Journal: Vaccine

Article Title: Fluorescence-barcoded cell lines stably expressing membrane-anchored influenza neuraminidases.

doi: 10.1016/j.vaccine.2025.127157

Figure Lengend Snippet: Fig. 1. Standard NA mAbs brightly and specifically label K530-NA cell lines. Shown are flow cytometry histograms depicting the binding of recombinant IgG versions of standard NA mAbs to K530 cell lines expressing membrane-anchored NAs. Each row corresponds to a monoclonal cell line stably expressing a single type of NA. Each column corresponds to the binding profile for a single mAb. MAbs were incubated with pooled cell lines comprising Option 1 or Option 2 (Table 1), and the resultant data were concatenated into a single figure.

Article Snippet: PBMCs in RPMI-1640 medium plus 10 % FBS were incubated with irrelevant mouse IgG1 (MG1K; Rockland) to block nonspecific binding and then labeled with fluorochrome-conjugated mAbs.

Techniques: Flow Cytometry, Binding Assay, Recombinant, Expressing, Membrane, Stable Transfection, Incubation

Journal: Vaccine

Article Title: Fluorescence-barcoded cell lines stably expressing membrane-anchored influenza neuraminidases.

doi: 10.1016/j.vaccine.2025.127157

Figure Lengend Snippet: Fig. 3. Use of K530-NA cells to determine the binding breadth of newly identified NA mAbs. Shown are flow cytometry histograms depicting the binding of re combinant IgG versions of NA mAbs from donors T1, T2, or T3 to K530 cell lines expressing membrane-anchored NAs, as in Fig. 1.

Article Snippet: PBMCs in RPMI-1640 medium plus 10 % FBS were incubated with irrelevant mouse IgG1 (MG1K; Rockland) to block nonspecific binding and then labeled with fluorochrome-conjugated mAbs.

Techniques: Binding Assay, Flow Cytometry, Expressing, Membrane

Journal: Vaccine

Article Title: Fluorescence-barcoded cell lines stably expressing membrane-anchored influenza neuraminidases.

doi: 10.1016/j.vaccine.2025.127157

Figure Lengend Snippet: Fig. 4. Analysis of serum IgGs elicited by infection of rhesus macaques with H3N2 influenza virus. A) Pre-immune (day 0) or immune (day 27 or 28) blood plasma from three rhesus macaques (6451, T651, T771) infected with A/Aichi/2/1968 (H3N2) influenza virus was incubated with K530-NA cell lines. The degree of IgG labeling of each cell line was determined by flow cytometry, as in Fig. 1. Vertical, black lines denote the threshold for specific labeling of cell-surface NA, determined according to the labeling intensity observed for K530 cells expressing no NA (top row). B) Fold-change in the fluorescence intensity of plasma IgG labeling of selected K530-N2 cell lines, after either infection with H3N2 virus (Fig. 4A) or immunization with recombinant H3 HA (Supplementary Fig. 3B). Fold-change was calculated as the ratio of the geometric mean fluorescence intensity (geoMFI) resulting from labeling with the immune plasma IgG, divided by the geoMFI resulting from labeling with pre-immune plasma IgG. Each symbol represents a single animal. The difference in group means was analyzed by two-tailed t-test with Welch’s correction, as described in Materials and Methods.

Article Snippet: PBMCs in RPMI-1640 medium plus 10 % FBS were incubated with irrelevant mouse IgG1 (MG1K; Rockland) to block nonspecific binding and then labeled with fluorochrome-conjugated mAbs.

Techniques: Infection, Virus, Clinical Proteomics, Incubation, Labeling, Flow Cytometry, Expressing, Fluorescence, Recombinant, Two Tailed Test