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Enzyme linked immuno mass spectrometric assay (ELIMSA).
J Proteomics. 2014 Jan 16; 96:343-52.JP

Abstract

A new technology termed ELIMSA combines the specificity and enzymatic amplification of Enzyme Linked Immunosorbent assay (ELISA) with the sensitivity and flexibility of mass spectrometry (MS). At present, substrates for the reporter enzymes horseradish peroxidase (HRP) or alkaline phosphatase (AP) yield colored, fluorescent or luminescent products. The central concept of ELIMSA is that the reporter enzymes HRP and AP yield products that ionize efficiently with a high signal to noise ratio that can be measured by mass spectrometry. The reporter enzymes HRP or AP may be covalently attached to a specific detection probe such as a protein or an antibody to bind their target analyte and then catalyze the rapid production of ionizable, small-molecules. The use of mass spectrometry to measure small molecule products may commonly reach femto to picomol amounts on the column with high signal to noise ratio. Mass spectrometry combined with the enzyme amplification in ELISA provides absolute sensitivity to detect attomol of PSA and was comparable to, or more sensitive, than radio immune assays and electrochemical detectors but with only existing reagents and equipment. ELIMSA permits monitoring of multiple substrates and products and provides comparison to absolute standards.

BIOLOGICAL SIGNIFICANCE

There is an urgent need to detect and quantify low abundance proteins such as hormones, chemokines, cytokines, and others that exist at attomolar concentrations under physiological conditions by ELIMSA. A sensitive method for the quantification of immunological assays is obtained by applying mass spectrometry to detect the products of the alkaline phosphatase (AP) and horseradish peroxidase (HRP) enzyme reactions. There are many molecules from human subjects or micro-organisms that are of great importance to medicine, industry, nutrition or the environment that need to be repeatedly analyzed and are often near to, or beyond, the edge of existing analytical technology. The presence of molecules in biological samples, industrial products or the environment may be detected by probes that bind to the target analyte. Combining the reporter enzymes from ELISA with sensitive liquid chromatography (LC), electrospray ionization (ESI) and tandem mass spectrometry (MS/MS) will permit the sensitive detection and quantification of the molecular probes by Enzyme Linked Immuno Mass Spectrometric Assay (ELIMSA). The flexibility and sensitivity of mass spectrometry to measure large numbers of compounds simultaneously should permit the quantification of multiple ELIMSA reactions at separate mass-to-charge (m/z) ratios. Hence ELIMSA and it variants should permit the rapid and simple detection and quantification of many molecules over the complete range of biologically important concentrations without the use of radiolabels using only existing antibodies, reagents and instruments. Antibodies coupled to reporter enzymes that are widely used in biomedical and environmental applications can now be detected and quantified using ultra sensitive mass spectrometry to create a sensitive and flexible ELIMSA system. Absolute standards of analytes or enzyme product may serve as a reference.

Authors+Show Affiliations

Department of Chemistry and Biology, Faculty of Science, Ryerson University, Toronto, Canada.Department of Chemistry and Biology, Faculty of Science, Ryerson University, Toronto, Canada.Department of Chemistry and Biology, Faculty of Science, Ryerson University, Toronto, Canada. Electronic address: 4marshal@ryerson.ca.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24316356

