Quantitative immunohistochemical assay with novel digital immunostaining for comparisons of PD-L1 antibodies
One obstacle in diagnostic pathology is the harmonization of one drug-one diagnostic tests for programmed death ligand-1 (PD-L1). There are many challenges in accurate comparisons of diagnostic tests, such as differences in the titer of each antibody, detection system and dynamic range of visualizat...
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| Veröffentlicht in: | Molecular and clinical oncology 2019-03, Vol.10 (3), p.391-396 |
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| creator | Fujisawa, Takuo Tsuta, Koji Yanagimoto, Hiroaki Yagi, Masao Suzuki, Kensuke Nishikawa, Kenji Takahashi, Masaru Okada, Hisatake Nakano, Yasushi Iwai, Hiroshi |
| description | One obstacle in diagnostic pathology is the harmonization of one drug-one diagnostic tests for programmed death ligand-1 (PD-L1). There are many challenges in accurate comparisons of diagnostic tests, such as differences in the titer of each antibody, detection system and dynamic range of visualization. Our previously developed digital immunostaining technique is highly sensitive and quantitative with the ability to quantify particles that bind in a one-to-one fashion with antibody in each cell. Determining the differences in the titer of each antibody with digital immunostaining may be beneficial for future harmonized analysis. To demonstrate the accuracy of digital immunostaining, the present study compared the number of particles with ELISA and nCounter data from five cell lines. NCI-H460 exhib-ited the highest level of PD-L1 protein, followed by A549, PC-3, NCI-H1299, and NCI-H446 cells. In addition, the PD-L1 mRNA values determined by nCounter corresponded with the order of the protein levels determined by ELISA. The present study revealed that digital immunostaining for PD-L1 was highly associated with ELISA and nCounter data. Among the four antibodies tested, the titer of all but SP142 coincided with ELISA and nCounter data. These results indicated that our digital immunostaining technique may be beneficial for future harmonized analysis. |
| doi_str_mv | 10.3892/mco.2019.1801 |
| format | Article |
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There are many challenges in accurate comparisons of diagnostic tests, such as differences in the titer of each antibody, detection system and dynamic range of visualization. Our previously developed digital immunostaining technique is highly sensitive and quantitative with the ability to quantify particles that bind in a one-to-one fashion with antibody in each cell. Determining the differences in the titer of each antibody with digital immunostaining may be beneficial for future harmonized analysis. To demonstrate the accuracy of digital immunostaining, the present study compared the number of particles with ELISA and nCounter data from five cell lines. NCI-H460 exhib-ited the highest level of PD-L1 protein, followed by A549, PC-3, NCI-H1299, and NCI-H446 cells. In addition, the PD-L1 mRNA values determined by nCounter corresponded with the order of the protein levels determined by ELISA. The present study revealed that digital immunostaining for PD-L1 was highly associated with ELISA and nCounter data. Among the four antibodies tested, the titer of all but SP142 coincided with ELISA and nCounter data. These results indicated that our digital immunostaining technique may be beneficial for future harmonized analysis.</description><identifier>ISSN: 2049-9450</identifier><identifier>EISSN: 2049-9469</identifier><identifier>DOI: 10.3892/mco.2019.1801</identifier><identifier>PMID: 30847180</identifier><language>eng</language><publisher>England: Spandidos Publications</publisher><subject>Antibodies ; Apoptosis ; Death ; Detection equipment ; Diagnostic tests ; Enzyme-linked immunosorbent assay ; Gene expression ; Immunoassay ; Immunoglobulins ; Immunohistochemistry ; Messenger RNA ; Methods ; Microscopy ; Oncology ; Protein expression ; Proteins ; RNA ; Visualization</subject><ispartof>Molecular and clinical oncology, 2019-03, Vol.10 (3), p.391-396</ispartof><rights>COPYRIGHT 2019 Spandidos Publications</rights><rights>Copyright Spandidos Publications UK Ltd. 2019</rights><rights>Copyright: © Fujisawa et al. 2019</rights><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c482t-15e2f74064d7132fb65136179b769b97240b487d5b967e3cfe3a9d9a43fbbd7a3</citedby><cites>FETCH-LOGICAL-c482t-15e2f74064d7132fb65136179b769b97240b487d5b967e3cfe3a9d9a43fbbd7a3</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/PMC6388504/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6388504/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30847180$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Fujisawa, Takuo</creatorcontrib><creatorcontrib>Tsuta, Koji</creatorcontrib><creatorcontrib>Yanagimoto, Hiroaki</creatorcontrib><creatorcontrib>Yagi, Masao</creatorcontrib><creatorcontrib>Suzuki, Kensuke</creatorcontrib><creatorcontrib>Nishikawa, Kenji</creatorcontrib><creatorcontrib>Takahashi, Masaru</creatorcontrib><creatorcontrib>Okada, Hisatake</creatorcontrib><creatorcontrib>Nakano, Yasushi</creatorcontrib><creatorcontrib>Iwai, Hiroshi</creatorcontrib><title>Quantitative