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Journal: mAbs

Article Title: RUBY® – a tetravalent (2+2) bispecific antibody format with excellent functionality and IgG-like stability, pharmacology and developability properties

doi: 10.1080/19420862.2024.2330113

Figure Lengend Snippet: Measurements of melting temperatures (Tm) for different RUBYv9 bsAbs.

Article Snippet: Plates were coated with 0.5 μg/mL antigen, human 5T4 (#ID95677.07, Innovagen) or human CD40 (1493-CD, R&D Systems), in PBS over night at 4°C.

Techniques: Binding Assay

Assessing binding and in vitro functionality of RUBY v9 bsAbs. (a) Dual ELISA showing simultaneous binding of RUBY v9 bsAbs to their respective antigen targets. ELISA plates were coated with one antigen, RUBY bsAbs were added followed by detection using a biotinylated second antigen. (b) The activity of a CD40-EpCAM RUBY v9 bsAb in a CD40 reporter assay. CD40 reporter cells (Promega) were incubated with EpCAM-transfected CHO cells or wild type CHO cells in the presence of the bsAb. CD40 activation was measured according to the manufacturer’s instructions. (c) Demonstration of the agonistic function of the CD137-5T4 RUBY bsAbs v9 bsAb on human CD8 T cells where a dose response dependent IFN-gamma production (absolute values) by human CD8 T cells from one representative individual donor activated with the bispecific constructs in the presence or absence of immobilized 5T4-Fc was observed. Obtained mean (and SD) IFN-gamma levels from one representative individual donor from experiment set 2 is shown.

Journal: mAbs

Article Title: RUBY® – a tetravalent (2+2) bispecific antibody format with excellent functionality and IgG-like stability, pharmacology and developability properties

doi: 10.1080/19420862.2024.2330113

Figure Lengend Snippet: Assessing binding and in vitro functionality of RUBY v9 bsAbs. (a) Dual ELISA showing simultaneous binding of RUBY v9 bsAbs to their respective antigen targets. ELISA plates were coated with one antigen, RUBY bsAbs were added followed by detection using a biotinylated second antigen. (b) The activity of a CD40-EpCAM RUBY v9 bsAb in a CD40 reporter assay. CD40 reporter cells (Promega) were incubated with EpCAM-transfected CHO cells or wild type CHO cells in the presence of the bsAb. CD40 activation was measured according to the manufacturer’s instructions. (c) Demonstration of the agonistic function of the CD137-5T4 RUBY bsAbs v9 bsAb on human CD8 T cells where a dose response dependent IFN-gamma production (absolute values) by human CD8 T cells from one representative individual donor activated with the bispecific constructs in the presence or absence of immobilized 5T4-Fc was observed. Obtained mean (and SD) IFN-gamma levels from one representative individual donor from experiment set 2 is shown.

Article Snippet: Plates were coated with 0.5 μg/mL antigen, human 5T4 (#ID95677.07, Innovagen) or human CD40 (1493-CD, R&D Systems), in PBS over night at 4°C.

Techniques: Binding Assay, In Vitro, Enzyme-linked Immunosorbent Assay, Activity Assay, Reporter Assay, Incubation, Transfection, Activation Assay, Construct