posted on 2016-02-22, 03:55authored byMahmoud Labib, Anna S. Zamay, Darija Muharemagic, Alexey V. Chechik, John C. Bell, Maxim V. Berezovski
DNA aptamers are promising immunoshielding agents that
could protect
oncolytic viruses (OVs) from neutralizing antibodies (nAbs) and increase
the efficiency of cancer treatment. In the present Article, we introduce
a novel technology for electrochemical differentiation of epitope-specific
aptamers (eDEA) without selecting aptamers against individual antigenic
determinants. For this purpose, we selected DNA aptamers that can
bind noncovalently to an intact oncolytic virus, vaccinia virus (VACV),
which can selectively replicate in and kill only tumor cells. The
aptamers were integrated as a recognition element into a multifunctional
electrochemical aptasensor. The developed aptasensor was used for
the linear quantification of the virus in the range of 500–3000
virus particles with a detection limit of 330 virions. Also, the aptasensor
was employed to compare the binding affinities of aptamers to VACV
and to estimate the degree of protection of VACV using the anti-L1R
neutralizing antibody in a displacement assay fashion. Three anti-VACV
aptamer clones, vac2, vac4, and vac6, showed the best immunoprotection
results and can be applied for enhanced delivery of VACV. Another
two sequences, vac5 and vac46, exhibited high affinities to VACV without
shielding it from nAb and can be further utilized in sandwich bioassays.