immunohistochemical assay with novel digital immunostaining for comparisons of PD-L1 antibodies</title><title>Molecular and clinical oncology</title><addtitle>Mol Clin Oncol</addtitle><description>One obstacle in diagnostic pathology is the harmonization of one drug-one diagnostic tests for programmed death ligand-1 (PD-L1). There are many challenges in accurate comparisons of diagnostic tests, such as differences in the titer of each antibody, detection system and dynamic range of visualization. Our previously developed digital immunostaining technique is highly sensitive and quantitative with the ability to quantify particles that bind in a one-to-one fashion with antibody in each cell. Determining the differences in the titer of each antibody with digital immunostaining may be beneficial for future harmonized analysis. To demonstrate the accuracy of digital immunostaining, the present study compared the number of particles with ELISA and nCounter data from five cell lines. NCI-H460 exhib-ited the highest level of PD-L1 protein, followed by A549, PC-3, NCI-H1299, and NCI-H446 cells. In addition, the PD-L1 mRNA values determined by nCounter corresponded with the order of the protein levels determined by ELISA. The present study revealed that digital immunostaining for PD-L1 was highly associated with ELISA and nCounter data. Among the four antibodies tested, the titer of all but SP142 coincided with ELISA and nCounter data. These results indicated that our digital immunostaining technique may be beneficial for future harmonized analysis.</description><subject>Antibodies</subject><subject>Apoptosis</subject><subject>Death</subject><subject>Detection equipment</subject><subject>Diagnostic tests</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Gene expression</subject><subject>Immunoassay</subject><subject>Immunoglobulins</subject><subject>Immunohistochemistry</subject><subject>Messenger RNA</subject><subject>Methods</subject><subject>Microscopy</subject><subject>Oncology</subject><subject>Protein expression</subject><subject>Proteins</subject><subject>RNA</subject><subject>Visualization</subject><issn>2049-9450</issn><issn>2049-9469</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><recordid>eNptks1rHCEYxofS0oQ0x16L0Esvs_VrRr0UQvqRwEISaM-iju4aZnSrzpb893XIdtuU6MEX_T2PvPo0zVsEV4QL_HEycYUhEivEIXrRnGJIRStoL14e6w6eNOc538M6BIO4E6-bEwI5ZVVz2ox3swrFF1X83gI_TXOIW59LNFs7eaNGoHJWD-CXL1sQ4t6OYPCbyo8HOBflgw8b4GICJk47lXyOIYPowO3ndo3A4q_j4G1-07xyasz2_LCeNT--fvl-edWub75dX16sW0M5Li3qLHaMwp4ODBHsdN8h0iMmNOuFFgxTqClnQ6dFzywxzhIlBqEocVoPTJGz5tOj727Wkx2MDSWpUe6Sn1R6kFF5-fQk-K3cxL3sCecdpNXgw8EgxZ-zzUVOPhs7jirYOGeJERcd5oSIir7_D72Pcwq1vYXiiAgE0V9qo0YrfXCx3msWU3nRMd5xglBfqdUzVJ3D8hcxWOfr_hNB-ygwKeacrDv2iKBcEiJrQuSSELkkpPLv_n2YI_0nD-Q3coW3XQ</recordid><startdate>20190301</startdate><enddate>20190301</enddate><creator>Fujisawa, Takuo</creator><creator>Tsuta, Koji</creator><creator>Yanagimoto, Hiroaki</creator><creator>Yagi, Masao</creator><creator>Suzuki, Kensuke</creator><creator>Nishikawa, Kenji</creator><creator>Takahashi, Masaru</creator><creator>Okada, Hisatake</creator><creator>Nakano, Yasushi</creator><creator>Iwai, Hiroshi</creator><general>Spandidos Publications</general><general>Spandidos Publications UK Ltd</general><general>D.A. 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There are many challenges in accurate comparisons of diagnostic tests, such as differences in the titer of each antibody, detection system and dynamic range of visualization. Our previously developed digital immunostaining technique is highly sensitive and quantitative with the ability to quantify particles that bind in a one-to-one fashion with antibody in each cell. Determining the differences in the titer of each antibody with digital immunostaining may be beneficial for future harmonized analysis. To demonstrate the accuracy of digital immunostaining, the present study compared the number of particles with ELISA and nCounter data from five cell lines. NCI-H460 exhib-ited the highest level of PD-L1 protein, followed by A549, PC-3, NCI-H1299, and NCI-H446 cells. In addition, the PD-L1 mRNA values determined by nCounter corresponded with the order of the protein levels determined by ELISA. The present study revealed that digital immunostaining for PD-L1 was highly associated with ELISA and nCounter data. Among the four antibodies tested, the titer of all but SP142 coincided with ELISA and nCounter data. These results indicated that our digital immunostaining technique may be beneficial for future harmonized analysis.</abstract><cop>England</cop><pub>Spandidos Publications</pub><pmid>30847180</pmid><doi>10.3892/mco.2019.1801</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Antibodies Apoptosis Death Detection equipment Diagnostic tests Enzyme-linked immunosorbent assay Gene expression Immunoassay Immunoglobulins Immunohistochemistry Messenger RNA Methods Microscopy Oncology Protein expression Proteins RNA Visualization |
| title | Quantitative immunohistochemical assay with novel digital immunostaining for comparisons of PD-L1 antibodies |
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