Affinity capillary electrophoresis to evaluate the complex formation between poliovirus and nanobodies.

It was demonstrated that nanobodies with an in vitro neutralizing activity against poliovirus
type 1 interact with native virions.Here, the use of capillary electrophoresis was investigated
as an alternative technique for the evaluation of the formation of nanobody-poliovirus
complexes, and therefore predicting the in vitro neutralizing activity of the nanobodies. The
macromolecules are preincubated offline in a specific nanobody-to-virus ratio and analyzed
by capillary electrophoresis with UV detection. At low nanobody-to-virus ratios, a clear shift
in migration time of the viral peak was observed. A broad peak was obtained, indicating
the presence of a heterogeneous population of nanobody-virion complexes, caused by the
binding of different numbers of nanobodies to the virus particle. At elevated nanobody-tovirus
ratios, a cluster of peaks appeared, showing an additional increase inmigration times.
It was shown that, at these high molar excesses, aggregates were formed. The developed
capillary electrophoresismethod can be used as a rapid, qualitative screening for the affinity
between poliovirus and nanobodies, based on a clearly visible and measurable shift in
migration time. The advantages of this technique include that there is no need for antigen
immobilization as in enzyme-linked immunosorbent assays or surface plasmon resonance
for the use of radiolabeled virus or for the performance of labor- and time-intensive plaqueforming
neutralization assays.

Jaar van publicatie:2014

Trefwoorden:Affinity, Capillary electrophoresis

CSS-citation score:1

Toegankelijkheid:Open