Journal: International Journal of Nanomedicine

Article Title: An Immunoreceptor-Targeting Strategy with Minimalistic C3b Peptide Fusion Enhances SARS-CoV-2 RBD mRNA Vaccine Immunogenicity

doi: 10.2147/IJN.S463546

Figure Lengend Snippet: Confirmation of targeting proficiency for mC3 and mFc ligands via immunofluorescence assay (IFA). ( A ) Illustration of the ligands-receptors interactions that were demonstrated by IFA. Following mRNA transfection into 293T cells, receptors were incubated with the cells. Then, RBD-mC3 bound CR1 while RBD-mFc bound FcγR. The RBD-mC3 and RBD-mFc were labeled by anti-RBD monoclonal antibodies. The CR1 and FcγR were marked using anti-CR1 polyclonal antibodies and anti-FcγR monoclonal antibodies, respectively. ( B ) Antibodies targeting CR1 (FITC, green) and RBD-mC3 (Cy5, purple) depicted the respective distributions of CR1 and RBD-mC3. ( C ) Colocalization analysis of RBD-mC3 and CR1. ( D ) Antibodies against FcγR (green) and RBD-mFc (purple) showed the distributions of FcγR and RBD-mFc respectively. The blue color (DAPI) represented the cell nuclei. ( E ) Colocalization analysis of RBD-mFc and FcγR. Three regions of interest (ROI) were analyzed. Pearson’s coefficient (R) was calculated in colocalization analysis. R > 0.8 suggests a very strong correlation.

Article Snippet: Then, the cells were blocked with 10% goat serum (Jackson ImmunoResearch) for 1 hr at room temperature, followed by one-hour incubation of receptor proteins, including recombinant human CD35 protein (CR1) (R&D Systems #5748-CD-050, NE Minneapolis, MN) and mouse CD64/FCGR1 protein (Sino Biological #50086-M08H).

Techniques: Immunofluorescence, Transfection, Incubation, Labeling, Bioprocessing

Journal: Scientific Reports

Article Title: Complement Receptor 1 availability on red blood cell surface modulates Plasmodium vivax invasion of human reticulocytes

doi: 10.1038/s41598-019-45228-6

Figure Lengend Snippet: Description of reticulocyte-enriched red blood cell (reRBC) samples for ex vivo invasion assays.

Article Snippet: The used sCR1 is commercially available (Cat No 5748-CD-050, R&D SYSTEMS, USA) and supplied as lyophilized power in sterile condition without a hazard preservative and constituted in sterile PBS (400 μg/ml) as per manufacturer’s instruction.

Techniques: Ex Vivo

Journal: Scientific Reports

Article Title: Complement Receptor 1 availability on red blood cell surface modulates Plasmodium vivax invasion of human reticulocytes

doi: 10.1038/s41598-019-45228-6

Figure Lengend Snippet: P . vivax invasion is inhibited by soluble recombinant CR1 (sCR1) protein. P . falciparum 3D7 invasion of high CR1-expressing neuraminidase (N)-treated cells in the presence of sCR1 was significantly reduced compared with untreated H-CR1 cells (χ 2 = 55.9, p ≤ 0.001), while P . falciparum 3D7 invasion efficiency did not vary in the presence of sCR1 on untreated cells (N = 2). P . vivax invasion of H-CR1 cells in the presence of sCR1 was significantly inhibited relative to H-CR1 without sCR1 (N = 9 isolates). Invasion inhibition was not observed by BSA (N = 5 isolates) or PBS (N = 1 isolate). Invasion efficiency did not significantly vary in the presence of BSA (N = 5 isolates). Bars indicate invasion relative to untreated cells with SD. *p = 0.007.

Article Snippet: The used sCR1 is commercially available (Cat No 5748-CD-050, R&D SYSTEMS, USA) and supplied as lyophilized power in sterile condition without a hazard preservative and constituted in sterile PBS (400 μg/ml) as per manufacturer’s instruction.

Techniques: Recombinant, Expressing, Inhibition

Journal: Scientific Reports

Article Title: Complement Receptor 1 availability on red blood cell surface modulates Plasmodium vivax invasion of human reticulocytes

doi: 10.1038/s41598-019-45228-6

Figure Lengend Snippet: Immuno-precipitation assay to examine PvRBP-soluble CR1 complex formation. An anti-CR1 monoclonal antibody HB8592 was incubated with soluble recombinant CR1 (sCR1) in the presence of either recombinant PfRH4 (right panel) or PvRBP1a and PvRBP1b (middle panel) or PvRBP2a, PvRBP2b and PvRBP2c (left panel) and protein G Sepharose (right, middle and left panels correspond to three different gel blots). Immuno-precipitation eluates were fractionated on SDS-PAGE and visualized using SimplyBlue SafeStain. Molecular weight markers are shown on the left-hand side in kDa for each panel. Soluble CR1 is labeled as sCR1 and the two antibody chains are labeled as Ab. I: Input, U: Unbound and E: Eluate.

Article Snippet: The used sCR1 is commercially available (Cat No 5748-CD-050, R&D SYSTEMS, USA) and supplied as lyophilized power in sterile condition without a hazard preservative and constituted in sterile PBS (400 μg/ml) as per manufacturer’s instruction.

Techniques: Immunoprecipitation, Incubation, Recombinant, SDS Page, Molecular Weight, Labeling