Element-tagged immunoassay with ICP-MS detection: evaluation and comparison to conventional immunoassays
Identifieur interne : 001216 ( Main/Exploration ); précédent : 001215; suivant : 001217Element-tagged immunoassay with ICP-MS detection: evaluation and comparison to conventional immunoassays
Auteurs : Eva Razumienko [Canada] ; Olga Ornatsky [Canada] ; Robert Kinach [Canada] ; Michael Milyavsky [Canada] ; Eric Lechman [Canada] ; Vladimir Baranov [Canada] ; Mitchell A. Winnik [Canada] ; Scott D. Tanner [Canada]Source :
- Journal of immunological methods [ 0022-1759 ] ; 2008.
Abstract
We have investigated the possibility of using element-tagged antibodies for protein detection and quantification in microplate format using Inductively Coupled Plasma Mass Spectrometry (ICP-MS), and compared the results to conventional immunoassays, such as Enzyme-Linked Immunosorbent Assay (ELISA) and Western blotting. The technique was further employed to detect low levels and measure DNA-binding activity of transcription factor p53 in leukemia cell lysates through its interaction with immobilized oligonucleotides and recognition by element-tagged antibodies. The advantages of ICP-MS detection for routine performance of immunoassays include increased sensitivity, wide dynamic range, minimal interference from complex matrices, and high throughput. Our approach advances the ICP-MS technology and demonstrates its applicability to proteomic studies through the use of antibodies directly labeled with polymer tags bearing multiple atoms of lanthanides. Development of this novel methodology will enable fast and quantitative identification of multiple analytes in a single well.
Url:
DOI: 10.1016/j.jim.2008.03.011
PubMed: 18456275
PubMed Central: 2583136
Affiliations:
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Winnik</name><affiliation wicri:level="4"><nlm:aff id="A1">Chemistry Department, University of Toronto, 80 St. George Street, Toronto, Ontario, Canada M5S 3H6</nlm:aff><country>Canada</country><wicri:regionArea>Chemistry Department, University of Toronto, 80 St. George Street, Toronto, Ontario</wicri:regionArea><orgName type="university">Université de Toronto</orgName><placeName><settlement type="city">Toronto</settlement><region type="state">Ontario</region></placeName></affiliation></author><author><name sortKey="Tanner, Scott D" sort="Tanner, Scott D" uniqKey="Tanner S" first="Scott D." last="Tanner">Scott D. Tanner</name><affiliation wicri:level="4"><nlm:aff id="A1">Chemistry Department, University of Toronto, 80 St. George Street, Toronto, Ontario, Canada M5S 3H6</nlm:aff><country>Canada</country><wicri:regionArea>Chemistry Department, University of Toronto, 80 St. George Street, Toronto, Ontario</wicri:regionArea><orgName type="university">Université de Toronto</orgName><placeName><settlement type="city">Toronto</settlement><region type="state">Ontario</region></placeName></affiliation></author></analytic><series><title level="j">Journal of immunological methods</title><idno type="ISSN">0022-1759</idno><imprint><date when="2008">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p id="P1">We have investigated the possibility of using element-tagged antibodies for protein detection and quantification in microplate format using Inductively Coupled Plasma Mass Spectrometry (ICP-MS), and compared the results to conventional immunoassays, such as Enzyme-Linked Immunosorbent Assay (ELISA) and Western blotting. The technique was further employed to detect low levels and measure DNA-binding activity of transcription factor p53 in leukemia cell lysates through its interaction with immobilized oligonucleotides and recognition by element-tagged antibodies. The advantages of ICP-MS detection for routine performance of immunoassays include increased sensitivity, wide dynamic range, minimal interference from complex matrices, and high throughput. Our approach advances the ICP-MS technology and demonstrates its applicability to proteomic studies through the use of antibodies directly labeled with polymer tags bearing multiple atoms of lanthanides. Development of this novel methodology will enable fast and quantitative identification of multiple analytes in a single well.</p></div></front></TEI><affiliations><list><country><li>Canada</li></country><region><li>Ontario</li></region><settlement><li>Toronto</li></settlement><orgName><li>Université de Toronto</li></orgName></list><tree><country name="Canada"><region name="Ontario"><name sortKey="Razumienko, Eva" sort="Razumienko, Eva" uniqKey="Razumienko E" first="Eva" last="Razumienko">Eva Razumienko</name></region><name sortKey="Baranov, Vladimir" sort="Baranov, Vladimir" uniqKey="Baranov V" first="Vladimir" last="Baranov">Vladimir Baranov</name><name sortKey="Kinach, Robert" sort="Kinach, Robert" uniqKey="Kinach R" first="Robert" last="Kinach">Robert Kinach</name><name sortKey="Lechman, Eric" sort="Lechman, Eric" uniqKey="Lechman E" first="Eric" last="Lechman">Eric Lechman</name><name sortKey="Milyavsky, Michael" sort="Milyavsky, Michael" uniqKey="Milyavsky M" first="Michael" last="Milyavsky">Michael Milyavsky</name><name sortKey="Ornatsky, Olga" sort="Ornatsky, Olga" uniqKey="Ornatsky O" first="Olga" last="Ornatsky">Olga Ornatsky</name><name sortKey="Tanner, Scott D" sort="Tanner, Scott D" uniqKey="Tanner S" first="Scott D." last="Tanner">Scott D. Tanner</name><name sortKey="Winnik, Mitchell A" sort="Winnik, Mitchell A" uniqKey="Winnik M" first="Mitchell A." last="Winnik">Mitchell A. Winnik</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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