. Author manuscript; available in PMC: 2023 Sep 15.

Published in final edited form as: J Immunol Methods. 2015 May 7;424:20–27. doi: 10.1016/j.jim.2015.04.017

Table 1.

Comparison of “Ideal” ligand binding assay system proposed by Spriggs et al.,² and the automated Simoa instrument presented here.

Platform Attribute	Measure of Success (Sprigg et al. AAPS J., 2012)²	Automated Simoa performance
Sensitivity	Capable of quantifying low analyte concentrations of drug and biomarkers in a variety of matrices	1000-fold more sensitive than standard immunoassays^a^,3,8 fg/mL sensitivity^a
Dynamic Range	Greater than 3 log range	>4 log range^a^,11
Precision	Less than 2% variability for the instrument signal of internal standard	4.2% variability in AEB^a
Ruggedness	Consistent performance under varied laboratory conditions	Full automation has the potential to minimize lab-to-lab variability^c
Ruggedness	Tolerance to biological interferences	Standard immunoassay approaches to reduce interferences⁸
Total Assay time	Results in 1 h or less	Time to first result = 83 min^a (45 min also demonstrated)^b
Multiplexing	Should have capabilities	6-plex immunoassay demonstrated;^a 10-plex decoding demonstrated^b
Multiplexing	Minimal cross-talk due to detection mechanisms	Optical cross-talk <0.1%¹⁴
Flexibility/throughput	Automation compatible	Fully automated^a
	Capable of assay miniaturization (e.g., 384+ well plates or other substrate)	96-well plates and primary tubes as sample input^a
	Utilization of various solid supports	Standard immobilization chemistries compatible with beads^a
	Ability to run in low and high throughput environments	Yes^a
	Possible to use throughout the life cycle of drug development	Yes. Available for researchers, contract-research organizations, and under development for regulatory approval^a
Multi-modality	Measure wide variety of therapeutics including proteins, peptides, antibodies, etc.	Proteins,^a antibodies,^b DNA¹⁸ demonstrated
Life Cycle Support	Multiple sources for reagent availability	Yes. Many commercial sources of antibody pairs.⁸
Life Cycle Support	Ability to label reagents in-house	Yes. Open platform via “homebrew” kits.^a

This work.

Unpublished work.

Not tested in this work.