Naturally Occurring Anti-albumin Antibodies Are Responsible for False Positivity in Diagnosis of Autoimmune Premature Ovarian Failure
Autoimmunity is a well-established causative factor of premature ovarian failure (POF), and evidence for the same has been well reported in the literature. Detection of specific autoantibodies remains the most practical clinical research marker of any autoimmune disease. Variation in efficiency and...
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Veröffentlicht in: | The journal of histochemistry and cytochemistry 2006-04, Vol.54 (4), p.397-405 |
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creator | Pires, Eusebio S Parte, Priyanka P Meherji, Perveen K Khan, Shagufta A Khole, Vrinda V |
description | Autoimmunity is a well-established causative factor of premature ovarian failure (POF), and evidence for the same has been well reported in the literature. Detection of specific autoantibodies remains the most practical clinical research marker of any autoimmune disease. Variation in efficiency and specificity in the detection of ovarian autoantibodies has been reported. However, the frequency of false positivity and a solution to overcome this has not yet been reported. Herein, we report autoantibody to albumin as the likely responsible agent for false positivity. Our data indicate that presence of naturally existing autoalbumin antibodies in the circulation of normal women is responsible for the false signal seen in SDS-PAGE Western blot analysis and in immunohistochemistry (IHC). Having shown the presence of anti-albumin antibody in normal women as well as in the sera of POF patients, we have developed a novel blocking agent to overcome this problem. A high titer polyclonal antibody against human serum albumin was generated. This antibody showed immunoreactivity to albumin obtained from various sources. Preincubation of Western blots and IHC sections with this antibody drastically reduced background signals. The advantage of using this blocking was evident by identification of specific anti-ovarian antibodies in a group of POF patients. This blocking procedure made it possible to obtain a clear indication of the ovarian antibody status in women presenting with autoimmune POF. |
doi_str_mv | 10.1369/jhc.5A6778.2005 |
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Detection of specific autoantibodies remains the most practical clinical research marker of any autoimmune disease. Variation in efficiency and specificity in the detection of ovarian autoantibodies has been reported. However, the frequency of false positivity and a solution to overcome this has not yet been reported. Herein, we report autoantibody to albumin as the likely responsible agent for false positivity. Our data indicate that presence of naturally existing autoalbumin antibodies in the circulation of normal women is responsible for the false signal seen in SDS-PAGE Western blot analysis and in immunohistochemistry (IHC). Having shown the presence of anti-albumin antibody in normal women as well as in the sera of POF patients, we have developed a novel blocking agent to overcome this problem. A high titer polyclonal antibody against human serum albumin was generated. This antibody showed immunoreactivity to albumin obtained from various sources. Preincubation of Western blots and IHC sections with this antibody drastically reduced background signals. The advantage of using this blocking was evident by identification of specific anti-ovarian antibodies in a group of POF patients. This blocking procedure made it possible to obtain a clear indication of the ovarian antibody status in women presenting with autoimmune POF.</description><identifier>ISSN: 0022-1554</identifier><identifier>EISSN: 1551-5044</identifier><identifier>DOI: 10.1369/jhc.5A6778.2005</identifier><identifier>PMID: 16286665</identifier><language>eng</language><publisher>Los Angeles, CA: Histochemical Soc</publisher><subject>Adult ; Animals ; Autoantibodies - blood ; Autoimmune Diseases - diagnosis ; Autoimmune Diseases - immunology ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; False Positive Reactions ; Female ; Humans ; Immunohistochemistry ; Male ; Ovary - immunology ; Primary Ovarian Insufficiency - diagnosis ; Primary Ovarian Insufficiency - immunology ; Rabbits ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Serum Albumin - immunology</subject><ispartof>The