Journal: EBioMedicine

Article Title: Rapid detection of bacterial infection using a novel single-tube, four-colour flow cytometric method: Comparison with PCT and CRP

doi: 10.1016/j.ebiom.2021.103724

Figure Lengend Snippet: A schematic diagram on how to calculate the FCBI-index value of the patient with microbiologically confirmed Streptococcus pneumoniae pneumonia using the novel FCBI-index method. (a) The incubation of the whole blood sample with the receptor-specific antibodies, red blood cell lysis, and flow cytometry run were carried out one after another in the same test tube. (b) During the flow cytometry run, the leukocyte population within gate R1 was first gated out from the side scatter (SSC)/MHCI bivariate histogram, after which gates R2 (red) and R3 (blue) were set around neutrophil and lymphocyte populations, respectively, in the SSC/CD64 bivariate histogram containing leukocytes inside gate R1. The stop count of 1000 monocytes inside the monocyte gate (black/R4) was used, leading to the collection of a total of 5000–20000 leukocytes. (c) For the purpose of data analysis, three additional bivariate histograms, namely SSC/CD35, SSC/CD64, and SSC/CD329 were made, showing mean fluorescence intensities (MFI, which correlate with the number of receptors on the cell surface) of gated neutrophil (red) and lymphocyte (blue) populations. In the SSC/CD35 histogram, B-lymphocytes expressing CD35 were left outside the lymphocyte gate. (d) The actual FCBI-index value was calculated by substituting the obtained receptor-specific MFI data from neutrophil and monocyte populations into the bacterial infection algorithm. A more detailed step-by-step description of the development of the FCBI-index method is presented in the method section and Supplementary Figs. 1–5. 1.54 × gCMF-HBSS: 1.54-fold calcium- and magnesium-free HBSS -buffer supplemented with 1.54 mg/mL gelatin.

Article Snippet: Fluorescence-conjugated receptor-specific monoclonal antibodies (test kit): Phycoerythrin (PE)-conjugated anti-human CD35 (Clone E11) was purchased from Ancell (ANC-184-050), PerCP-Vio700-conjugated anti-human CD64 (Clone 10.1.1) was purchased from Miltenyi Biotec (130-101-422), fluorescein isothiocyanate (FITC)-conjugated anti-human CD329 (Clone 191240) was purchased from R&D systems (FAB1139F) and AF647-conjugated anti-human MHC class I molecule (Clone W6/32) was purchased from R&D systems (FAB7098R). (Supplementary Table 5).

Techniques: Incubation, Red Blood Cell Lysis, Flow Cytometry, Fluorescence, Expressing, Infection

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD64 , REA987 , 50 , 130-116-195 , FITC , Miltenyi Biotec.

Techniques: Imaging

Journal: iScience

Article Title: Antibody attributes, Fc receptor expression, gestation and maternal SARS-CoV-2 infection modulate HSV IgG placental transfer

doi: 10.1016/j.isci.2023.107648

Figure Lengend Snippet:

Article Snippet: FcγRI-activating (human) Reporter Bioassay , Promega , GA1341.

Techniques: Virus, Recombinant, Plasmid Preparation, Molecular Weight, Enzyme-linked Immunosorbent Assay, Bioassay, Binding Assay, Software

Journal: Journal of Nuclear Medicine

Article Title: The Impact of FcγRI Binding on Immuno-PET

doi: 10.2967/jnumed.118.223636

Figure Lengend Snippet: Antibody structure and FcγRI binding. (A) Detailed structure of an antibody with a magnified view of the glycans. (B) Cartoon depicting the influence of deglycosylation on the structure of the Fc region of an antibody and its binding to FcγRI.

Article Snippet: With respect to human and murine FcγRI, a histidine-tagged variant of recombinant human FcγRI (huFcγRI) (500238; NovoPro Labs) was captured as the ligand on a Series S sensor chip NTA (BR-1005-32; GE Healthcare).

Techniques: Binding Assay

Journal: Journal of Nuclear Medicine

Article Title: The Impact of FcγRI Binding on Immuno-PET

doi: 10.2967/jnumed.118.223636

Figure Lengend Snippet: Flow cytometry and ELISA analyses of DFO-bearing immunoconjugates. (A) Flow cytometry with BT474 cells to verify HER2-binding. (B) ELISA of the immunoconjugates (0.5 or 50 μg/mL) with huFcγRI (10 μg/mL) and mFcγRI (10 μg/mL). (C) Flow cytometry of the immunoconjugates with human U937 macrophages and mouse RAW 264.7 macrophages. Data represent mean ± SD; each immunoconjugate was tested in triplicate (distinct samples). MFI = mean fluorescence intensity. Data were analyzed by unpaired, 2-tailed Student t test: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: With respect to human and murine FcγRI, a histidine-tagged variant of recombinant human FcγRI (huFcγRI) (500238; NovoPro Labs) was captured as the ligand on a Series S sensor chip NTA (BR-1005-32; GE Healthcare).

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Binding Assay, Fluorescence

Journal: Journal of Nuclear Medicine

Article Title: The Impact of FcγRI Binding on Immuno-PET

doi: 10.2967/jnumed.118.223636

Figure Lengend Snippet: SPR analysis of interaction between the immunoconjugates and recombinant huFcγRI. (A) Binding affinity and kinetic rate constants for each immunoconjugate are shown in sensorgram plots. (B) Bar graphs illustrating correlation between deglycosylation and binding affinity (KD), half-life (t1/2), on-rate (ka), and off-rate (kd). Statistically significant relationships are indicated with asterisks. * = P < 0.05, ** = P < 0.005, *** = P < 0.0005, **** = P < 0.00005, ***** = P < 0.000005.

Article Snippet: With respect to human and murine FcγRI, a histidine-tagged variant of recombinant human FcγRI (huFcγRI) (500238; NovoPro Labs) was captured as the ligand on a Series S sensor chip NTA (BR-1005-32; GE Healthcare).

Techniques: Recombinant, Binding Assay