Anti-EpCAM mAbs [3-17I](MABL-1499)

MABL-1499

ELISA, FC, IHC

Engineer antibody

Human, Cynomolgus Monkey

3-17I

Recombinant Antibody

This antibody binds human EpCAM. It also cross reacts with Cynomolgus monkey EpCAM.

CD326; Ep-CAM; KS1/4 antigen; KSA; Epithelial cell adhesion molecule; Adenocarcinoma-associated antigen; Cell surface glycoprotein Trop-1; Epithelial cell surface antigen; Epithelial glycoprotein; EGP; Epithelial glycoprotein 314; 17-1A-antigen; EGP314; hEGP314; Major gastrointestinal tumor-associated protein GA733-2; Tumor-associated calcium signal transducer 1; EGP-2; EGP40; KSA; CO17-1A; GA733-2; 3-171

P16422

The original antibody was generated from a human naïve scFv phage display library and later on made into a human IgG1 isotype format.

The antibody specifically interacts with the natural EpCAM+Kato III cell line by flow cytometric analysis. The IgG format of the antibody bound to both human and cynomolgus EpCAM under non-reducing conditions in western blot analysis. In contrast, no binding of the antibody was observed to any reduced EpCAM antigen. Further, ELISA data demonstrated good binding of the scFv and IgG formats of the antibody to both human and cynomolgus EpCAM. The binding affinity of the IgG1 format to human and monkey EpCAM was measured by surface plasmon resonance (Kd= 1 nM and 0.93 nM, respectively). The ability of the IgG format of the antibody to induce ADCC was analyzed using three different breast cancer cell lines: MDA-MB-231, MDA-453, and BT-474. The results clearly demonstrated the antibody-induced ADCC in all three cell lines, MDA-MB-453, MDA-MB-231, and BT-474, in the presence of human PBMCs with respective estimated EC50 values of 0.08 ng/ml, 15 ng/ml, and 0.12 ng/ml. The maximum killings achieved were 75%, 92%, and 61%, respectively. The ability of the IgG version of the antibody to induce complement- dependent cytotoxicity (CDC) was analyzed using two cell lines, KATO III and MT-3. The results clearly demonstrated that the antibody-induced CDC in the cell lines KATO III and MT-3 in the presence of human serum (EC50 = 0.28 ng/ml and 0.38 ng/ml for KATO III and MT-3 cells, respectively) (US8637017B2). Immunohistochemistry studies of the reactivity of the IgG2A format of the antibody were performed on a panel of normal human tissues. The antibody showed a strong reaction to sloughed cells of the esophagus sample, colon, breast, and lung cancers with epithelial origin. The antibody was conjugated to Strepsaporin. PCI of 3–17I-saporin attenuated cellular viability in three different EpCAM-positive cancer cell lines (MCF7, BxPC-3, and WiDr) as measured by MTS assay. Further, PCI of 3–17I-saporin attenuated both the proliferation and colony-forming ability of EpCAM-positive MCF7 cells (Lund et al., 2014; PMID: 24525727).

Lund et al. The novel EpCAM-targeting monoclonal antibody 3-17I linked to saporin is highly cytotoxic after photochemical internalization in breast, pancreas and colon cancer cell lines. MAbs. Jul-Aug 2014;6(4):1038-50. PMID:24525727

Explores the therapeutic potential of this antibody by using 3–17I-saporin conjugate and checking it cytotoxicity after photochemical internalization.

100 μg Purified antibody.

1 mg/ml.

Protein A affinity purified

PBS with 0.02% Proclin 300.

Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.