We speculate the on/off PD173074 BI253 Pazopanib charge from the AVL-292 derivative versus AVL-292 may well
vary in cells stimulated with mega CD40L; having said that, even more
investigation is needed.
From a phenotypic point of view, the EPIC B cell activation
assay is made to determine inhibitors that target acknowledged pro-
teins as well as other novel mechanisms of action. Nevertheless,
deconvoluting the hits post HTS poses a challenge. Probably an
attribute of the EPIC assay that distinguishes it from the FLIPR
cell-based assay may be the kinetic profile of the offered compound. For
instance, it could be feasible to further group HTS hits based mostly on
their kinetic trace profile. From a therapeutic standpoint,
examining kinetic profiles of B cell inhibitor medication with desir-
in a position and undesirable properties may perhaps supply a ��profile signa-
ture�� that may be applied to group inhibitors of B cell activation
Nonetheless, appropriate follow-up assays should be
in place to validate this hypothesis. Profiling medication for any ��sig-
nature�� or ��fingerprint�� has been addressed while in the high-con-
tent-imaging arena. Such as, Anne Carpenter��s lab on the
Broad Institute used image-based profiling of a myriad of cel-
lular morphological responses in response to small-molecular
treatment method making use of CellProfiler application. Each the phenotypic
image-based plus the EPIC-based PD173074 BI253 Pazopanib approaches could deliver valuable insights for predicting a compound��s mechanism of
action in the target-agnostic paradigm.
In summary, the two the EPIC LFA-1/ICAM-1 adhesion
assay as well as FLIPR Ca
assay can identify inhibitors of B
The FLIPR-based assay is a lot more amenable to
ultra-HTS in contrast to your EPIC assay; having said that, given that
the readout in the EPIC assay is additional downstream than
the FLIPR-based Ca
release, we anticipate the EPIC
assay will recognize far more inhibitors with differing mecha-
nisms of action. Additionally, the EPIC assay appears much more
sensitive to unique approaches of activating B cells (i.e.,
mega-CD40L/anti-IgM) when in contrast for the FLIPR
assay. Each phenotypic assays are complementary to every
other, along with the decision of platform will largely rely upon the
biological question to become addressed.
We acknowledge Nidhi Arora for PD173074 BI253 Pazopanib thoughtful discussions.Declaration of Conflicting Interests
The authors declared no likely conflicts of curiosity with respect
on the analysis, authorship, and/or publication of this post.
The authors obtained no financial support to the analysis, author-
ship, and/or publication of this post.
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