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Annals of Clinical & Laboratory Science 35:121-130 (2005)
© 2005 Association of Clinical Scientists

Methodologic Factors Affect the Measurement of Anti-basal Ganglia Antibodies

Christopher A. Rippel1, John J. Hong1, Dustin Y. Yoon1, Phillip N. Williams1 and Harvey S. Singer1,2
1 Departments of Neurology and 2 Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland

Address correspondence to Harvey S. Singer M.D., Division of Pediatric Neurology, Johns Hopkins Hospital, Jefferson Street Building 124, 600 N. Wolfe Street, Baltimore, MD 21287-1000, USA; tel 410 955 7212; fax 410 614 2297; e-mail:hsinger{at}jhmi.edu.

An autoimmune etiology has been proposed for a variety of movement disorders, making the detection of autoantibodies a high investigative priority. Recognizing the existence of different methodologic approaches to identify these antibodies, we sought to investigate the effects of tissue preparation, antibody selection, and Western immunoblot detection methods on outcome. ELISA and immunoblotting studies were performed in healthy controls evaluating non-pathogenic autoantibodies. Our results indicate that enhanced data can be obtained by using fresh, rather than frozen, postmortem tissue homogenates for Western immunoblots and enzyme-linked immunosorbent assays and support the use of electrochemiluminescent detection for Western immunoblots. Molecular localization is significantly affected by the selected standard. Removal of lipids from homogenates does not affect anti-basal ganglia antibody (ABGA) results. Methodological variables should be taken into consideration when performing and interpreting neuroimmunological assays using sera or isolated IgG.

Keywords: autoantibodies, ELISA, immunological methods, Western immunoblots




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