doi:10.1369/jhc.5A6778.2005
Volume 54 (4): 397-405, 2006 Copyright ©The Histochemical Society, Inc. Naturally Occurring Anti-albumin Antibodies Are Responsible for False Positivity in Diagnosis of Autoimmune Premature Ovarian Failure
Department of Gamete Immunobiology (ESP,PPP,SAK,VVK) and Department of Endocrinology and Infertility (PKM), National Institute for Research in Reproductive Health, Mumbai, India Correspondence to: Dr. Vrinda V. Khole, PhD, Deputy Director and Divisional Head, Department of Gamete Immuno Biology, National Institute for Research in Reproductive Health (ICMR), J M Street, Parel, Mumbai 400 012, India. E-mail: kholevv{at}icmr.org.in 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. (J Histochem Cytochem 54:397405, 2006)
Key Words: autoimmune premature ovarian failure false positives blocking agents autoantibodies anti-albumin antibodies
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