Citation

Florentinus-Mefailoski, Angelique, et al. "Enzyme Linked Immuno Mass Spectrometric Assay (ELIMSA)." Journal of Proteomics, vol. 96, 2014, pp. 343-52.
Florentinus-Mefailoski A, Safi F, Marshall JG. Enzyme linked immuno mass spectrometric assay (ELIMSA). J Proteomics. 2014;96:343-52.
Florentinus-Mefailoski, A., Safi, F., & Marshall, J. G. (2014). Enzyme linked immuno mass spectrometric assay (ELIMSA). Journal of Proteomics, 96, 343-52. https://doi.org/10.1016/j.jprot.2013.11.022
Florentinus-Mefailoski A, Safi F, Marshall JG. Enzyme Linked Immuno Mass Spectrometric Assay (ELIMSA). J Proteomics. 2014 Jan 16;96:343-52. PubMed PMID: 24316356.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enzyme linked immuno mass spectrometric assay (ELIMSA). AU - Florentinus-Mefailoski,Angelique, AU - Safi,Frozan, AU - Marshall,John G, Y1 - 2013/12/04/ PY - 2013/08/10/received PY - 2013/11/16/revised PY - 2013/11/25/accepted PY - 2013/12/10/entrez PY - 2013/12/10/pubmed PY - 2014/9/23/medline KW - AP KW - AR KW - ASMX KW - ASMX naphthol KW - ASMX naphthol phosphate KW - ASMXP KW - Alkaline Phosphatase KW - Amplex red KW - Amplex red (10-acetyl-3,7-dihydroxyphenoxazine) KW - CID KW - DF KW - ECL KW - ELIMSA KW - ELISA KW - Enzyme Linked Immuno Mass Spectrometric Assay KW - Enzyme Linked Immuno-Sorbent Assay KW - H(2)O(2) KW - HRP KW - Horseradish peroxidase KW - MS KW - MS/MS KW - Mass spectrometry KW - Naphthol ASMX phosphate KW - SA KW - SIM KW - SRM KW - alkaline phosphatase KW - collision-induced dissociation KW - dilution factor KW - enhanced chemiluminescence KW - horseradish peroxidase KW - hydrogen peroxide KW - i KW - intensity KW - mass spectrometry KW - single ion monitoring KW - single reaction monitoring KW - streptavidin KW - tandem mass spectrometry SP - 343 EP - 52 JF - Journal of proteomics JO - J Proteomics VL - 96 N2 - UNLABELLED: A new technology termed ELIMSA combines the specificity and enzymatic amplification of Enzyme Linked Immunosorbent assay (ELISA) with the sensitivity and flexibility of mass spectrometry (MS). At present, substrates for the reporter enzymes horseradish peroxidase (HRP) or alkaline phosphatase (AP) yield colored, fluorescent or luminescent products. The central concept of ELIMSA is that the reporter enzymes HRP and AP yield products that ionize efficiently with a high signal to noise ratio that can be measured by mass spectrometry. The reporter enzymes HRP or AP may be covalently attached to a specific detection probe such as a protein or an antibody to bind their target analyte and then catalyze the rapid production of ionizable, small-molecules. The use of mass spectrometry to measure small molecule products may commonly reach femto to picomol amounts on the column with high signal to noise ratio. Mass spectrometry combined with the enzyme amplification in ELISA provides absolute sensitivity to detect attomol of PSA and was comparable to, or more sensitive, than radio immune assays and electrochemical detectors but with only existing reagents and equipment. ELIMSA permits monitoring of multiple substrates and products and provides comparison to absolute standards. BIOLOGICAL SIGNIFICANCE: There is an urgent need to detect and quantify low abundance proteins such as hormones, chemokines, cytokines, and others that exist at attomolar concentrations under physiological conditions by ELIMSA. A sensitive method for the quantification of immunological assays is obtained by applying mass spectrometry to detect the products of the alkaline phosphatase (AP) and horseradish peroxidase (HRP) enzyme reactions. There are many molecules from human subjects or micro-organisms that are of great importance to medicine, industry, nutrition or the environment that need to be repeatedly analyzed and are often near to, or beyond, the edge of existing analytical technology. The presence of molecules in biological samples, industrial products or the environment may be detected by probes that bind to the target analyte. Combining the reporter enzymes from ELISA with sensitive liquid chromatography (LC), electrospray ionization (ESI) and tandem mass spectrometry (MS/MS) will permit the sensitive detection and quantification of the molecular probes by Enzyme Linked Immuno Mass Spectrometric Assay (ELIMSA). The flexibility and sensitivity of mass spectrometry to measure large numbers of compounds simultaneously should permit the quantification of multiple ELIMSA reactions at separate mass-to-charge (m/z) ratios. Hence ELIMSA and it variants should permit the rapid and simple detection and quantification of many molecules over the complete range of biologically important concentrations without the use of radiolabels using only existing antibodies, reagents and instruments. Antibodies coupled to reporter enzymes that are widely used in biomedical and environmental applications can now be detected and quantified using ultra sensitive mass spectrometry to create a sensitive and flexible ELIMSA system. Absolute standards of analytes or enzyme product may serve as a reference. SN - 1876-7737 UR - https://www.unboundmedicine.com/medline/citation/24316356/Enzyme_linked_immuno_mass_spectrometric_assay__ELIMSA__ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1874-3919(13)00629-5 DB - PRIME DP - Unbound Medicine ER -