cmet  (MedChemExpress)

Journal: mAbs

Article Title: Modular generation of multispecific antibodies using protein trans-splicing

doi: 10.1080/19420862.2025.2573179

Figure Lengend Snippet: Monovalent bispecific IgGs produced by protein splicing recapitulate the functional properties of DuetMab benchmarks in cellular assays. (A) Cartoon representation of the six monovalent bispecific antibodies generated by protein splicing. Each cartoon is positioned directly above the assays in which the molecule was evaluated in subsequent panels. (B) Cellular binding of spliced bispecific antibodies and their DuetMab benchmarks to EGFR + /HER2 + /cMET + NCI-H358 cells and (C) CD3 + HPB-ALL cells. (D) direct, (E) ADC cellular cytotoxicity, and (F) T-cell engager cytotoxicity assays comparing spliced molecules to their DuetMab benchmarks. Nip228 is a non-binding isotype control antibody. Cell binding curves are representative examples from n = 2 experiments. Values for direct, ADC, TCE cytotoxicity represent the mean ± the standard error about the mean of two biological replicates.

Article Snippet: The EGFR+, HER2+, and cMET+ cell line NCI H358 were obtained from the American Type Culture Collection (cat# CRL-5807), and CD3+ cell line HPB-ALL was obtained from DSMZ (cat # ACC 483).

Techniques: Produced, Functional Assay, Generated, Binding Assay, Control

Journal: mAbs

Article Title: Modular generation of multispecific antibodies using protein trans-splicing

doi: 10.1080/19420862.2025.2573179

Figure Lengend Snippet: Production and characterization of monovalent trispecific antibodies by protein splicing. (A) Protein trans-splicing schematic in which an αEGFR-αCD3 Int C -scaffolds and an αHER2 Fab-Int N are reacted to produce a monovalent trispecific antibodies with two distinct geometries. (B) SDS-PAGE analysis of the intein reactants and trispecific antibody products (i, light chain; ii, Fab heavy chain; iii, Int C heavy chain; iv, Fab-Int N ; v, spliced heavy chain). (C) Analytical SEC of the spliced trispecific antibodies and a monovalent bispecific DuetMab. (D) EGFR, HER2, and CD3 antigen binding of the spliced trispecifics compared to monovalent bispecific DuetMab controls measured by biolayer interferometry. (E) Cellular binding of spliced trispecific antibodies to EGFR + /HER2 + NCI-H358 cells and CD3 + HPB-ALL cells. (F) T-cell engager cytotoxicity assays comparing activity of the two geometric orientations of the spliced trispecific antibodies.. Cell binding curves are representative examples from n = 2 experiments. SDS-PAGE, SEC, and BLI measurements are performed post post Ni-NTA cleanup and are representative examples from n = 2 replicates. Values for direct and TCE cytotoxicity represent the mean ± the standard error about the mean of two biological replicates.

Article Snippet: The EGFR+, HER2+, and cMET+ cell line NCI H358 were obtained from the American Type Culture Collection (cat# CRL-5807), and CD3+ cell line HPB-ALL was obtained from DSMZ (cat # ACC 483).

Techniques: SDS Page, Binding Assay, Activity Assay

Journal: mAbs

Article Title: Modular generation of multispecific antibodies using protein trans-splicing

doi: 10.1080/19420862.2025.2573179

Figure Lengend Snippet: Function of bivalent multispecific antibodies produced via dual-splicing. (A) Cartoon representation of the generated multispecific antibodies. Each cartoon is positioned directly above the assays in which the molecule was evaluated in subsequent panels. (B) Cellular binding of spliced multispecific antibodies to EGFR + /HER2 + NCI-H358 cells. (C) Direct and (D) ADC cytotoxicity assays of spliced molecules. (E) Cellular binding to CD3 + HPB-ALL cells. (F) T-cell engager cytotoxicity assay of spliced molecules. Nip228 is a non-binding isotype control antibody. Values for direct cytotoxicity and TCE cytotoxicity represent the mean ± the standard error about the mean of two biological replicates. Values for ADC cytotoxicity represent the mean ± the standard error about the mean of four biological replicates.

Article Snippet: The EGFR+, HER2+, and cMET+ cell line NCI H358 were obtained from the American Type Culture Collection (cat# CRL-5807), and CD3+ cell line HPB-ALL was obtained from DSMZ (cat # ACC 483).

Techniques: Produced, Generated, Binding Assay, Cytotoxicity Assay, Control