title: Comparison of four distinct detection platforms using multiple ligand binding assay formats.
abstract: Several detection platforms are available for ligand binding assays (LBA), each claiming superiority in sensitivity and dynamic range. However, little information exists in the literature directly comparing the various LBA platforms for quantitation. We have tested four common platforms to evaluate and compare the interchangeability of detection platforms by comparing sensitivity and dynamic range to a colorimetric LBA. The detection platforms compared are: colorimetric, chemiluminescence, time-resolved fluorescence (TRF) and electrochemiluminescence (ECL). Five different LBA protocols were tested with each of the detection endpoints. The assay protocols include the following ligand binding assay formats: direct binding, sandwich ELISA, competitive and cell based ELISA. We found that no detection platform consistently performed better than all the others and it was not possible to predict which platform would perform best for a given assay protocol. We also found surprising differences in assays (plate coating efficiency, low signal) which add to difficulty in choosing the best platform ad hoc. We propose here that in developing new assay protocols for detection of biotherapeutic agents, multiple detection platforms should be tested in order to forward the best assays possible and for the right reasons.
authors.affiliation: Pfizer, Inc, 558 Eastern Point Road Groton, CT 06340, United States.
authors.forename: Catherine I
authors.initials: CI
authors.lastname: Soderstrom
authors.forename: Franklin P
authors.initials: FP
authors.lastname: Spriggs
authors.forename: Wei
authors.initials: W
authors.lastname: Song
authors.forename: Sarah
authors.initials: S
authors.lastname: Burrell
mesh_terms: D030301:Activin Receptors, Type II / Q000032:analysis; D000818:Animals; D003124:Colorimetry / Q000379:methods / Q000706:statistics & numerical data; D055664:Electrochemical Techniques / Q000379:methods / Q000706:statistics & numerical data; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000706:statistics & numerical data; D005934:Glucagon / Q000032:analysis; D000882:Haplorhini; D006801:Humans; D007118:Immunoassay / Q000379:methods* / Q000706:statistics & numerical data; D008024:Ligands; D008163:Luminescent Measurements / Q000379:methods / Q000706:statistics & numerical data; D017526:Receptor, IGF Type 1 / Q000032:analysis; D011994:Recombinant Proteins / Q000032:analysis; D012680:Sensitivity and Specificity
chemical_list: D008024:Ligands; D011994:Recombinant Proteins; D005934:Glucagon; D017526:Receptor, IGF Type 1; C425364:ACVRL1 protein, human; D030301:Activin Receptors, Type II