A Microarray Immunoassay for Serum Thyrotropin and Thyroglobulin Using Antibodies Immobilized on Track-Etched Membranes
Serum thyroglobulin (Tg) and thyroid stimulating hormone (TSH) measurements have evolved as important analytes for monitoring the prognosis of patients with differentiated thyroid cancer, post-thyroidectomy. Individual analyte immunoassay is the current practice in clinical pathology, but the simult...
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| Veröffentlicht in: | Indian journal of clinical biochemistry 2017-06, Vol.32 (2), p.193-199 |
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| creator | Jain, Bharti Kumarasamy, J. Gholve, Chandrakala Kulkarni, Savita Rajan, M. G. R. |
| description | Serum thyroglobulin (Tg) and thyroid stimulating hormone (TSH) measurements have evolved as important analytes for monitoring the prognosis of patients with differentiated thyroid cancer, post-thyroidectomy. Individual analyte immunoassay is the current practice in clinical pathology, but the simultaneous assay for all relevant analytes for a given disease, can reduce assay costs, improve patient compliance and give the clinician more information for an unequivocal diagnosis. Microarray immunoassay (MI) can achieve this goal and, hence, we have developed and validated a immuno-radiometric MI for quantitation of serum TSH and Tg by using highly micro-porous polycarbonate (PC) track-etched membranes (TEM) to immobilize the monoclonal anti-TSH and polyclonal anti-Tg antibodies in ~1 mm diameter spots. Non-competitive immunoassays were performed using mixture of
125
I labeled monoclonal anti-TSH and anti-Tg antibodies. Phosphorimager was used to quantify the bound radioactivity. TSH and Tg were detected with detection limit of 0.07 µIU/ml and 0.13 ng/ml respectively, which is lower than the clinically required cut-off level. The assay showed: acceptable intra-assay precision within 20 % and recovery in the range of 76–111.2 %. MI compared well with the established immunoradiometric assay (IRMA) with r = 0.98,
p
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| doi_str_mv | 10.1007/s12291-016-0589-2 |
| format | Article |
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125
I labeled monoclonal anti-TSH and anti-Tg antibodies. Phosphorimager was used to quantify the bound radioactivity. TSH and Tg were detected with detection limit of 0.07 µIU/ml and 0.13 ng/ml respectively, which is lower than the clinically required cut-off level. The assay showed: acceptable intra-assay precision within 20 % and recovery in the range of 76–111.2 %. MI compared well with the established immunoradiometric assay (IRMA) with r = 0.98,
p
< 0.01 (n = 41). No cross-reactivity was seen between the immobilized antibodies. Although two hormones are addressed in this report, MI using PC TEM and isotopic/non-isotopic tracers has the potential for highly automated multiplexed analysis.</description><identifier>ISSN: 0970-1915</identifier><identifier>EISSN: 0974-0422</identifier><identifier>DOI: 10.1007/s12291-016-0589-2</identifier><identifier>PMID: 28428694</identifier><language>eng</language><publisher>New Delhi: Springer India</publisher><subject>Antibodies ; Biochemistry ; Biomedical and Life Sciences ; Chemistry/Food Science ; Hormones ; Immunoassay ; Life Sciences ; Microbiology ; Original ; Original Article ; Pathology ; Patient compliance ; Polyclonal antibodies ; Prognosis ; Thyroid cancer ; Thyroid gland ; Thyrotropin ; Viral antibodies</subject><ispartof>Indian journal of clinical biochemistry, 2017-06, Vol.32 (2), p.193-199</ispartof><rights>Association of Clinical Biochemists of India 2016</rights><rights>COPYRIGHT 2017 Springer</rights><rights>Indian Journal of Clinical Biochemistry is a copyright of Springer, 2017.