EPCAM (Epithelial Cell Adhesion Molecule, CD326) is a 40 kDa glycoprotein involved in cell adhesion, proliferation, and tumorigenesis . Recombinant monoclonal antibodies (mAbs) against EpCAM are produced using advanced genetic engineering to ensure high specificity and reproducibility. These antibodies bind to extracellular epitopes of EpCAM, enabling targeted cancer therapies and diagnostics .
Key methodologies for generating EPCAM recombinant mAbs include:
recEpMab-37: KD = 2.0 × 10⁻⁸ M (CHO/EpCAM) and 3.2 × 10⁻⁸ M (Caco-2 cells) .
Adecatumumab: Binds EpCAM with sub-nanomolar affinity, validated in flow cytometry and immunohistochemistry .
Antibodies targeting the EpCL domain show superior internalization, making them suitable for antibody-drug conjugates (ADCs) .
Immunotoxin scFv2A9-PE: IC₅₀ = 50 pM against EpCAM-positive HHCC cells .
Antibody-Dependent Cellular Cytotoxicity (ADCC): Demonstrated by Adecatumumab in prostate cancer models .
Western Blotting: recEpMab-37 detects EpCAM at 40 kDa in Caco-2 lysates .
Immunohistochemistry: Robust staining of formalin-fixed colorectal carcinoma tissues .
Conformational Sensitivity: Recombinant EpCAM fragments (e.g., EpEX) may lack native folding, reducing antibody binding in ELISA .
Species Cross-Reactivity: Most mAbs are human-specific, limiting preclinical testing in non-primate models .
Bispecific Antibodies: Emerging designs combine EpCAM targeting with immune checkpoint inhibitors to enhance efficacy .