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Functional stability of a nucleic acid-hydrolyzing single chain antibody with anti-viral activity in various biochemical and biophysical environments

Authors
Lee, Joungmin; Kim, Minjae; Park, Hyunjoon; Seo, Youngsil; Lee, Yeonjin; Kwon, Myung-Hee
Department
Department of Microbiology
Abstract
3D8 single-chain Fv (scFv), a recombinant catalytic anti-nucleic acid antibody with nucleic acid-hydrolyzing activities, is known to have potential anti-viral activity against broad spectrum of viruses. Here, we investigate the functional of 3D8 scFv by assessing DNA-hydrolyzing and –binding activity in various physical and chemical conditions. 3D8 scFv exposed at 50°C and pH 7.2 for 6 hrs sustained the DNA-hydrolyzing activity, while DNA-binding activity was reduced by 50%. Optimum pH for both activities of 3D8 scFv ranged from 7 to 11. When one-week exposed to various pH, both activities of 3D8 scFv were fully recovered in the samples exposed at pH 3, 4, 5, and 10. Activities of 3D8 scFv were not affected in reducing condition built by 1 mM DTT, by freezing-thawing up to 30 cycles, and lyophilization. 3D8 scFv were functionally stable for 3-week and 1-week storage in pH 7.2 at 23°C and 37°C, respectively. Furthermore, we determined the correlation between the units of DNA-hydrolyzing activity and anti-viral effect of 3D8 scFv at cell level. Our study would provide the basis for the formulation of 3D8 scFv to develop it as the anti-viral agent.
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