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c604t-aa2177b68a103da448633752b8dc352d3834091f70ca55e6ee84a6d6773e33963</citedby><cites>FETCH-LOGICAL-c604t-aa2177b68a103da448633752b8dc352d3834091f70ca55e6ee84a6d6773e33963</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5382074/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5382074/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,41464,42533,51294,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28428694$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jain, Bharti</creatorcontrib><creatorcontrib>Kumarasamy, J.</creatorcontrib><creatorcontrib>Gholve, Chandrakala</creatorcontrib><creatorcontrib>Kulkarni, Savita</creatorcontrib><creatorcontrib>Rajan, M. G. R.</creatorcontrib><title>A Microarray Immunoassay for Serum Thyrotropin and Thyroglobulin Using Antibodies Immobilized on Track-Etched Membranes</title><title>Indian journal of clinical biochemistry</title><addtitle>Ind J Clin Biochem</addtitle><addtitle>Indian J Clin Biochem</addtitle><description>Serum thyroglobulin (Tg) and thyroid stimulating hormone (TSH) measurements have evolved as important analytes for monitoring the prognosis of patients with differentiated thyroid cancer, post-thyroidectomy. Individual analyte immunoassay is the current practice in clinical pathology, but the simultaneous assay for all relevant analytes for a given disease, can reduce assay costs, improve patient compliance and give the clinician more information for an unequivocal diagnosis. Microarray immunoassay (MI) can achieve this goal and, hence, we have developed and validated a immuno-radiometric MI for quantitation of serum TSH and Tg by using highly micro-porous polycarbonate (PC) track-etched membranes (TEM) to immobilize the monoclonal anti-TSH and polyclonal anti-Tg antibodies in ~1 mm diameter spots. Non-competitive immunoassays were performed using mixture of
125
I labeled monoclonal anti-TSH and anti-Tg antibodies. Phosphorimager was used to quantify the bound radioactivity. TSH and Tg were detected with detection limit of 0.07 µIU/ml and 0.13 ng/ml respectively, which is lower than the clinically required cut-off level. The assay showed: acceptable intra-assay precision within 20 % and recovery in the range of 76–111.2 %. MI compared well with the established immunoradiometric assay (IRMA) with r = 0.98,
p
< 0.01 (n = 41). No cross-reactivity was seen between the immobilized antibodies. Although two hormones are addressed in this report, MI using PC TEM and isotopic/non-isotopic tracers has the potential for highly automated multiplexed analysis.</description><subject>Antibodies</subject><subject>Biochemistry</subject><subject>Biomedical and Life Sciences</subject><subject>Chemistry/Food Science</subject><subject>Hormones</subject><subject>Immunoassay</subject><subject>Life Sciences</subject><subject>Microbiology</subject><subject>Original</subject><subject>Original Article</subject><subject>Pathology</subject><subject>Patient compliance</subject><subject>Polyclonal antibodies</subject><subject>Prognosis</subject><subject>Thyroid cancer</subject><subject>Thyroid gland</subject><subject>Thyrotropin</subject><subject>Viral antibodies</subject><issn>0970-1915</issn><issn>0974-0422</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>BENPR</sourceid><recordid>eNqNkk1v1DAQhiMEoh_wA7igSFzaQ4q_YjsXpFVVYKVWSHR7thzHybok9mInwPLrmSWldBFIyAePPc-81ozfLHuB0RlGSLxOmJAKFwjzApWyKsij7BBVghWIEfL4Z4wKXOHyIDtK6RYhyhDDT7MDIhmRvGKH2ddFfuVMDDpGvc2XwzD5oFOCuA0xv7ZxGvLVehvDGMPG-Vz7Zj53fainHm5ukvNdvvCjq0PjbNqJhNr17rtt8uDzVdTmU3ExmjWcr-xQR-1tepY9aXWf7PO7_Ti7eXuxOn9fXH54tzxfXBaGIzYWWhMsRM2lxog2mjHJKRUlqWVjaEkaKqGlCrcCGV2WllsrmeYNF4JaSitOj7M3s-5mqgfbGOvHqHu1iW7QcauCdmo_491adeGLKqkkSDAQOLkTiOHzZNOoBpeM7XvoIkxJYVlhKWHS9L9QTKUsMaCv_kBvwxQ9TAIoyRCXFeO_qU73Vjnfwi9osxNVC4EZfCaSu2fP_kLBauzgTPC2dXC_V3C6VwDMaL-NnZ5SUsvrj_ssnlnwSErRtvejw0jtTKhmEyowodqZUBGoeflw5vcVv1wHAJmBBCnf2fig-3-q_gBGk-V-</recordid><startdate>20170601</startdate><enddate>20170601</enddate><creator>Jain, Bharti</creator><creator>Kumarasamy, J.