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Methods. 2012 Sep;58(1):62-8. doi: 10.1016/j.ymeth.2012.07.009. Epub 2012 Jul 20.

Anti-idiotypic monobodies for immune response profiling.

Author information

1
Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA. mark_sullivan@urmc.rochester.edu

Abstract

A major goal in the study of autoimmune disease is the identification of biomarkers of disease to allow early diagnosis and initiation of treatment. The production of autoantibodies is the key feature of most autoimmune disease, so much effort has focused on characterizing the antigens reactive with these antibodies. However, even for the most well understood autoimmune diseases like rheumatoid arthritis and systemic lupus erythematosus, identification of antigens that detect autoantibodies in all patients have yet to be discovered. We describe a novel strategy for deriving mimotopes to disease-specific serum antibodies by selecting anti-idiotypic monobodies from a large molecular diversity library. Monobodies are derived by partial randomization of two surface exposed loops of a fibronectin domain scaffold in a phage display vector. The phage library is selected for binding to serum antibodies using a subtractive panning strategy. We evaluated this strategy by selecting the monobody library on a pool of serum immunoglobulin derived from a group of rheumatoid arthritis patients and evaluated selected clones for multi-patient reactivity and specificity for rheumatoid arthritis. The use of the fibronectin scaffold to derive stable, easy to produce molecular probes for diagnosis of autoimmune disease could be of significant value in improving diagnostic assays for virtually any disease that exhibits a characteristic immune response.

PMID:
22819856
PMCID:
PMC3491107
DOI:
10.1016/j.ymeth.2012.07.009
[Indexed for MEDLINE]
Free PMC Article
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