In some cases, however, the activity was on the border of the 10 µM cutoff set for separating active from inactive (12 µM for compound ChEMBL73930 and 19 µM for compound ChEMBL258456), and could also point to incoherencies between different assay setups, for example.
DR4-restricted T-cell responses were evaluated with hybridomas mDR17.2 and hDR11.2 [8], [8] specific for 1 using a similar assay setup as for the Aq system (Table 2).
For EPL1, the filter paper assay setup used for CP and Pop1 (Baccelli et al. 2014) was unfortunately not feasible due to excessive foaming of the protein solution (R. Gaderer and V. Seidl-Seiboth, unpublished results).
Application of the merozoite purification method of Boyle et al. [ 18] identified the macrolide antibiotic azithromycin as a candidate inhibitor of P. falciparum asexual blood-stage merozoite invasion of the host erythrocyte (see Fig. 1a for representation of assay setup, Fig. 2 for structure of azithromycin and other drugs used in this study).
In addition to the protocols for assay setup and inoculation onto agar, which were similar to the planktonic assay, the analysis of biofilm activity using the XTT metabolic assay also involved multiple pipetting steps, media exchanges and timed delays that were amenable to automation.
The different cost levels can be thought of as representations of different assay setups, where for example a more complex phenotypic assay is more costly per compound screened compared to a biochemical assay on isolated protein [27, 28].
Like other studies [ 58, 61, 62], we strongly recommend manual clustering for each SNP especially during initial SNP assay setup although it can be time-consuming for large number of SNPs.
Moving forward, PBMC will be rested for 2 h before assay setup.
IC50 values for APO866 were obtained using the same assay setup as described above.
Values that are 13- and 8-fold better that those obtained for the SC50 and LOD of the competitive assay setup with the same antibody.
