Simple and rapid enzyme‐linked immunosorbent assay for the detection of hemoglobin C[α2β26(A3)Glu → Lys] in cord blood using a monoclonal antibody
We have generated a murine hybridoma that secretes a monoclonal antibody (mAb) that is highly specific for hemoglobin C (HbC) [α2ß26(A3)Glu→Lys] and shows no cross reactivity with HbA2, HbF2, HbS, HbE, or Hb O‐Arab. Using this antibody, we developed a simple and rapid enzyme linked immunosorbent ass...
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| Veröffentlicht in: | American journal of hematology 1990-03, Vol.33 (3), p.198-204 |
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| container_title | American journal of hematology |
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| creator | Shyamala, M. Kiefer, Charles R. Moscoso, Hugo Garver, Fred A. |
| description | We have generated a murine hybridoma that secretes a monoclonal antibody (mAb) that is highly specific for hemoglobin C (HbC) [α2ß26(A3)Glu→Lys] and shows no cross reactivity with HbA2, HbF2, HbS, HbE, or Hb O‐Arab. Using this antibody, we developed a simple and rapid enzyme linked immunosorbent assay (ELISA) technique for the detection of HbC in both adult and cord blood. The assay can be carried out using either whole blood samples or hemolysates. With as little as 10 μl/well of whole blood or 5 μg Hb/well of hemolysates, and, with dilutions of the antibody up to 10−5 we were able to detect HbC unequivocally in cord blood samples. The ELISA procedure could detect HbC in proportions as low as 0.01%. This simple diagnostic test represents a technological advance in Hb identification and can easily be used for mass screening (96 samples in less than 45 min) to detect HbC. Furthermore, this assay, when employed in conjunction with an mAb specific for β6GLU of HbA, allows the discrimination between HbC homozygotes, heterozygotes, and normals. |
| doi_str_mv | 10.1002/ajh.2830330308 |
| format | Article |
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Using this antibody, we developed a simple and rapid enzyme linked immunosorbent assay (ELISA) technique for the detection of HbC in both adult and cord blood. The assay can be carried out using either whole blood samples or hemolysates. With as little as 10 μl/well of whole blood or 5 μg Hb/well of hemolysates, and, with dilutions of the antibody up to 10−5 we were able to detect HbC unequivocally in cord blood samples. The ELISA procedure could detect HbC in proportions as low as 0.01%. This simple diagnostic test represents a technological advance in Hb identification and can easily be used for mass screening (96 samples in less than 45 min) to detect HbC. Furthermore, this assay, when employed in conjunction with an mAb specific for β6GLU of HbA, allows the discrimination between HbC homozygotes, heterozygotes, and normals.</description><identifier>ISSN: 0361-8609</identifier><identifier>EISSN: 1096-8652</identifier><identifier>DOI: 10.1002/ajh.2830330308</identifier><identifier>PMID: 2405650</identifier><identifier>CODEN: AJHEDD</identifier><language>eng</language><publisher>New York: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Anemias. Hemoglobinopathies ; Animals ; Antibodies, Monoclonal ; Biological and medical sciences ; Diseases of red blood cells ; ELISA ; Enzyme-Linked Immunosorbent Assay - methods ; Fetal Blood ; Hematologic and hematopoietic diseases ; Hemoglobin C - analysis ; Hemoglobin C - immunology ; hemoglobin C disease ; Hybridomas - immunology ; infant ; mass screening ; Medical sciences ; Mice ; Mice, Inbred BALB C ; newborn diseases ; Phenotype ; Sensitivity and Specificity</subject><ispartof>American journal of hematology, 1990-03, Vol.33 (3), p.198-204</ispartof><rights>Copyright © 1990 Wiley‐Liss, Inc., A Wiley Company</rights><rights>1990 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c2848-64f0d491c9af91b291c6eaaa7a055458d235e1a42972b7af205d97e4d8a1ca833</citedby><cites>FETCH-LOGICAL-c2848-64f0d491c9af91b291c6eaaa7a055458d235e1a42972b7af205d97e4d8a1ca833</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fajh.2830330308$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fajh.2830330308$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=6863174$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2405650$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Shyamala, M.