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KMID : 0545120070170060993
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Journal of Microbiology and Biotechnology
2007 Volume.17 No. 6 p.993 ~ p.1001
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Optimized Serological Isolation of Lung-Cancer-associated Antigens from a Yeast Surface-expressed cDNA Library
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Kim Min-Soo
Choi Hye-Young
Choi Yong-Soo
Kim Jhin-Gook
Kim Young-Sun
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Abstract
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The technique of serological analysis of antigens by recombinant cDNA expression library (SEREX) uses autologous patient sera as a screening probe to isolate tumorassociated antigens for various tumor types. Isolation of tumor-associated antigens that are specifically reactive with patient sera, but not with normal sera, is important to avoid false-positive and autoimmunogenic antigens for the cancer immunotherapy. Here, we describe a selection methodology to isolate patient sera-specific antigens from a yeast surfaceexpressed cDNA library constructed from 15 patient lung tissues with non-small cell lung cancer (NSCLC). Several rounds of positive selection using patient sera alone as a screening probe isolated clones exhibiting comparable reactivity with both patient and normal sera. However, the combination of negative selection with allogeneic normal sera to remove antigens reactive with normal sera and subsequent positive selection with patient sera efficiently enriched patient sera-specific antigens. Using the selection methodology described here, we isolated 3 known and 5 unknown proteins, which have not been isolated previously, but and potentially associated with NSCLC.
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KEYWORD
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SEREX, yeast surface display, cDNA library, tumor-associated antigens, non-small cell lung cancer
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