United Kingdom (UK)
Recombinant monoclonal antibody to Melanotransferrin. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma SC57.32.
UniProt Accession Number of Target Protein: P08582
Alternative Name(s) of Target: Melanoma-associated antigen p97; CD228; MTf; MFI2
Immunogen: The original antibody was generated by immunizing Balb/c mice with human Melanotransferrin- His.
Specificity: The Fab fragment is specific for human melanotransferrin (MTf). In particular, it targets the C2 subdomain of the MTf C-lobe, making direct contact with ten MTf residues (Arg 458, Arg 459, Asp 460, Ser 461, His 463, Arg 474, Val 564, Glu 565, Asn 566, Asp 598) as well as two N-acetylglucosamines of the Asn 515 N-glycan. The Fab fragment cross reacts with cynomolgus monkey MTf but not with rat and mouse MTf. Melanotransferrin is a 75 kDa member of the transferrin superfamily, which comprises a single chain, iron-binding glycoproteins responsible for iron homeostasis in cells.
Application Notes: The crystal structure of the Fab fragment in complex with human melanotransferrin (hMFt) was determined. Surface plasmon resonance assay was performed to evaluate the binding specificity and affinity between the Fab fragment and hMFt (Kd= 1.0 nM). The Fab fragment is the murine version of SC-005, which has been used in clinical trials (Hayashi et al., 2021; PMID: 33436675). Immunohistochemistry was performed on HEK293T parental cell pellets or MFI2-expressing HEK293T cell pellets using the murine version of the antibody, which was able to specifically detect MFI2-overexpressing HEK293T cell pellets. The humanized version of the antibody was able to kill HEK-293T cells overexpressing MFI2, showing comparable efficacy to the chimeric antibody. In vivo studies with the humanized version of the antibody conjugated to a pyrrolobenzodiazepine resulted in significant tumor suppression in mice (US10428156).
Antibody first published in:
Hayashi et al. Complex of human Melanotransferrin and SC57.32 Fab fragment reveals novel interdomain arrangement with ferric N-lobe and open C-lobe. Sci Rep. 2021 Jan 12;11(1):566. PMID:33436675
Note on publication:
The paper describes the structure of the Fab fragment in complex with human melanotransferrin.