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Design and synthesis of immunoconjugates and development of competition inhibition enzyme-linked immunosorbent assay (CIEIA) for the detection of O-isopropyl methylphosphonofluoridate (sarin): An organophosphorous toxicant
[Display omitted] ► Production of polyclonal antibodies against organophosphorous toxicant sarin (GB). ► The haptens used in the present studies have not been studied so far. ► Antibodies showed negligible cross reactivity (CR) with other OP toxicants. ► CIEIA to distinguish GB agent in the presence...
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| Published in: | Journal of hazardous materials 2011-09, Vol.192 (3), p.1720-1728 |
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| container_issue | 3 |
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| container_title | Journal of hazardous materials |
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| creator | Sathe, Manisha Merwyn, S. Ghorpade, R. Agarwal, G.S. Rao, M.K. Rai, G.P. Kaushik, M.P. |
| description | [Display omitted]
► Production of polyclonal antibodies against organophosphorous toxicant sarin (GB). ► The haptens used in the present studies have not been studied so far. ► Antibodies showed negligible cross reactivity (CR) with other OP toxicants. ► CIEIA to distinguish GB agent in the presence of other OP toxicants or pesticide. ► Polyclonal antibodies for the selective detection of sarin.
Three haptens of the organophosphorus (OP) toxicant ‘sarin’ having different spacer arm were designed and synthesized. Haptens were conjugated with BSA (bovine serum albumin) and ovalbumin (OVA) for raising antibody and coating antigen. High antibody titer with higher specificity was obtained from 4-(4-(isopropoxy(methyl)phosphoryloxy)phenylamino)-4-oxobutanoic acid (hapten B) having reasonable long spacer arm. For the standard curve, an IC
50 (inhibitory concentration) of free antigen was found to be 0.415
μg
mL
−1 on the basis of indirect competitive ELISA. The study revealed that heterology in competition inhibition enzyme immunoassay (CIEIA) produced remarkable improvement in the sensitivity and specificity of the assay. Under the optimized conditions, the quantitative working range was found to be 0.19–1.56
μg
mL
−1 with a limit of detection (LOD) of 0.05
μg
mL
−1. The antibodies showed negligible cross reactivity (CR) with other OP toxicants and pesticides, which makes the assay suitable for the selective detection of sarin. |
| doi_str_mv | 10.1016/j.jhazmat.2011.07.011 |
| format | article |
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► Production of polyclonal antibodies against organophosphorous toxicant sarin (GB). ► The haptens used in the present studies have not been studied so far. ► Antibodies showed negligible cross reactivity (CR) with other OP toxicants. ► CIEIA to distinguish GB agent in the presence of other OP toxicants or pesticide. ► Polyclonal antibodies for the selective detection of sarin.
Three haptens of the organophosphorus (OP) toxicant ‘sarin’ having different spacer arm were designed and synthesized. Haptens were conjugated with BSA (bovine serum albumin) and ovalbumin (OVA) for raising antibody and coating antigen. High antibody titer with higher specificity was obtained from 4-(4-(isopropoxy(methyl)phosphoryloxy)phenylamino)-4-oxobutanoic acid (hapten B) having reasonable long spacer arm. For the standard curve, an IC
50 (inhibitory concentration) of free antigen was found to be 0.415
μg
mL
−1 on the basis of indirect competitive ELISA. The study revealed that heterology in competition inhibition enzyme immunoassay (CIEIA) produced remarkable improvement in the sensitivity and specificity of the assay. Under the optimized conditions, the quantitative working range was found to be 0.19–1.56
μg
mL
−1 with a limit of detection (LOD) of 0.05
μg
mL