</creatorcontrib><creatorcontrib>Kiefer, Charles R.</creatorcontrib><creatorcontrib>Moscoso, Hugo</creatorcontrib><creatorcontrib>Garver, Fred A.</creatorcontrib><title>Simple and rapid enzyme‐linked immunosorbent assay for the detection of hemoglobin C[α2β26(A3)Glu → Lys] in cord blood using a monoclonal antibody</title><title>American journal of hematology</title><addtitle>Am J Hematol</addtitle><description>We have generated a murine hybridoma that secretes a monoclonal antibody (mAb) that is highly specific for hemoglobin C (HbC) [α2ß26(A3)Glu→Lys] and shows no cross reactivity with HbA2, HbF2, HbS, HbE, or Hb O‐Arab. Using this antibody, we developed a simple and rapid enzyme linked immunosorbent assay (ELISA) technique for the detection of HbC in both adult and cord blood. The assay can be carried out using either whole blood samples or hemolysates. With as little as 10 μl/well of whole blood or 5 μg Hb/well of hemolysates, and, with dilutions of the antibody up to 10−5 we were able to detect HbC unequivocally in cord blood samples. The ELISA procedure could detect HbC in proportions as low as 0.01%. This simple diagnostic test represents a technological advance in Hb identification and can easily be used for mass screening (96 samples in less than 45 min) to detect HbC. Furthermore, this assay, when employed in conjunction with an mAb specific for β6GLU of HbA, allows the discrimination between HbC homozygotes, heterozygotes, and normals.</description><subject>Anemias. Hemoglobinopathies</subject><subject>Animals</subject><subject>Antibodies, Monoclonal</subject><subject>Biological and medical sciences</subject><subject>Diseases of red blood cells</subject><subject>ELISA</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Fetal Blood</subject><subject>Hematologic and hematopoietic diseases</subject><subject>Hemoglobin C - analysis</subject><subject>Hemoglobin C - immunology</subject><subject>hemoglobin C disease</subject><subject>Hybridomas - immunology</subject><subject>infant</subject><subject>mass screening</subject><subject>Medical sciences</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>newborn diseases</subject><subject>Phenotype</subject><subject>Sensitivity and Specificity</subject><issn>0361-8609</issn><issn>1096-8652</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc9rFDEUx4Moda1evQk5iNjDrklmJpMcl0XbyoIH9SQyvEky3dRMsiYzyHjy5Ln01H-j_iH9I_qXGNmlehMevAffz_vB-yL0lJIFJYS9gvPNgomCFDmIuIdmlEg-F7xi99GMFJzmmsiH6FFK54RQWgpygA5YSSpekRm6em_7rTMYvMYRtlZj479Pvbn9ceGs_2I0tn0_-pBCbI0fMKQEE-5CxMPGYG0GowYbPA4d3pg-nLnQWo9Xn26u2c0vxl8ui6NjN-Lbn5d4PaXPOIsqRI1bF4LGY7L-DAPugw_KBQ8uHzLYNujpMXrQgUvmyT4foo9vXn9YnczX745PV8v1XDFRijkvO6JLSZWETtKW5YobAKiBVFVZCc2KylAomaxZW0PHSKVlbUotgCoQRXGIXuzmbmP4Opo0NL1NyjgH3oQxNbXkRBApMrjYgSqGlKLpmm20PcSpoaT5Y0WTrWj-WpEbnu0nj21v9B2-_33Wn-91SApcF8Erm-4wLnhB6zJjcod9s85M_1naLN-e_HPCbxN4piI</recordid><startdate>199003</startdate><enddate>199003</enddate><creator>Shyamala, M.</creator><creator>Kiefer, Charles R.</creator><creator>Moscoso, Hugo</creator><creator>Garver, Fred A.