journal of histochemistry and cytochemistry, 2006-04, Vol.54 (4), p.397-405</ispartof><rights>2006 Authors</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c403t-76f973f054f6f72b72ab83b1bd110062cca16416162f93ad4c1cb12f20e73f03</citedby><cites>FETCH-LOGICAL-c403t-76f973f054f6f72b72ab83b1bd110062cca16416162f93ad4c1cb12f20e73f03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://journals.sagepub.com/doi/pdf/10.1369/jhc.5A6778.2005$$EPDF$$P50$$Gsage$$H</linktopdf><linktohtml>$$Uhttps://journals.sagepub.com/doi/10.1369/jhc.5A6778.2005$$EHTML$$P50$$Gsage$$H</linktohtml><link.rule.ids>314,776,780,21799,27903,27904,43600,43601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16286665$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pires, Eusebio S</creatorcontrib><creatorcontrib>Parte, Priyanka P</creatorcontrib><creatorcontrib>Meherji, Perveen K</creatorcontrib><creatorcontrib>Khan, Shagufta A</creatorcontrib><creatorcontrib>Khole, Vrinda V</creatorcontrib><title>Naturally Occurring Anti-albumin Antibodies Are Responsible for False Positivity in Diagnosis of Autoimmune Premature Ovarian Failure</title><title>The journal of histochemistry and cytochemistry</title><addtitle>J Histochem Cytochem</addtitle><description>Autoimmunity is a well-established causative factor of premature ovarian failure (POF), and evidence for the same has been well reported in the literature. Detection of specific autoantibodies remains the most practical clinical research marker of any autoimmune disease. Variation in efficiency and specificity in the detection of ovarian autoantibodies has been reported. However, the frequency of false positivity and a solution to overcome this has not yet been reported. Herein, we report autoantibody to albumin as the likely responsible agent for false positivity. Our data indicate that presence of naturally existing autoalbumin antibodies in the circulation of normal women is responsible for the false signal seen in SDS-PAGE Western blot analysis and in immunohistochemistry (IHC). Having shown the presence of anti-albumin antibody in normal women as well as in the sera of POF patients, we have developed a novel blocking agent to overcome this problem. A high titer polyclonal antibody against human serum albumin was generated. This antibody showed immunoreactivity to albumin obtained from various sources. Preincubation of Western blots and IHC sections with this antibody drastically reduced background signals. The advantage of using this blocking was evident by identification of specific anti-ovarian antibodies in a group of POF patients. This blocking procedure made it possible to obtain a clear indication of the ovarian antibody status in women presenting with autoimmune POF.</description><subject>Adult</subject><subject>Animals</subject><subject>Autoantibodies - blood</subject><subject>Autoimmune Diseases - diagnosis</subject><subject>Autoimmune Diseases - immunology</subject><subject>Blotting, Western</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>False Positive Reactions</subject><subject>Female</subject><subject>Humans</subject><subject>Immunohistochemistry</subject><subject>Male</subject><subject>Ovary - immunology</subject><subject>Primary Ovarian Insufficiency - diagnosis</subject><subject>Primary Ovarian Insufficiency - immunology</subject><subject>Rabbits</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>Reproducibility of Results</subject><subject>Serum Albumin - immunology</subject><issn>0022-1554</issn><issn>1551-5044</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc1uGyEURlGUKnF-1tlVbNJVxgEGGHs5Spu2UlRXVfYIMNhYM-DCEMsP0Pcuk7HUXVfoovN96B4AuMNojmu-fNxt9Zy1vGkWc4IQOwMzzBiuGKL0HMwQIqQqF_QSXKW0QwhTyhYX4BJzsuCcsxn480MOOcquO8KV1jlG5zew9YOrZKdy7_z7oMLamQTbaOAvk_bBJ6c6A22I8Fl2ycCfIbnBvbnhCEvks5MbX24SDBa2eQiu77MvVDT9-JyBqzcZnfQl7boy34APduy5PZ3X4PX5y-vTt-pl9fX7U_tSaYrqoWq4XTa1RYxabhuiGiLVolZYrTFGiBOtJeYU87KeXdZyTTXWChNLkBlj9TX4NNXuY_idTRpE75I2XSe9CTmJ4rGIqUkBHydQx5BSNFbso-tlPAqMxCheFPFiEi9G8SXx8VSdVW_W__iT6QI8TECSGyN2IUdfNv1P3_2Eb91me3DRiNSXXyrtWBwOB0YFFXWR8RdWGJve</recordid><startdate>20060401</startdate><enddate>20060401</enddate><creator>Pires, Eusebio S</creator><creator>Parte, Priyanka P</creator><creator>Meherji, Perveen K</creator><creator>Khan, Shagufta A</creator><creator>Khole, Vrinda V</creator><general>Histochemical Soc</general><general>SAGE Publications</general><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>20060401</creationdate><title>Naturally