Human Anti-MAGEA3 Recombinant Antibody (clone 21B4) (CAT#: HPAB-0183-LSX)

Figure 1 Binding of recombinant human monoclonal antibodies to epitopes on MAGE A3.

Figure 2 Immunoprecipitation of MAGEA3 by recombinant human monoclonal antibodies.

Figure 3 MAGEA-Family-reactivity of recombinant human monoclonal antibodies.

Recognition of MAGEA, but not of MAGEA4 by recombinant human monoclonal ant ibodies. Human antibodies 21B4, 54B4, 94G11, 9A5, 31H10, 43B1O, 2G4 and 20C10 were used to detect recombinant MAGEA3 (lane A) and recombinant MAGEA4-protein (lane B) in Western Blot analysis.

Figure 4 MAGEA-Family-reactivity of recombinant human monoclonal antibodies.

MAGEA-Family-ELISA. Serial dilutions of human-derived recombinant antibody 21B4 was tested in ELISA for binding to MAGEAl, MAGEA3, MAGEA4 and MAGEA10.

Figure 5 MAGEA-Family-reactivity of recombinant human monoclonal antibodies.

Binding of human-derived recombinant antibodies 21B4, 31H10, 94G1 1 and 54B4 to MAGEA3 and MAGEA6.

Figure 6 EC50 determinat ion of recombinant human monoclonal antibodies to MAGEA3.

Figure 7 Immunohistochemistry analysis of recombinant human monoclonal antibody 21B4 on a selection of human normal and tumor tissues.

A) Normal testis: Selected spermatogonia are stained by 21B4 (arrowheads). No staini g by 2 1 B4 is observed on prostate (B), Lymph node (C), heart tissue (D), and Colon (E). F) Squamous cell carcinoma of the lung: 21B4 specifically stains cancerous infiltrates (circles).

Figure 8 21B4 vs M3H67 in MAG EA-Family-ELISA.

Serial dilutions of human-derived recombinant antibody 21B4 and murine mAb M3H67 were tested in ELISA for binding to MAGEAl, MAGEA3, MAGEA4 and MAGEA10. Antibodies were used in concentrations of 1000 ng/ml 1250 ng/ml, 312.5 ng/ml and 78 ng/ml. 21B4 binds specifically to MAGEA3, M3H67 shows preferential binding to MAGEA4, MAGE1 and MAGEA10. (Y-axis of graph in logarithmical scale.)

Figure 9 21B4 vs M3H67 in Western Blot Analysis.

Protein and MAGEA3 containing cell iysate of SK-MEL-37 cells was separated using Gradient SDS PAGE. Proteins were detected after blotting on nitro cellulose membrane using either 21B4 or mAb M3H67 (mg/ml each antibody). Bound antibody was detected using HRP-labelcd goat anti human IgG (21B4) or goat anti-mouse IgG (M3H67) secondary antibodies. Arrows mark position of MAGEA3.

Figure 10 IHC stainings of cancerous tissue derived from head and neck cancers of three individual patients using either 21B4rc or M3H67.

Case 1 shows an overall similar staining by 21B4rc and M3H67. In Case 2, 21B4rc stains cancerous infiltrates which are not recognized by M3H67 and in case 3 a cancerous infiltrate recognized by M3H67 is not stained by 21B4rc.