</creator><creator>Gholve, Chandrakala</creator><creator>Kulkarni, Savita</creator><creator>Rajan, M. 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R.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c604t-aa2177b68a103da448633752b8dc352d3834091f70ca55e6ee84a6d6773e33963</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Antibodies</topic><topic>Biochemistry</topic><topic>Biomedical and Life Sciences</topic><topic>Chemistry/Food Science</topic><topic>Hormones</topic><topic>Immunoassay</topic><topic>Life Sciences</topic><topic>Microbiology</topic><topic>Original</topic><topic>Original Article</topic><topic>Pathology</topic><topic>Patient compliance</topic><topic>Polyclonal antibodies</topic><topic>Prognosis</topic><topic>Thyroid cancer</topic><topic>Thyroid gland</topic><topic>Thyrotropin</topic><topic>Viral antibodies</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jain, Bharti</creatorcontrib><creatorcontrib>Kumarasamy, J.</creatorcontrib><creatorcontrib>Gholve, Chandrakala</creatorcontrib><creatorcontrib>Kulkarni, Savita</creatorcontrib><creatorcontrib>Rajan, M. 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G. R.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Microarray Immunoassay for Serum Thyrotropin and Thyroglobulin Using Antibodies Immobilized on Track-Etched Membranes</atitle><jtitle>Indian journal of clinical biochemistry</jtitle><stitle>Ind J Clin Biochem</stitle><addtitle>Indian J Clin Biochem</addtitle><date>2017-06-01</date><risdate>2017</risdate><volume>32</volume><issue>2</issue><spage>193</spage><epage>199</epage><pages>193-199</pages><issn>0970-1915</issn><eissn>0974-0422</eissn><abstract>Serum thyroglobulin (Tg) and thyroid stimulating hormone (TSH) measurements have evolved as important analytes for monitoring the prognosis of patients with differentiated thyroid cancer, post-thyroidectomy. Individual analyte immunoassay is the current practice in clinical pathology, but the simultaneous assay for all relevant analytes for a given disease, can reduce assay costs, improve patient compliance and give the clinician more information for an unequivocal diagnosis. Microarray immunoassay (MI) can achieve this goal and, hence, we have developed and validated a immuno-radiometric MI for quantitation of serum TSH and Tg by using highly micro-porous polycarbonate (PC) track-etched membranes (TEM) to immobilize the monoclonal anti-TSH and polyclonal anti-Tg antibodies in ~1 mm diameter spots. Non-competitive immunoassays were performed using mixture of
125
I labeled monoclonal anti-TSH and anti-Tg antibodies. Phosphorimager was used to quantify the bound radioactivity. TSH and Tg were detected with detection limit of 0.07 µIU/ml and 0.13 ng/ml respectively, which is lower than the clinically required cut-off level. The assay showed: acceptable intra-assay precision within 20 % and recovery in the range of 76–111.2 %. MI compared well with the established immunoradiometric assay (IRMA) with r = 0.98,
p
< 0.01 (n = 41). No cross-reactivity was seen between the immobilized antibodies. Although two hormones are addressed in this report, MI using PC TEM and isotopic/non-isotopic tracers has the potential for highly automated multiplexed analysis.</abstract><cop>New Delhi</cop><pub>Springer India</pub><pmid>28428694</pmid><doi>10.1007/s12291-016-0589-2</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Antibodies Biochemistry Biomedical and Life Sciences Chemistry/Food Science Hormones Immunoassay Life Sciences Microbiology Original Original Article Pathology Patient compliance Polyclonal antibodies Prognosis Thyroid cancer Thyroid gland Thyrotropin Viral antibodies |
| title | A Microarray Immunoassay for Serum Thyrotropin and Thyroglobulin Using Antibodies Immobilized on Track-Etched Membranes |
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