−1. The antibodies showed negligible cross reactivity (CR) with other OP toxicants and pesticides, which makes the assay suitable for the selective detection of sarin.</description><identifier>ISSN: 0304-3894</identifier><identifier>EISSN: 1873-3336</identifier><identifier>DOI: 10.1016/j.jhazmat.2011.07.011</identifier><identifier>PMID: 21782327</identifier><identifier>CODEN: JHMAD9</identifier><language>eng</language><publisher>Kidlington: Elsevier B.V</publisher><subject>Animals ; Antibodies ; Antibodies - chemistry ; Applied sciences ; Bioconjugation ; bovine serum albumin ; Cattle ; Chemistry Techniques, Analytical ; cross reaction ; detection limit ; enzyme-linked immunosorbent assay ; Enzyme-Linked Immunosorbent Assay - methods ; Exact sciences and technology ; Female ; Gas Chromatography-Mass Spectrometry - methods ; Hapten ; haptens ; Haptens - chemistry ; Immunoconjugates - chemistry ; Immunoconjugates - immunology ; Inhibitory Concentration 50 ; Models, Chemical ; Organophosphorus Compounds - analysis ; ovalbumin ; Ovalbumin - chemistry ; pesticides ; Pollution ; Polyclonal antibody ; Protein ; Rabbits ; Reproducibility of Results ; Sarin ; Sarin - analysis ; Serum Albumin - chemistry ; Spectrophotometry, Ultraviolet - methods ; Temperature ; toxic substances ; Toxicant</subject><ispartof>Journal of hazardous materials, 2011-09, Vol.192 (3), p.1720-1728</ispartof><rights>2011 Elsevier B.V.</rights><rights>2015 INIST-CNRS</rights><rights>Copyright © 2011 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c450t-4d87db0e477849f707c70765c1396173278848e46db6fc293d0eb25f3b773fa83</citedby><cites>FETCH-LOGICAL-c450t-4d87db0e477849f707c70765c1396173278848e46db6fc293d0eb25f3b773fa83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24501502$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21782327$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sathe, Manisha</creatorcontrib><creatorcontrib>Merwyn, S.</creatorcontrib><creatorcontrib>Ghorpade, R.</creatorcontrib><creatorcontrib>Agarwal, G.S.</creatorcontrib><creatorcontrib>Rao, M.K.</creatorcontrib><creatorcontrib>Rai, G.P.</creatorcontrib><creatorcontrib>Kaushik, M.P.</creatorcontrib><title>Design and synthesis of immunoconjugates and development of competition inhibition enzyme-linked immunosorbent assay (CIEIA) for the detection of O-isopropyl methylphosphonofluoridate (sarin): An organophosphorous toxicant</title><title>Journal of hazardous materials</title><addtitle>J Hazard Mater</addtitle><description>[Display omitted]
► Production of polyclonal antibodies against organophosphorous toxicant sarin (GB). ► The haptens used in the present studies have not been studied so far. ► Antibodies showed negligible cross reactivity (CR) with other OP toxicants. ► CIEIA to distinguish GB agent in the presence of other OP toxicants or pesticide. ► Polyclonal antibodies for the selective detection of sarin.
Three haptens of the organophosphorus (OP) toxicant ‘sarin’ having different spacer arm were designed and synthesized. Haptens were conjugated with BSA (bovine serum albumin) and ovalbumin (OVA) for raising antibody and coating antigen. High antibody titer with higher specificity was obtained from 4-(4-(isopropoxy(methyl)phosphoryloxy)phenylamino)-4-oxobutanoic acid (hapten B) having reasonable long spacer arm. For the standard curve, an IC
50 (inhibitory concentration) of free antigen was found to be 0.415
μg
mL
−1 on the basis of indirect competitive ELISA. The study revealed that heterology in competition inhibition enzyme immunoassay (CIEIA) produced remarkable improvement in the sensitivity and specificity of the assay. Under the optimized conditions, the quantitative working range was found to be 0.19–1.56
μg
mL
−1 with a limit of detection (LOD) of 0.05
μg
mL
