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J. Biol. Chem., Vol. 279, Issue 47, 48817-48820, November 19, 2004

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Immunoglobulins in Urine of Hamsters with Scrapie^*

Ana Serban, Giuseppe Legname, Kirk Hansen¶, Nadia Kovaleva, and Stanley B. Prusiner||**

From the Institute for Neurodegenerative Diseases and Departments of Neurology, ^||Biochemistry and Biophysics, and ^¶Pharmaceutical Chemistry, University of California, San Francisco California 94143

In the prion diseases, a prolonged, asymptomatic incubation period precedes the onset of neurologic dysfunction. At present, a noninvasive test is not available for the presymptomatic diagnosis of prion disease, and thus the report of a test for prions using urine has been of great interest (Shaked, G. M., Shaked, Y., Kariv-Inbal, Z., Halimi, M., Avraham, I., and Gabizon, R. (2001) J. Biol. Chem. 276, 31479–31482). Using Western immunoblots with the anti-prion protein (PrP) 3F4 monoclonal antibody and an anti-mouse IgG secondary antibody, a protease-resistant PrP was reported in the urine of Syrian hamsters and humans with prion disease. Here we have demonstrated that this purportedly "protease-resistant PrP" band in the urine of diseased hamsters is detectable using the anti-mouse IgG secondary antibody in the absence of the 3F4 monoclonal antibody. Mass spectrometric analysis identified an immunoglobulin light chain in the band but found no PrP peptides. No similar band was found in the urine of uninfected hamsters or in brain homogenates from normal or prioninfected hamsters. Moreover, the band in the urine of infected hamsters was not detected using two chimeric human-mouse recombinant anti-PrP antibody fragments followed by an anti-human IgG secondary antibody. Our results indicate that the band detected under previously published conditions is due to the cross-reactivity of the anti-mouse IgG antibody with IgG light chains and possibly heavy chain fragments in urine, but not with PrP.

Received for publication, August 9, 2004

^* This work was supported by National Institutes of Health Grants AG02132 and AG010770 as well as by a gift from the G. Harold and Leila Y. Mathers Charitable Foundation. A. Serban, G. Legname, and S. B. Prusiner have financial interest in InPro Biotechnology. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^** To whom correspondence should be addressed: Institute for Neurodegenerative Diseases, Box 0518, University of California, San Francisco, CA 94143-0518. Tel.: 415-476-4482; Fax: 415-476-8386; E-mail: stanley{at}ind.ucsf.edu.

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