United Kingdom (UK)
Recombinant monoclonal antibody to BCMA. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the synthetic clone I09.
UniProt Accession Number of Target Protein: Q02223
Alternative Name(s) of Target: CD269; BCM; TNFRSF17; Tumor necrosis factor receptor superfamily member 17; B-cell maturation protein; 15B2; IO9
Immunogen: The original antibody 15B2 was generated by immunizing six week old female Ablexis transgenic mice with both rHu BCMA-Fc and cynomolgus BCMA-Fc or adherent 293 cells (Ad293) expressing either Hu BCMA or Cyno BCMA. The original mouse antibody 15B2 was mutated to a germline form using primers designed to mutate four non-germline residues in 15B2, which resulted in the generation of this lead optimized antibody.
Specificity: This antibody binds human and cynomolgus monkey BCMA. It acts as a receptor for TNFSF13B/BLyS/BAFF and TNFSF13/APRIL. BCMA expression is highest on terminally differentiated B cells. BCMA is involved in mediating the survival of plasma cells for maintaining long-term humoral immunity. It also activates NF-kappa-B and JNK. The expression of BCMA has been linked to a number of cancers, autoimmune disorders, and infectious diseases.
Application Notes: The binding affinity of this antibody to monomeric soluble human BCMA was determined using ProteOn™ XPR36 instrument. It was reported that the original human IgG1 version of the antibody binds soluble human BCMA with an affinity of kd= 9.82nM. The binding of this antibody to membrane bound human BCMA was evaluated using flow cytometry in multiple myeloma and plasma cell leukemia cell lines that endogenously express BCMA (NCI-H929, MM.1S cell lines and Ad293 expressing human BCMA). This antibody was also used in the generation of a antibody-drug conjugate (ADC) comprising a BCMA monoclonal antibody like I09 conjugated to a cytotoxin to check its anti-tumor effects in the treatment of multiple myeloma (US10988546).
Antibody first published in: PMID:
Flow cytometry using anti-BCMA antibody I09 (Ab03126). Daudi cells were stained with the anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of I09 (Ab03126-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and the cells were analyzed using a FACSCanto flow-cytometer.
Western blot using anti-BCMA antibody I09 (Ab03126). Human spleen (A), tonsil (B), and lymph node (C) tissue lysates (35µg protein in RIPA buffer) were resolved on an SDS-PAGE gel, and blots were probed with the chimeric rabbit version of I09 (Ab03126-23.0) at 2µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used, and protein was detected by chemiluminescence.
Immunofluorescence staining of Daudi cells with anti-BCMA antibody I09 (Ab03126). Immunofluorescence analysis of paraformaldehyde fixed Daudi cells on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of I09 (Ab03126-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing membrane staining. The nuclear stain is DAPI (blue). Panels show, from left-right, top-bottom, Ab03126-23.0, DAPI, merged channels, and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.