Occurring Anti-albumin Antibodies Are Responsible for False Positivity in Diagnosis of Autoimmune Premature Ovarian Failure</title><author>Pires, Eusebio S ; Parte, Priyanka P ; Meherji, Perveen K ; Khan, Shagufta A ; Khole, Vrinda V</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c403t-76f973f054f6f72b72ab83b1bd110062cca16416162f93ad4c1cb12f20e73f03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Adult</topic><topic>Animals</topic><topic>Autoantibodies - blood</topic><topic>Autoimmune Diseases - diagnosis</topic><topic>Autoimmune Diseases - immunology</topic><topic>Blotting, Western</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>False Positive Reactions</topic><topic>Female</topic><topic>Humans</topic><topic>Immunohistochemistry</topic><topic>Male</topic><topic>Ovary - immunology</topic><topic>Primary Ovarian Insufficiency - diagnosis</topic><topic>Primary Ovarian Insufficiency - immunology</topic><topic>Rabbits</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>Reproducibility of Results</topic><topic>Serum Albumin - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pires, Eusebio S</creatorcontrib><creatorcontrib>Parte, Priyanka P</creatorcontrib><creatorcontrib>Meherji, Perveen K</creatorcontrib><creatorcontrib>Khan, Shagufta A</creatorcontrib><creatorcontrib>Khole, Vrinda V</creatorcontrib><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>The journal of histochemistry and cytochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pires, Eusebio S</au><au>Parte, Priyanka P</au><au>Meherji, Perveen K</au><au>Khan, Shagufta A</au><au>Khole, Vrinda V</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Naturally Occurring Anti-albumin Antibodies Are Responsible for False Positivity in Diagnosis of Autoimmune Premature Ovarian Failure</atitle><jtitle>The journal of histochemistry and cytochemistry</jtitle><addtitle>J Histochem Cytochem</addtitle><date>2006-04-01</date><risdate>2006</risdate><volume>54</volume><issue>4</issue><spage>397</spage><epage>405</epage><pages>397-405</pages><issn>0022-1554</issn><eissn>1551-5044</eissn><abstract>Autoimmunity is a well-established causative factor of premature ovarian failure (POF), and evidence for the same has been well reported in the literature. Detection of specific autoantibodies remains the most practical clinical research marker of any autoimmune disease. Variation in efficiency and specificity in the detection of ovarian autoantibodies has been reported. However, the frequency of false positivity and a solution to overcome this has not yet been reported. Herein, we report autoantibody to albumin as the likely responsible agent for false positivity. Our data indicate that presence of naturally existing autoalbumin antibodies in the circulation of normal women is responsible for the false signal seen in SDS-PAGE Western blot analysis and in immunohistochemistry (IHC). Having shown the presence of anti-albumin antibody in normal women as well as in the sera of POF patients, we have developed a novel blocking agent to overcome this problem. A high titer polyclonal antibody against human serum albumin was generated. This antibody showed immunoreactivity to albumin obtained from various sources. Preincubation of Western blots and IHC sections with this antibody drastically reduced background signals. The advantage of using this blocking was evident by identification of specific anti-ovarian antibodies in a group of POF patients. This blocking procedure made it possible to obtain a clear indication of the ovarian antibody status in women presenting with autoimmune POF.</abstract><cop>Los Angeles, CA</cop><pub>Histochemical Soc</pub><pmid>16286665</pmid><doi>10.1369/jhc.5A6778.2005</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Adult Animals Autoantibodies - blood Autoimmune Diseases - diagnosis Autoimmune Diseases - immunology Blotting, Western Electrophoresis, Polyacrylamide Gel False Positive Reactions Female Humans Immunohistochemistry Male Ovary - immunology Primary Ovarian Insufficiency - diagnosis Primary Ovarian Insufficiency - immunology Rabbits Rats Rats, Sprague-Dawley Reproducibility of Results Serum Albumin - immunology |
title | Naturally Occurring Anti-albumin Antibodies Are Responsible for False Positivity in Diagnosis of Autoimmune Premature Ovarian Failure |
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