Journal: Journal of Virological Methods

Article Title: A high-throughput inhibition assay to study MERS-CoV antibody interactions using image cytometry

doi: 10.1016/j.jviromet.2018.11.009

Figure Lengend Snippet: Exogenously expressed DPP4 in various cell lines. Fluorescence images (A) and fluorescence intensity FlowJo analysis histograms (B) comparing untransfected A549, BHK-21, or Vero E6 cells (left panels) to cells expressing optimal amounts of DPP4, depicted by red staining (right panels).

Article Snippet: Cell nuclei were stained with DAPI, and DPP4 receptors were stained with DPP4 PE-conjugated mouse antibody (Sino Biological, Wayne, PA) or unconjugated DPP4 rabbit polyclonal anitbodies (Abcam, Cambridge, MA) followed by anti-rabbit antibody labeled with Alexa Flour® 488 (AL488) (Abcam).

Techniques: Fluorescence, Expressing, Staining

Journal: Journal of Virological Methods

Article Title: A high-throughput inhibition assay to study MERS-CoV antibody interactions using image cytometry

doi: 10.1016/j.jviromet.2018.11.009

Figure Lengend Snippet: MERS-CoV S1 and S proteins binding to DPP4-expressing cells. Whole well fluorescence images of MERS-CoV S1 (A) and MERS-CoV S (B) proteins binding DPP4-expressing BHK-21 cells. Both S1 monomer and S trimer proteins were stained with anti-MERS-CoV S polyclonal AL488, demonstrating binding to DPP4-expressing cells in green. FlowJo analysis of the image-based fluorescence intensity data for MERS-CoV S1 monomer (C) and S trimer (D) showed peak shifts resulting from binding.

Article Snippet: Cell nuclei were stained with DAPI, and DPP4 receptors were stained with DPP4 PE-conjugated mouse antibody (Sino Biological, Wayne, PA) or unconjugated DPP4 rabbit polyclonal anitbodies (Abcam, Cambridge, MA) followed by anti-rabbit antibody labeled with Alexa Flour® 488 (AL488) (Abcam).

Techniques: Binding Assay, Expressing, Fluorescence, Staining

Journal: Journal of Virological Methods

Article Title: A high-throughput inhibition assay to study MERS-CoV antibody interactions using image cytometry

doi: 10.1016/j.jviromet.2018.11.009

Figure Lengend Snippet: Ability of mAbs to inhibit MERS-CoV S protein binding to DPP4-expressing cells . Inhibition of MERS-CoV S binding to DPP4 by D12 mAb was measured by image cytometry (A) and fluorescence intensity was analyzed by FlowJo histograms (B). As a negative control, a non-relevant isotype mAb was used. Dose response curves of Log mAb concentration versus percent inhibition were generated for D12 (C), G2 (D), and G4 (E) mAbs. (C–E) Each dot represents duplicates. Error bars represent SEM.

Article Snippet: Cell nuclei were stained with DAPI, and DPP4 receptors were stained with DPP4 PE-conjugated mouse antibody (Sino Biological, Wayne, PA) or unconjugated DPP4 rabbit polyclonal anitbodies (Abcam, Cambridge, MA) followed by anti-rabbit antibody labeled with Alexa Flour® 488 (AL488) (Abcam).

Techniques: Protein Binding, Expressing, Inhibition, Binding Assay, Cytometry, Fluorescence, Negative Control, Concentration Assay, Generated

Journal: Journal of Virological Methods

Article Title: A high-throughput inhibition assay to study MERS-CoV antibody interactions using image cytometry

doi: 10.1016/j.jviromet.2018.11.009

Figure Lengend Snippet: Ability of mAbs to neutralize MERS-CoV in cells exogenously-expressing DPP4. Neutralization of MERS-CoV pseudovirus by D12 (red), G2 (blue), and G4 (black) in DPP4-expressing BHK-21 cells. 50% neutralization titers (IC 50 ) were calculated based on sigmoidal curves. Each dot represents duplicates. Error bars represents SEM.

Article Snippet: Cell nuclei were stained with DAPI, and DPP4 receptors were stained with DPP4 PE-conjugated mouse antibody (Sino Biological, Wayne, PA) or unconjugated DPP4 rabbit polyclonal anitbodies (Abcam, Cambridge, MA) followed by anti-rabbit antibody labeled with Alexa Flour® 488 (AL488) (Abcam).

Techniques: Expressing, Neutralization