Version 2 2024-01-30, 23:10Version 2 2024-01-30, 23:10
Version 1 2024-01-29, 11:29Version 1 2024-01-29, 11:29
journal contribution
posted on 2024-01-30, 23:10authored byCecilia Contardi, Davide Rubes, Massimo Serra, Rossella Dorati, Marco Dattilo, Liliia Mavliutova, Maddalena Patrini, Raffaella Guglielmann, Börje Sellergren, Ersilia De Lorenzi
In
this work, an innovative and accurate affinity capillary
electrophoresis
(ACE) method was set up to monitor the complexation of aqueous MIP
nanogels (NGs) with model cancer-related antigens. Using α2,6′-
and α2,3′-sialyllactose as oversimplified cancer biomarker-mimicking
templates, NGs were synthesized and characterized in terms of size,
polydispersity, and overall charge. A stability study was also carried
out in order to select the best storage conditions and to ensure product
quality. After optimization of capillary electrophoresis conditions,
injection of MIP NGs resulted in a single, sharp, and efficient peak.
The mobility shift approach was applied to quantitatively estimate
binding affinity, in this case resulting in an association constant
of K ≈ 106 M–1. The optimized polymers further displayed a pronounced discrimination
between the two sialylated sugars. The newly developed ACE protocol
has the potential to become a very effective method for nonconstrained
affinity screening of NG in solution, especially during the NG development
phase and/or for a final accurate quantitation of the observed binding.