sc 390147  (Santa Cruz Biotechnology)

Journal: International immunopharmacology

Article Title: Enhanced cytotoxicity in multiple myeloma via T cells armed with bispecific T cell engager targeting B-cell maturation antigen on cancer cells and CD3 on T cells.

doi: 10.1016/j.intimp.2024.113480

Figure Lengend Snippet: Fig. 3. Production and characterization of αBCMA × αCD3 BITEs using SDS-PAGE and immunoblot analysis, flow cytometry, and immunofluorescence. (A) SDS- PAGE and immunoblot analysis of secreted anti-BCMA BITEs in cell culture supernatant, the statistical asterisks indicated significant differences in the amount of secreted BITEs collected from day 1 to day 7, compared to day 0, (B) Binding ability of αBCMA × αCD3 BITEs using flow cytometry against BCMA+ (NCI-H929) and BCMA- (Raji), and CD3+ cells (Jurkat and human activated T cells; huATCs), and (C) Engagements between BCMA+ target cells (KKU-055-BCMA) and huATCs without (control) and with αBCMA × αCD3 BITEs. Statistical significance from 3 independent experiments was analyzed using Student t-test, with p < 0.05 considered significant (* p < 0.05, ** p < 0.01, and **** p < 0.0001).

Article Snippet: The lysates underwent sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), subsequent transfer to nitrocellulose membrane, and staining with anti-BCMA antibody clone D-6 at 1:500 dilution (sc390147, Santa Cruz Biotechnology, TX, USA) in 5 % skim milk overnight at 4 ◦C with gently shaking.

Techniques: SDS Page, Western Blot, Flow Cytometry, Immunofluorescence, Cell Culture, Binding Assay, Control

Journal: International immunopharmacology

Article Title: Enhanced cytotoxicity in multiple myeloma via T cells armed with bispecific T cell engager targeting B-cell maturation antigen on cancer cells and CD3 on T cells.

doi: 10.1016/j.intimp.2024.113480

Figure Lengend Snippet: Fig. 4. Purification of anti-BCMA bispecific T cell engagers (αBCMA × αCD3 BITEs) and its cytolytic activity against BCMA-overexpressing cells. (A) Immunoblotting of His-tagged αBCMA × αCD3 BITEs after purification, (B) Determination of αBCMA × αCD3 BITE concentration using His-tag ELISA, (C) Representative images of crystal violet staining, and (D) triplicated data of percent specific lysis of αBCMA × αCD3 BITEs in parental KKU-055 (BCMAneg) and BCMA-overexpressing KKU-055 cells. Statistical significance was analyzed using a t-test or two-way ANOVA from 3 independent experiments, with p < 0.05 considered significant (* p < 0.05 and ** p < 0.01).

Article Snippet: The lysates underwent sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), subsequent transfer to nitrocellulose membrane, and staining with anti-BCMA antibody clone D-6 at 1:500 dilution (sc390147, Santa Cruz Biotechnology, TX, USA) in 5 % skim milk overnight at 4 ◦C with gently shaking.

Techniques: Purification, Activity Assay, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Staining, Lysis