Journal: Cell reports

Article Title: Co-immunization of DNA and Protein in the Same Anatomical Sites Induces Superior Protective Immune Responses against SHIV Challenge

doi: 10.1016/j.celrep.2020.107624

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Streptavidin-immobilized CM7 Sensor Chips , GE Healthcare , Cat# 28–953828.

Techniques: Recombinant, Biomarker Assay, Software, Hybridization, Infection, Binding Assay, Spectrophotometry

Journal: Frontiers in Immunology

Article Title: Deregulation of Factor H by Factor H-Related Protein 1 Depends on Sialylation of Host Surfaces

doi: 10.3389/fimmu.2021.615748

Figure Lengend Snippet: α2,3-Linked sialic acid affinity measurements by SPR. The affinities of FH18-20, FHR-1, and their mutant versions for α2,3-linked sialic acid were measured by SPR. All proteins were flowed over a streptavidin sensor chip where approximately 150 RUs of biotin-coupled α2,3-linked sialic acid had been immobilized. (A) 1:1 dilution series of each construct were prepared at concentrations ranging from 0.16 to 40 µM for FH18-20 and its mutants, and 0.04–10 µM for FHR-1 and FHR-1(SV). The corresponding binding curves are shown in blue (40 µM), dark green (20 µM), light green (10 µM), yellow (5 µM), orange (2.5 µM), red (1.25 µM), purple (0.63 µM), dark brown (0.31 µM), light brown (0.16 µM), gray (0.08 µM), and black (0.04 µM). (B) Overlay of binding responses to α2,3-linked sialic acid of all analytes at 10 µM.

Article Snippet: Approximately 2,300 RUs of biotinylated C3b (prepared as previously described) were immobilized on a streptavidin chip (SAP; XanTec bioanalytics), previously conditioned and washed according to the manufacturer’s recommendations.

Techniques: Mutagenesis, Construct, Binding Assay

Journal: Frontiers in Immunology

Article Title: Deregulation of Factor H by Factor H-Related Protein 1 Depends on Sialylation of Host Surfaces

doi: 10.3389/fimmu.2021.615748

Figure Lengend Snippet: Competition of FH and FHR-1. (A) Assessment of competition between FH and FHR-1 on an SPR chip that was immobilized with approximately 430 RU of biotinylated C3b on a streptavidin sensor chip. Either FHR-1, FHR-1(SV), or FH18-20 were injected alone at 3, 7.5, or 50 µM, respectively (approximately 5-fold greater than the determined K D of the respective analytes for biotinylated C3b immobilized onto the SPR chip; see Supplementary Figure 1 ), or in combination with FH at 1 µM (shown in red). (B) Human C3b opsonized RBCs that were either untreated or incubated with neuraminidase (desialylated) were exposed to FH (0.75 µM final concentration) or FH together with increasing FHR-1 or FHR-1(SV) concentrations. FH binding was detected with an anti-FH antibody and a secondary detection molecule with APC fluorescence marker. Data points are mean values (with SD) derived from at least three independent assays.

Article Snippet: Approximately 2,300 RUs of biotinylated C3b (prepared as previously described) were immobilized on a streptavidin chip (SAP; XanTec bioanalytics), previously conditioned and washed according to the manufacturer’s recommendations.

Techniques: Injection, Incubation, Concentration Assay, Binding Assay, Fluorescence, Marker, Derivative Assay