</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley-Liss</general><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>7X8</scope></search><sort><creationdate>199003</creationdate><title>Simple and rapid enzyme‐linked immunosorbent assay for the detection of hemoglobin C[α2β26(A3)Glu → Lys] in cord blood using a monoclonal antibody</title><author>Shyamala, M. ; Kiefer, Charles R. ; Moscoso, Hugo ; Garver, Fred A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c2848-64f0d491c9af91b291c6eaaa7a055458d235e1a42972b7af205d97e4d8a1ca833</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>Anemias. Hemoglobinopathies</topic><topic>Animals</topic><topic>Antibodies, Monoclonal</topic><topic>Biological and medical sciences</topic><topic>Diseases of red blood cells</topic><topic>ELISA</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Fetal Blood</topic><topic>Hematologic and hematopoietic diseases</topic><topic>Hemoglobin C - analysis</topic><topic>Hemoglobin C - immunology</topic><topic>hemoglobin C disease</topic><topic>Hybridomas - immunology</topic><topic>infant</topic><topic>mass screening</topic><topic>Medical sciences</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>newborn diseases</topic><topic>Phenotype</topic><topic>Sensitivity and Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Shyamala, M.</creatorcontrib><creatorcontrib>Kiefer, Charles R.</creatorcontrib><creatorcontrib>Moscoso, Hugo</creatorcontrib><creatorcontrib>Garver, Fred A.</creatorcontrib><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>MEDLINE - Academic</collection><jtitle>American journal of hematology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Shyamala, M.</au><au>Kiefer, Charles R.</au><au>Moscoso, Hugo</au><au>Garver, Fred A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Simple and rapid enzyme‐linked immunosorbent assay for the detection of hemoglobin C[α2β26(A3)Glu → Lys] in cord blood using a monoclonal antibody</atitle><jtitle>American journal of hematology</jtitle><addtitle>Am J Hematol</addtitle><date>1990-03</date><risdate>1990</risdate><volume>33</volume><issue>3</issue><spage>198</spage><epage>204</epage><pages>198-204</pages><issn>0361-8609</issn><eissn>1096-8652</eissn><coden>AJHEDD</coden><abstract>We have generated a murine hybridoma that secretes a monoclonal antibody (mAb) that is highly specific for hemoglobin C (HbC) [α2ß26(A3)Glu→Lys] and shows no cross reactivity with HbA2, HbF2, HbS, HbE, or Hb O‐Arab. Using this antibody, we developed a simple and rapid enzyme linked immunosorbent assay (ELISA) technique for the detection of HbC in both adult and cord blood. The assay can be carried out using either whole blood samples or hemolysates. With as little as 10 μl/well of whole blood or 5 μg Hb/well of hemolysates, and, with dilutions of the antibody up to 10−5 we were able to detect HbC unequivocally in cord blood samples. The ELISA procedure could detect HbC in proportions as low as 0.01%. This simple diagnostic test represents a technological advance in Hb identification and can easily be used for mass screening (96 samples in less than 45 min) to detect HbC. Furthermore, this assay, when employed in conjunction with an mAb specific for β6GLU of HbA, allows the discrimination between HbC homozygotes, heterozygotes, and normals.</abstract><cop>New York</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>2405650</pmid><doi>10.1002/ajh.2830330308</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0361-8609 |
| ispartof | American journal of hematology, 1990-03, Vol.33 (3), p.198-204 |
| issn | 0361-8609 1096-8652 |
| language | eng |
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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | Anemias. Hemoglobinopathies Animals Antibodies, Monoclonal Biological and medical sciences Diseases of red blood cells ELISA Enzyme-Linked Immunosorbent Assay - methods Fetal Blood Hematologic and hematopoietic diseases Hemoglobin C - analysis Hemoglobin C - immunology hemoglobin C disease Hybridomas - immunology infant mass screening Medical sciences Mice Mice, Inbred BALB C newborn diseases Phenotype Sensitivity and Specificity |
| title | Simple and rapid enzyme‐linked immunosorbent assay for the detection of hemoglobin C[α2β26(A3)Glu → Lys] in cord blood using a monoclonal antibody |
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