−1. The antibodies showed negligible cross reactivity (CR) with other OP toxicants and pesticides, which makes the assay suitable for the selective detection of sarin.</description><subject>Animals</subject><subject>Antibodies</subject><subject>Antibodies - chemistry</subject><subject>Applied sciences</subject><subject>Bioconjugation</subject><subject>bovine serum albumin</subject><subject>Cattle</subject><subject>Chemistry Techniques, Analytical</subject><subject>cross reaction</subject><subject>detection limit</subject><subject>enzyme-linked immunosorbent assay</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Exact sciences and technology</subject><subject>Female</subject><subject>Gas Chromatography-Mass Spectrometry - methods</subject><subject>Hapten</subject><subject>haptens</subject><subject>Haptens - chemistry</subject><subject>Immunoconjugates - chemistry</subject><subject>Immunoconjugates - immunology</subject><subject>Inhibitory Concentration 50</subject><subject>Models, Chemical</subject><subject>Organophosphorus Compounds - analysis</subject><subject>ovalbumin</subject><subject>Ovalbumin - chemistry</subject><subject>pesticides</subject><subject>Pollution</subject><subject>Polyclonal antibody</subject><subject>Protein</subject><subject>Rabbits</subject><subject>Reproducibility of Results</subject><subject>Sarin</subject><subject>Sarin - analysis</subject><subject>Serum Albumin - chemistry</subject><subject>Spectrophotometry, Ultraviolet - methods</subject><subject>Temperature</subject><subject>toxic substances</subject><subject>Toxicant</subject><issn>0304-3894</issn><issn>1873-3336</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><recordid>eNqFkcFu1DAURS0EotOBTwC8QbSLDHacxAkbNBoKjFSpC-jacpyXGQ-JHWynIv3YfgueToAlC-vZ8rnvXfsi9IqSFSW0eH9YHfbyvpdhlRJKV4SvYnmCFrTkLGGMFU_RgjCSJayssjN07v2BEEJ5nj1HZynlZcpSvkAPn8DrncHSNNhPJuzj0WPbYt33o7HKmsO4kwH8I9HAHXR26MGEI6NsP0DQQVuDtdnr-rQFcz_1kHTa_IBmbuStq48q6b2c8MVme7VdX-LWOhxHxr4B1KM2dr1JtLeDs8PU4R7CfuqGvfVxGdt2o3W6iX7whZdOm8sPeB1FbieNnSlnR4-D_aWVNOEFetbKzsPLuS7R7eer75uvyfXNl-1mfZ2oLCchyZqSNzWBjPMyq1pOuIqryBVlVUF5_KmyzErIiqYuWpVWrCFQp3nLas5ZK0u2RO9OfaPvnyP4IHrtFXSdNBD9iIqkrCQ0o5HMT6Ry1nsHrRic7qWbBCXimKw4iDlZcUxWEC5iibrX84Sx7qH5q_oTZQTezoD0Snatk0Zp_4-LD6V5tLFEb05cK62QOxeZ22_p8Y7QKuOsisTHEwHxx-40OOGVBqOg0S7GJBqr_2P2N0wk0_0</recordid><startdate>20110915</startdate><enddate>20110915</enddate><creator>Sathe, Manisha</creator><creator>Merwyn, S.</creator><creator>Ghorpade, R.</creator><creator>Agarwal, G.S.</creator><creator>Rao, M.K.</creator><creator>Rai, G.P.</creator><creator>Kaushik, M.P.</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7U7</scope><scope>C1K</scope></search><sort><creationdate>20110915</creationdate><title>Design and synthesis of immunoconjugates and development of competition inhibition enzyme-linked immunosorbent assay (CIEIA) for the detection of O-isopropyl methylphosphonofluoridate (sarin): An organophosphorous toxicant</title><author>Sathe, Manisha ; Merwyn, S. ; Ghorpade, R. ; Agarwal, G.S. ; Rao, M.K. ; Rai, G.P. ; Kaushik, M.P.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c450t-4d87db0e477849f707c70765c1396173278848e46db6fc293d0eb25f3b773fa83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Animals</topic><topic>Antibodies</topic><topic>Antibodies - chemistry</topic><topic>Applied sciences</topic><topic>Bioconjugation</topic><topic>bovine serum albumin</topic><topic>Cattle</topic><topic>Chemistry Techniques, Analytical</topic><topic>cross reaction</topic><topic>detection limit</topic><topic>enzyme-linked immunosorbent assay</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Exact sciences and technology</topic><topic>Female</topic><topic>Gas Chromatography-Mass Spectrometry - methods</topic><topic>Hapten</topic><topic>haptens</topic><topic>Haptens - chemistry</topic><topic>Immunoconjugates - chemistry</topic><topic>Immunoconjugates - immunology</topic><topic>Inhibitory Concentration 50</topic><topic>Models, Chemical</topic><topic>Organophosphorus Compounds - analysis</topic><topic>ovalbumin</topic><topic>Ovalbumin - chemistry</topic><topic>pesticides</topic><topic>Pollution</topic><topic>Polyclonal antibody</topic><topic>Protein</topic><topic>Rabbits</topic><topic>Reproducibility of Results</topic><topic>Sarin</topic><topic>Sarin - analysis</topic><topic>Serum Albumin - chemistry</topic><topic>Spectrophotometry, Ultraviolet - methods</topic><topic>Temperature</topic><topic>toxic substances</topic><topic>Toxicant</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sathe, Manisha</creatorcontrib><creatorcontrib>Merwyn, S.