Aequorin fusion proteins as bioluminescent tracers for competitive immunoassays

Mara Mirasoli, Elisa Michelini, Sapna K Deo, Emre Dikici, Aldo Roda, Sylvia Daunert

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Citation (Scopus)

Abstract

The use of bio- and chemiluminescence for the development of quantitative binding assays offers undoubted advantages over other detection systems, such as spectrophotometry, fluorescence, or radioactivity. Indeed, bio- and chemiluminescence detection provides similar, or even better, sensitivity and detectability than radioisotopes, while avoiding the problems of health hazards, waste disposal, and instability associated with the use of radioisotopes. Among bioluminescent labels, the calcium-activated photoprotein aequorin, originally isolated from Aequorea victoria and today available as a recombinant product, is characterized by very high detectability, down to attomole levels. It has been used as a bioluminescent label for developing a variety of highly sensitive immunoassays, using various analyte-aequorin conjugation strategies. When the analyte is a protein or a peptide, genetic engineering techniques can be used to produce protein fusions where the analyte is in-frame fused with aequorin, thus producing homogeneous one-to-one conjugation products, available in virtually unlimited amount. Various assays were developed using this strategy: a short review of the most interesting applications is presented, as well as the cloning, purification and initial characterization of an endothelin-1-aequorin conjugate suitable for developing a competitive immunoassay for endothelin-1, a potent vasoconstrictor peptide, involved in hypertension.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
EditorsA.P. Savitsky, L.Y. Brovko, D.J. Bornhop, R. Raghavachari, S.I. Achilefu
Pages137-144
Number of pages8
Volume5329
DOIs
StatePublished - 2004
Externally publishedYes
EventGenetically Engineered and Optical Probes for Biomedical Applications II - San Jose, CA, United States
Duration: Jan 24 2004Jan 27 2004

Other

OtherGenetically Engineered and Optical Probes for Biomedical Applications II
CountryUnited States
CitySan Jose, CA
Period1/24/041/27/04

Fingerprint

immunoassay
Chemiluminescence
chemiluminescence
conjugation
Radioisotopes
Peptides
peptides
tracers
Labels
Assays
Fusion reactions
fusion
proteins
Proteins
waste disposal
hypertension
Genetic engineering
Health hazards
Cloning
Radioactivity

Keywords

  • Aequorin
  • Bifunctional protein
  • Bioluminescence
  • Competitive immunoassay
  • Endothelin-1
  • Gene fusion

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

Cite this

Mirasoli, M., Michelini, E., Deo, S. K., Dikici, E., Roda, A., & Daunert, S. (2004). Aequorin fusion proteins as bioluminescent tracers for competitive immunoassays. In A. P. Savitsky, L. Y. Brovko, D. J. Bornhop, R. Raghavachari, & S. I. Achilefu (Eds.), Proceedings of SPIE - The International Society for Optical Engineering (Vol. 5329, pp. 137-144) https://doi.org/10.1117/12.529194

Aequorin fusion proteins as bioluminescent tracers for competitive immunoassays. / Mirasoli, Mara; Michelini, Elisa; Deo, Sapna K; Dikici, Emre; Roda, Aldo; Daunert, Sylvia.

Proceedings of SPIE - The International Society for Optical Engineering. ed. / A.P. Savitsky; L.Y. Brovko; D.J. Bornhop; R. Raghavachari; S.I. Achilefu. Vol. 5329 2004. p. 137-144.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Mirasoli, M, Michelini, E, Deo, SK, Dikici, E, Roda, A & Daunert, S 2004, Aequorin fusion proteins as bioluminescent tracers for competitive immunoassays. in AP Savitsky, LY Brovko, DJ Bornhop, R Raghavachari & SI Achilefu (eds), Proceedings of SPIE - The International Society for Optical Engineering. vol. 5329, pp. 137-144, Genetically Engineered and Optical Probes for Biomedical Applications II, San Jose, CA, United States, 1/24/04. https://doi.org/10.1117/12.529194
Mirasoli M, Michelini E, Deo SK, Dikici E, Roda A, Daunert S. Aequorin fusion proteins as bioluminescent tracers for competitive immunoassays. In Savitsky AP, Brovko LY, Bornhop DJ, Raghavachari R, Achilefu SI, editors, Proceedings of SPIE - The International Society for Optical Engineering. Vol. 5329. 2004. p. 137-144 https://doi.org/10.1117/12.529194
Mirasoli, Mara ; Michelini, Elisa ; Deo, Sapna K ; Dikici, Emre ; Roda, Aldo ; Daunert, Sylvia. / Aequorin fusion proteins as bioluminescent tracers for competitive immunoassays. Proceedings of SPIE - The International Society for Optical Engineering. editor / A.P. Savitsky ; L.Y. Brovko ; D.J. Bornhop ; R. Raghavachari ; S.I. Achilefu. Vol. 5329 2004. pp. 137-144
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