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Assay Detail

CHEMBL3885765

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Binding
A2A protein embedded into nanodisc was immobilized via 1D4 antibody on the CM5 sensors. Small molecule ligands were analyzed in multiple cycle kinetic mode. No regeneration was applied.
9
Total Activities
3
Compounds Tested
3
Activity Types
20
Assay Parameters

Assay Information

Assay Type Binding
Organism Homo sapiens
Confidence 9 — Direct single protein target
Curated By Autocuration

Target

Adenosine receptor A2a (CHEMBL251)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

K4DD drug target binding kinetics data
Doris A.Schuetz, Wilhelmus Egbertus Arnout de Witte, Yin Cheong Wong, Bernhard Knasmueller, Lars Richter, Daria B. Kokh, S. Kashif Sadiq, Reggie Bosma, Indira Nederpelt, Laura H. Heitman, Elena Segala, Marta Amaral, Dong Guo, Dorothee Andres, Victoria Georgi, Leigh A. Stoddart, Steve Hill, Robert M. Cooke, Chris De Graaf, Rob Leurs, Matthias Frech, Rebecca C. Wade, Elizabeth Cunera Mariade Lange, Adriaan P. IJzerman, Anke Müller-Fahrnow, Gerhard F. Ecker

· DOI

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
Kd 3 8.78 9.69
k_off 3 - -
kon 3 - -

Assay Parameters

Parameter Type Value Units Text
assay_method assay_method - SPR
data_collection_rate data_collection_rate 10.0 Hz
expression_host expression_host - Insect cells
fitting_model fitting_model - 01:01
fitting_parameter_mass_transfer fitting_parameter_mass_transfer - global
fitting_parameter_offset fitting_parameter_offset - local
fitting_parameter_rmax fitting_parameter_rmax - global
immobilization_buffer_composition immobilization_buffer_composition - 20 mM Tris, 350 mM NaCl, pH 7.8
immobilization_chip_used immobilization_chip_used - CM5
immobilization_contact_time immobilization_contact_time 3600.0 s
immobilization_flow_rate immobilization_flow_rate 10.0 uL/min
immobilization_method immobilization_method - Antibody/Antigen coupling
immobilization_ph immobilization_ph 7.8
immobilization_temperature immobilization_temperature 18.0 degree C
instrument instrument - Biacore 3000
preparation_reference_surface preparation_reference_surface - The 1D4 antibody was immobilized via standard amino coupling on CM5 sensor surface to densities of 12-18 kRUs. Sensor surface is 10 min activated with 0.2 M EDC/ 0.05 M NHS mixture, antibody is contacted at 10 ug/ml in 10 mM sodium acetate buffer to reach required levels. Sensor surface is finally deactivated with 1 M ethanolamin, pH 8.0. A2A nanodsics are applied over 1D4 antibody surface to reach levels of 3000-6000 RUs.
protein_modification protein_modification - A2A-GFP-His10 embedded into nanodiscs containing C9-tag localized on the nanodisc' scaffold proteins.
protein_molecular_weight protein_molecular_weight 64.9 kDa
protein_mutation_type protein_mutation_type - mutated protein
reference1_fraction_active reference1_fraction_active 80.0 %

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL113142 ZM-241385 3 9.69
CHEMBL17127 SCH-58261 3 8.71
CHEMBL3981401 3 7.93

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL113142 ZM-241385 Kd = 0.206 nM 9.69
CHEMBL17127 SCH-58261 Kd = 1.973 nM 8.71
CHEMBL3981401 Kd = 11.66 nM 7.93
CHEMBL3981401 k_off = 0.02693 s-1 -
CHEMBL3981401 kon = 2190000.0 Ms-1 -
CHEMBL17127 SCH-58261 k_off = 0.004112 s-1 -
CHEMBL113142 ZM-241385 kon = 17676666.67 Ms-1 -
CHEMBL113142 ZM-241385 k_off = 0.003283 s-1 -
CHEMBL17127 SCH-58261 kon = 2110000.0 Ms-1 -