Journal: PLOS Neglected Tropical Diseases

Article Title: Mayaro virus pathogenesis and immunity in rhesus macaques

doi: 10.1371/journal.pntd.0011742

Figure Lengend Snippet: ( A ) The development of MAYV-binding, IgM and IgG isotype antibody titers were quantified in macaque plasma at 0, 5, 7, and 10 dpi in ELISA. The LOD was a 1:50 plasma dilution with undetectable values graphed as half the LOD. ( B ) The frequency of cells secreting MAYV binding or neutralizing antibodies were quantified in limiting dilution assays in which macaque PBMC from 10 dpi was stimulated with IL-2 and R848 and supernatants were screened in either MAYV ELISAs or MAYV neutralization assays. The LOD frequency was 0.01 cells per 1×10 6 PBMC. ( C ) The binding specificity of MAYV-specific antibodies was characterized in a western blot in which inactivated, purified MAYV was ran on a 4–12% Bis-Tris gel and probed with macaque plasma from 0 or 10 dpi. Blots from only one animal are shown but are representative for all three animals. ( D ) The longitudinal development of MAYV-neutralizing antibodies was quantified in MAYV neutralization assays using heat-inactivated macaque plasma at 0–5, 7, and 10 dpi. 50% plaque reduction neutralization titers (PRNT 50 ) were determined in non-linear regression. The LOD was a 1:20 plasma dilution and undetectable values were graphed as half of the LOD. ( E ) The breadth of antibodies that neutralized other relevant alphaviruses following MAYV infection were characterized in cross-neutralization assays against UNAV, CHIKV, ONNV, RRV, and VEEV using heat-inactivated macaque plasma from 10 dpi. The LOD was a 1:20 plasma dilution and undetectable values were graphed as half of the LOD. ( F ) Antigenic cartography mapping the antigenic distances between viruses is used to visualize the cross-neutralization data. Error bars are SEM in ( A ) and ( E ). A paired t test was used to compare frequency of MBC secreting MAYV binding and neutralizing antibodies in ( B ).

Article Snippet: Plates were washed with ELISA wash buffer before secondary anti-monkey IgG or IgM (H+L) HRP-conjugated detection antibodies (Rockland) were diluted 1:5,000 and added to appropriate plates.

Techniques: Binding Assay, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Neutralization, Western Blot, Purification, Infection

Journal: Marine Drugs

Article Title: In Search of the Role of Three-Finger Starfish Proteins

doi: 10.3390/md22110488

Figure Lengend Snippet: ELISA analysis of the Lystar5 interaction with immobilized membrane fraction of different tissues of A. rubens . Parameters describing the curve fit for coelomocytes: A 1 = 1.97 ± 0.09, EC 50 = 54.5 ± 15.4 nM, nH = 0.71 ± 0.1; for coelomic epithelium: A 1 = 2.85 ± 0.12, EC 50 = 13.4 ± 6.6 nM, nH = 0.75 ± 0.10. Curves were not fitted for the body wall and arm tip. Data present as the background-subtracted data, O.D. ± SEM (n=5).

Article Snippet: Then, plates were blocked by 5% BSA in PBS for 2 h (RT), washed (ELISA wash buffer, Mybiosource, San Diego, CA, USA), incubated with different concentrations of Lystar5 (10 −5 –10 −10 M) for 2 h, washed, incubated with anti-Lystar5 serum (1:100), for 1 h, washed, incubated with donkey anti-mouse HRP-conjugated antibody (Jackson Immunoresearch, 715-035-150, 1:1000) for 1 h, washed, and incubated with TMB-One ELISA Substrate (Mybiosource).

Techniques: Enzyme-linked Immunosorbent Assay, Membrane