title: Ultrasensitive ELISA for measurement of human cytokine responses in primary culture.
abstract: ELISAs offer excellent specificity and, once fully optimized, sensitivity that rivals that of bioassays. The major variables that need to be experimentally determined when developing an ELISA are the optimal number of fresh cells required per well, the optimal antigen concentrations for stimulation, period of culture, and the anticipated intensity of the response. In this chapter, we review the major factors to be considered in the development and application of ultrasensitive ELISAs to the analysis of human immune responses. We specify the conditions we have found to be optimal for quantifying a number of cytokines of demonstrated relevance to human immune regulation and discuss the major pitfalls inherent in this approach.
authors.affiliation: Department of Immunology, University of Manitoba, Winnipeg, Canada.
authors.forename: William P
authors.initials: WP
authors.lastname: Stefura
authors.forename: J Darren
authors.initials: JD
authors.lastname: Campbell
authors.forename: Renée
authors.initials: R
authors.lastname: Douville
authors.forename: Monique J
authors.initials: MJ
authors.lastname: Stinson
authors.forename: F Estelle
authors.initials: FE
authors.lastname: Simons
authors.forename: Allan B
authors.initials: AB
authors.lastname: Becker
authors.forename: Kent T
authors.initials: KT
authors.lastname: HayGlass
mesh_terms: D002478:Cells, Cultured; D016207:Cytokines / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000191:economics / Q000379:methods* / Q000592:standards; D006801:Humans; D008163:Luminescent Measurements; D012015:Reference Standards; D012680:Sensitivity and Specificity
chemical_list: D016207:Cytokines