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Review

Immunoassay Methods

Karen L. Cox  1 , Viswanath Devanarayan  2 , Aidas Kriauciunas  1 , Joseph Manetta  1 , Chahrzad Montrose  1 , Sitta Sittampalam  3
Sarine Markossian  1 , Abigail Grossman  1 , Hannah Baskir  1 , Michelle Arkin  2 , Douglas Auld  3 , Christopher Austin  4 , Jonathan Baell  5 , Kyle Brimacombe  1 , Thomas D.Y. Chung  6 , Nathan P. Coussens  7 , Jayme L. Dahlin  8 , Viswanath Devanarayan  9 , Timothy L. Foley  10 , Marcie Glicksman  11 , Kirill Gorshkov  12 , Stefan Grotegut  13 , Matthew D. Hall  1 , Samuel Hoare  14 , James Inglese  1 , Philip W. Iversen  15 , Madhu Lal-Nag  16 , Zhuyin Li  12 , Jason R. Manro  13 , James McGee  17 , Allison Norvil  13 , Mackenzie Pearson  13 , Terry Riss  18 , Peter Saradjian  19 , G. Sitta Sittampalam  20 , Michael A. Tarselli  21 , O. Joseph Trask Jr.  22 , Jeffrey R. Weidner  23 , Mary Jo Wildey  24 , Kelli Wilson  1 , Menghang Xia  1 , Xin Xu  1
, editors.
In: Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004.
[updated ].
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Review

Immunoassay Methods

Karen L. Cox et al.
Free Books & Documents

Excerpt

Immunoassays are used to quantify molecules of biological interest based on the specificity and selectivity of antibody reagents generated. In HTS and lead optimization projects, assays are designed to detect molecules that are produced intracellularly or secreted in response to compounds screened. This chapter describes the basics of designing and implementing robust, automation friendly immunoassays for HTS, modes of immunoassay formats (competitive and sandwich), instrumentation, reagent selection, experimental design and detailed data analysis concepts. The importance of an appropriate curve-fitting model for calibration curves used for quantification is also addressed in detail. This is an excellent primer for beginners as well as for experienced investigators.

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References

    1. Swartzman, et al. A Homogeneous Multiplexed Immunoassay for High-Throughput Screening Using Fluorometric Microvolume Assay Technology. Anal. Biochem. 1999;271(2):143–151. - PubMed
    1. Stockwell, et al. High-throughput screening of small molecules in miniaturized mammalian cell-based assays involving post-translational modifications. Chem. Biol. 1999;6(2):71–83. - PubMed
    1. Martens, et al. A generic particle-based nonradioactive homogeneous multiplex method for high-throughput screening using microvolume fluorimetry. Anal. Biochem. 1999;273(1):20–31. - PubMed
    1. Zuck, et al. Ligand-receptor binding measured by laser-scanning imaging. Proc. Natl. Acad. Sci. 1999;96(20):11122–11127. - PMC - PubMed
    1. Kitamura, et al. A fluorescence sandwich ELISA for detecting soluble and cell-associated human interleukin-2. J. Immunol. Methods. 1989;121(2):281–288. - PubMed

Additional References

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    1. http://www.piercenet.com/method/elisa-development-optimization
    1. http://www.piercenet.com/method/overview-elisa#elisaprobes
    1. http://www.piercenet.com/method/overview-detection-probes

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