Anti- Gemtuzumab (Anti-Idiotype) Antibody

Detection of Gemtuzumab by Western Blot. Western blot shows Rituximab (negative control), Trastuzumab (negative control), Gemtuzumab, and Brentuximab (negative control) biosimilars. PVDF membrane was probed with 0.5 µg/mL of Anti-Gemtuzumab (Anti-Idiotype) Monoclonal Antibody (Catalog # MAB96171) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody HAF008). A specific band was detected for Gemtuzumab at approximately 240 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Immunoblot Buffer Group 1.

Detection of anti-Siglec-3/CD33 (Gemtuzumab) on U937 Human Cell Line by Flow Cytometry. U937 Human histiocytic lymphoma cell line was stained with anti-Siglec-3/CD33 (Gemtuzumab) (MAB9635), then Anti-Gemtuzumab (Anti-Idiotype) Monoclonal Antibody (Catalog # MAB96171, filled histogram) or Mouse IgG_2AIsotype Control Antibody (MAB003, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (F0101B).

Specificity of Anti-Gemtuzumab (Anti-Idiotype) Antibody in ELISA Capture Assay. Biotinylated biosimilars were serially diluted and incubated with Anti-Gemtuzumab (Anti-Idiotype) Antibody (Catalog # MAB96171, green line) coated at 1 μg/mL on a 96-well polystyrene plate. No cross-reactivity with Rituximab (black line) or biotinylated IgG control (orange line) was observed.

Specificity of Anti-Gemtuzumab (Anti-Idiotype) Antibody in Direct ELISA. A panel of biosimilars was coated at 1 μg/mL in 96-well polystyrene plates. Anti-Gemtuzumab (Anti-Idiotype) Antibody (Catalog # MAB96171, 333 ng/mL) detected Gemtuzumab. No cross-reactivity with Cetuximab, Ibritumomab, Rituximab, Adalimumab, Tositumomab, Brentuximab, Trastuzumab, Solanezumab, Teprotumumab, and Aducanumab was observed.

Anti-Gemtuzumab Bridging Assay. The anti-Idiotype antibody (catalog number MAB96171) directed against Gemtuzumab was coated at 4 μg/mL in a 96-well polystyrene plate to capture the biosimilar Gemtuzumab. Biotinylated Anti-Gemtuzumab used at 0.5 μg/mLwas used as the detection antibody before incubation with Streptavidin-HRP. Other biosimilars did not bridge the anti-idotype antibody, showing specificity of the anti-Gemtuzumab (anti-Idiotype) antibody.