and β-Gal - Negative control

Anti-hEpCAM-βGal binds to hEpCAM, but not to CD3

Monoclonal scFv antibody against human EpCAM and β-galactosidase; Negative control

Anti-hEpCAM-βGal is a bispecific antibody that recognizes two sites: human epithelial cell adhesion molecule (hEpCAM) and E. coli β-galactosidase (β-Gal). It is used as a negative control for the Anti-hEpCAM-CD3 bispecific antibody that, like solitomab, engages unstimulated T cells to proliferate and exert cytotoxic activity on EpCAM-positive tumor cells [1].

Human EpCAM (also known as CD326, Trop1) is a transmembrane, 40 kDa glycoprotein highly abundant in almost all human carcinomas, including colon, prostate, ovarian, or breast cancer [2]. In healthy tissue, EpCAM expression is generally low [1,2]. In cancer, EpCAM overexpression is responsible for tumor proliferation, invasion, and migration. Since it is often correlated with a poor prognosis, EpCAM represents an attractive therapeutic target [1].

Key features

• Reacts with human EpCAM
• Unable to interact with human CD3
• cFv of clinical relevant bispecific mAb solitomab
• Hexahistidine (His6) tag
• Provided azide-free
• Each lot is functionally tested

References:

1. Kebenko M, et al., 2018. A multicenter phase 1 study of solitomab (MT110, AMG 110), a bispecific EpCAM/CD3 T-cell engager (BiTE®) antibody construct, in patients with refractory solid tumors. Oncoimmunology. 7(8):e1450710 .
2. Brischwein K, et al., 2006. MT110: a novel bispecific single-chain antibody construct with high efficacy in eradicating established tumors. Mol Immunol. 43(8):1129-43.

Figures

Dose-dependent activation of Jurkat-Lucia™ NFAT cells with Anti-hEpCAM-CD3 in the presence of coated human EpCAM-Fc protein. Human EpCAM-Fc fusion protein (2 µg/ml) was coated on ELISA plates overnight. Jurkat-Lucia™ NFAT cells were added together with Anti hEpCAM-CD3 (red curve), Anti-βGal-hCD3 (grey curve), or Anti-hEpCAM-βGal (blue curve) mAb, alone or in combination (black curve). After 24 hours of incubation, T cell activation was determined by measuring the Lucia luciferase activity using QUANTI-Luc™ detection reagent. Results are presented as fold change over non-induced cells (mean ± SEM).

Specifications

Clonality: Monoclonal bispecific antibody.

Specificity: Targets human EpCAM and E. coli β-galactosidase (β-Gal).

Isotype: none (scFv).

Source: CHO (Chinese hamster ovary) cells.

Purity: >90%. Purified by affinity chromatography.

Tested application: ELISA, flow cytometry

Quality control:

•    The binding of Anti-hEpCAM-βGal to human EpCAM has been confirmed by flow cytometry.
•    The inability of Anti-hEpCAM-βGal to bind to hCD3 has been confirmed using ELISA.
•    The complete sequence of this antibody has been verified.
•    The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells.

Contents

Anti-hEpCAM-βGal purified monoclonal antibody is provided azide-free and lyophilized. It is available in two quantities:

• bimab-ecambg-01: 10 µg
• bimab-epambg-05: 5 x 10 µg

Product is shipped at room temperature.

 Upon receipt, store lyophilized antibody at -20°C.

 Lyophilized product is stable for at least 1 year.