BCL2scan: Biochemical ligand binding assays for anti-apoptotic BCL2 family members

Biochemical Ligand Binding Assays for BCL2, BCLXL, BCLW, BCL2A1 and MCL1

DiscoverX has developed a best in class comprehensive panel of quantitative biochemical ligand binding assays for anti-apoptotic BCL2 family proteins that can be used to identify and to optimize therapeutic small molecule inhibitors targeting proteins from this target class. The BCL2 family of proteins determine the commitment of cells to apoptosis and are emerging therapeutic targets for oncology, auto-immune disease, neurodegeneration, and aging-related morbidities. The BCL2 family consists of pro-apoptotic and anti-apoptotic members that interact to regulate the MOMP pathway to apoptosis. The anti-apoptotic family members are the focus of many drug discovery efforts and include BCL2, BCLXL, BCLW, BCL2A1 and MCL1.

The DiscoverX BCL2 family assays enable measurement of potency and selectivity across these anti-apoptotic family members. You can screen, profile and measure Kd values in one flexible format in both high or low-throughput modes.

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BCL2scan Assay List

BGS ▲AliasesBcl2domain Protein NameEntrez Gene Symbol
BCL2Bcl-2; PPP1R50apoptosis regulator Bcl-2 alpha isoform [Homo sapiens]BCL2
BCL2A1GRS; BFL1; ACC-1; ACC-2; HBPA1; BCL2L5BCL2-related protein A1BCL2A1
BCLWBCLW; BCL-W; PPP1R51; BCL2-L-2BCL2-like 2BCL2L2
BCLXLBCLX; BCL2L; BCLXL; BCLXS; Bcl-X; bcl-xL; bcl-xS; PPP1R52; BCL-XL/SBCL2-like 1BCL2L1
MCL1TM; EAT; MCL1L; MCL1S; Mcl-1; BCL2L3; MCL1-ES; bcl2-L-3; mcl1/EATmyeloid cell leukemia 1MCL1
 





References:
  1. Targeting BCL2 with Venetoclax in Relapsed Chronic Lymphocytic Leukemia. Andrew W.Roberts, John Seymour, et al. N ENGL J MED. 2016 Jan 11. 1-12
  2. Clearance of senescent cells by ABT263 rejuvenates aged hematopoietic stem cells in mice. Jianhui Chang, Daohong Zhou et al., Nat.Med. 2016 Jan. 22 (1). 78-83

DiscoverX also offers cell-based services for multiple BCL2 family members including BCL2, BCLXL and MCL1. Learn more

Platform Benefit & Technology Details

  • Menu of fully validated quantitative ligand binding assays for anti-apoptotic BCL2 family targets
  • Measure potency and selectivity in the same assay format enabling inter-protein inhibitor potency rank-ordering and SAR
  • Screen, profile and measure Kds in one flexible format in high and low-throughput modes
  • Broad dynamic range for accurate affinity measurements (tight binding limit <70 pM)

How the Technology Works

Compounds interacting with the BCL2 family protein prevent binding of these proteins to an immobilized known peptide ligand and reduce the amount of target protein captured on the solid support (Panels A & B). Conversely, test molecules that do not bind the BCL2 family protein have no effect on the amount of target protein captured on the solid support (Panel C). Screening “hits” are identified by measuring the amount of BCL2 family protein captured in test versus control samples by using a quantitative, precise and ultra-sensitive qPCR method that detects the associated DNA label (Panel D). In a similar manner, dissociation constants (Kds) for test compound-BCL2 family protein interactions are calculated by measuring the amount of target protein captured on the solid support as a function of the test compound concentration.  Assay conditions are optimized for the measurement of true thermodynamic test compound Kd values.

Available BCL2 Services

  • bcl2KdELECT: Measure test compound Kd values against custom-selected BCL2 family targets
  • bcl2KdMAX: Measure test compound Kd values against all available BCL2 family targets

Standard Turnaround Time: 15 business days for all services. 72 hour TAT available.

Data Analysis & Interpretation

Definitions

Percent of Control (%Ctrl)

The results for single concentration (primary screen) binding interactions for tested compound(s) are reported in your study report and spreadsheets as '%Ctrl' and is calculated in the following manner:

    test compound = client supplied compound
    negative control = DMSO (100% control)
    positive control = control compound (0% control)

Binding Constant (Kd)

The results for an 11-point dose response curve compound/kinase interactions are reported in your study report and spreadsheets as Kd, which are values derived using the Hill equation:
Hill equation used for calculating binding constants
The Hill Slope is set to -1. Curves are fitted using a non-linear least square fit with the Levenberg-Marquardt algorithm.