Journal: PLoS ONE

Article Title: Discovery and characterization of potent IL-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy

doi: 10.1371/journal.pone.0211236

Figure Lengend Snippet: (A, C, and E) hybridomas isolated from the alternating antigen approach. (B, D and F) hybridomas isolated from the single antigen approach. (A, B) Binding ELISA results of human and cyno IL-21 cross-reactive hybridoma supernatants. (C, D) Cross-reactive hybridoma supernatants were tested for their ability to inhibit the human IL-21/IL-21R-Fc interaction in a competition ELISA. Numbers on the x-axes are the dilution folds in log scale. The red lines represent the inhibitory activity displayed by IL-21R-Fc and are included for comparison. OD = optical density. (E, F) All cross reactive hybridoma supernatants were also tested for their ability to inhibit IL-21-induced phosphorylation of STAT3 and were assessed by flow cytometry. The red bars represent the neutralizing activity exhibited by soluble IL-21R-Fc blocking the IL-21 interaction and are included for comparison. Light blue bars denote anti-IL-21 clones which neutralize STAT3 phosphorylation.

Article Snippet: Recombinant human and cyno IL-21 and human and cyno IL-21R-Fc were generated at MedImmune (Gaithersburg, MD).

Techniques: Isolation, Binding Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Flow Cytometry, Blocking Assay, Clone Assay

Journal: PLoS ONE

Article Title: Discovery and characterization of potent IL-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy

doi: 10.1371/journal.pone.0211236

Figure Lengend Snippet: The 5 most potent inhibitory hybridoma clones were purified and re-tested in the inhibition assay against (A) human IL-21/IL-21R-Fc and (B) cyno IL-21/IL-21R-Fc interactions. Human PBMCs were stimulated for 15 min with (C) human or (D) cyno IL-21, and phosphorylation of STAT3 was quantified by flow cytometry. Graphs represent inhibition of IL-21–induced STAT3 phosphorylation by the indicated hybridoma clones. Data are representative of 3 donors tested. Percent inhibition was calculated as: 100*(1-(pSTAT3 MFI of stimulated+MEDI7169—pSTAT3 MFI of unstimulated)/(pSTAT3 MFI of stimulated—pSTAT3 MFI from unstimulated)).

Article Snippet: Recombinant human and cyno IL-21 and human and cyno IL-21R-Fc were generated at MedImmune (Gaithersburg, MD).

Techniques: Clone Assay, Purification, Inhibition, Flow Cytometry

Journal: PLoS ONE

Article Title: Discovery and characterization of potent IL-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy

doi: 10.1371/journal.pone.0211236

Figure Lengend Snippet: The 4 most potent inhibitory hybridoma clones were molecularly cloned and expressed as murine/human chimeric IgGs 8B6Chi, 9F11Chi, 9H10Chi, and 19E3Chi, and were characterized for inhibition against (A) human or (B) cyno IL-21/IL-21R-Fc interactions, and (C) epitope bin distribution of the 4 chimeric antibodies.

Article Snippet: Recombinant human and cyno IL-21 and human and cyno IL-21R-Fc were generated at MedImmune (Gaithersburg, MD).

Techniques: Clone Assay, Inhibition

Journal: PLoS ONE

Article Title: Discovery and characterization of potent IL-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy

doi: 10.1371/journal.pone.0211236

Figure Lengend Snippet: MEDI7169 inhibition of human or cyno IL-21/IL-21R-Fc interactions.

Article Snippet: Recombinant human and cyno IL-21 and human and cyno IL-21R-Fc were generated at MedImmune (Gaithersburg, MD).

Techniques: Inhibition