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 language | English (US) |
|---|---|
| Title of host publication | Proceedings of SPIE - The International Society for Optical Engineering |
| Editors | A.P. Savitsky, L.Y. Brovko, D.J. Bornhop, R. Raghavachari, S.I. Achilefu |
| Pages | 137-144 |
| Number of pages | 8 |
| Volume | 5329 |
| DOIs | |
| State | Published - 2004 |
| Externally published | Yes |
| Event | Genetically Engineered and Optical Probes for Biomedical Applications II - San Jose, CA, United States Duration: Jan 24 2004 → Jan 27 2004 |
Other
| Other | Genetically Engineered and Optical Probes for Biomedical Applications II |
|---|---|
| Country | United States |
| City | San Jose, CA |
| Period | 1/24/04 → 1/27/04 |
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Keywords
- Aequorin
- Bifunctional protein
- Bioluminescence
- Competitive immunoassay
- Endothelin-1
- Gene fusion
ASJC Scopus subject areas
- Electrical and Electronic Engineering
- Condensed Matter Physics
Cite this
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 proceeding › Conference contribution
}
TY - GEN
T1 - Aequorin fusion proteins as bioluminescent tracers for competitive immunoassays
AU - Mirasoli, Mara
AU - Michelini, Elisa
AU - Deo, Sapna K
AU - Dikici, Emre
AU - Roda, Aldo
AU - Daunert, Sylvia
PY - 2004
Y1 - 2004
N2 - 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.
AB - 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.
KW - Aequorin
KW - Bifunctional protein
KW - Bioluminescence
KW - Competitive immunoassay
KW - Endothelin-1
KW - Gene fusion
UR - http://www.scopus.com/inward/record.url?scp=7644226475&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=7644226475&partnerID=8YFLogxK
U2 - 10.1117/12.529194
DO - 10.1117/12.529194
M3 - Conference contribution
VL - 5329
SP - 137
EP - 144
BT - Proceedings of SPIE - The International Society for Optical Engineering
A2 - Savitsky, A.P.
A2 - Brovko, L.Y.
A2 - Bornhop, D.J.
A2 - Raghavachari, R.
A2 - Achilefu, S.I.
ER -