Screening and characterization of human scFv antibodies against nasopharyngeal carcinoma

2009 
Objective To screen the anti-nasopharyngeal carcinoma scFv from a human anti-nasopharyngeal carcinoma single-chain phage antibody library, and identify its characteristics. Methods The single-chain phage antibody library was subjected to three rounds of positive and negative cell panning and enrichment, and then it was selected by ELISA. The binding specificity of phage antibodies with naso-pharyngeal carcinoma cells was confirmed by immunohistochemistry. Results After panning, enrichment and testing by ELISA, 3 phage an-tibody clones reacting with CNE2 more strongly than HUVEC and NP69 were picked out from 4212 clones. One clone, HNSAO33, was fur-ther analyzed after DNA sequencing. The results of immunohistochemistry with cultured cells were similar to those of ELISA. HNSAO33 spe-cifically reacted to nasopharyngeal carcinoma cells in most human nasopharyngeal carcinoma tissue sections except a few human normal naso-pharyngeal tissue sections. The distinction of positive rates was of a great statistical significance. Conclusion ELISA and immunohisto-chemistry results confirmed HNSAO33 specifically bind with nasopharyngeal carcinoma cells. The seFv fragment against nasopharyngeal carci-noma may be further developed and applied in clinical diagnosis and therapy of nasopharyngeal carcinoma. Key words: Bacteriophages/IM;  Antibodies, monoclonal/BI;  Nasopharyngeal neoplasms/IM
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