</creatorcontrib><creatorcontrib>Ghorpade, R.</creatorcontrib><creatorcontrib>Agarwal, G.S.</creatorcontrib><creatorcontrib>Rao, M.K.</creatorcontrib><creatorcontrib>Rai, G.P.</creatorcontrib><creatorcontrib>Kaushik, M.P.</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Toxicology Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Journal of hazardous materials</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sathe, Manisha</au><au>Merwyn, S.</au><au>Ghorpade, R.</au><au>Agarwal, G.S.</au><au>Rao, M.K.</au><au>Rai, G.P.</au><au>Kaushik, M.P.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Design and synthesis of immunoconjugates and development of competition inhibition enzyme-linked immunosorbent assay (CIEIA) for the detection of O-isopropyl methylphosphonofluoridate (sarin): An organophosphorous toxicant</atitle><jtitle>Journal of hazardous materials</jtitle><addtitle>J Hazard Mater</addtitle><date>2011-09-15</date><risdate>2011</risdate><volume>192</volume><issue>3</issue><spage>1720</spage><epage>1728</epage><pages>1720-1728</pages><issn>0304-3894</issn><eissn>1873-3336</eissn><coden>JHMAD9</coden><abstract>[Display omitted]
► Production of polyclonal antibodies against organophosphorous toxicant sarin (GB). ► The haptens used in the present studies have not been studied so far. ► Antibodies showed negligible cross reactivity (CR) with other OP toxicants. ► CIEIA to distinguish GB agent in the presence of other OP toxicants or pesticide. ► Polyclonal antibodies for the selective detection of sarin.
Three haptens of the organophosphorus (OP) toxicant ‘sarin’ having different spacer arm were designed and synthesized. Haptens were conjugated with BSA (bovine serum albumin) and ovalbumin (OVA) for raising antibody and coating antigen. High antibody titer with higher specificity was obtained from 4-(4-(isopropoxy(methyl)phosphoryloxy)phenylamino)-4-oxobutanoic acid (hapten B) having reasonable long spacer arm. For the standard curve, an IC
50 (inhibitory concentration) of free antigen was found to be 0.415
μg
mL
−1 on the basis of indirect competitive ELISA. The study revealed that heterology in competition inhibition enzyme immunoassay (CIEIA) produced remarkable improvement in the sensitivity and specificity of the assay. Under the optimized conditions, the quantitative working range was found to be 0.19–1.56
μg
mL
−1 with a limit of detection (LOD) of 0.05
μg
mL
−1. The antibodies showed negligible cross reactivity (CR) with other OP toxicants and pesticides, which makes the assay suitable for the selective detection of sarin.</abstract><cop>Kidlington</cop><pub>Elsevier B.V</pub><pmid>21782327</pmid><doi>10.1016/j.jhazmat.2011.07.011</doi><tpages>9</tpages></addata></record> |
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| ispartof | Journal of hazardous materials, 2011-09, Vol.192 (3), p.1720-1728 |
| issn | 0304-3894 1873-3336 |
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| source | ScienceDirect Journals |
| subjects | Animals Antibodies Antibodies - chemistry Applied sciences Bioconjugation bovine serum albumin Cattle Chemistry Techniques, Analytical cross reaction detection limit enzyme-linked immunosorbent assay Enzyme-Linked Immunosorbent Assay - methods Exact sciences and technology Female Gas Chromatography-Mass Spectrometry - methods Hapten haptens Haptens - chemistry Immunoconjugates - chemistry Immunoconjugates - immunology Inhibitory Concentration 50 Models, Chemical Organophosphorus Compounds - analysis ovalbumin Ovalbumin - chemistry pesticides Pollution Polyclonal antibody Protein Rabbits Reproducibility of Results Sarin Sarin - analysis Serum Albumin - chemistry Spectrophotometry, Ultraviolet - methods Temperature toxic substances Toxicant |
| title | Design and synthesis of immunoconjugates and development of competition inhibition enzyme-linked immunosorbent assay (CIEIA) for the detection of O-isopropyl methylphosphonofluoridate (sarin): An organophosphorous toxicant |
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