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. 2022 Oct 7;14(19):4200.
doi: 10.3390/polym14194200.

Molecularly Imprinted Ligand-Free Nanogels for Recognizing Bee Venom-Originated Phospholipase A2 Enzyme

Anamaria Zaharia  1 , Ana-Mihaela Gavrila  1 , Iuliana Caras  2 , Bogdan Trica  1 , Anita-Laura Chiriac  1 , Catalina Ioana Gifu  1 , Iulia Elena Neblea  1 , Elena-Bianca Stoica  1 , Sorin Viorel Dolana  1 , Tanta-Verona Iordache  1
Affiliations

Molecularly Imprinted Ligand-Free Nanogels for Recognizing Bee Venom-Originated Phospholipase A2 Enzyme

Anamaria Zaharia et al. Polymers (Basel). .

Abstract

In this study, ligand-free nanogels (LFNGs) as potential antivenom mimics were developed with the aim of preventing hypersensitivity and other side effects following massive bee attacks. For this purpose, poly (ethylene glycol) diacrylate was chosen as a main synthetic biocompatible matrix to prepare the experimental LFNGs. The overall concept uses inverse mini-emulsion polymerization as the main route to deliver nanogel caps with complementary cavities for phospholipase A2 (PLA2) from bee venom, created artificially with the use of molecular imprinting (MI) technologies. The morphology and the hydrodynamic features of the nanogels were confirmed by transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis. The following rebinding experiments evidenced the specificity of molecularly imprinted LFNG for PLA2, with rebinding capacities up to 8-fold higher compared to the reference non-imprinted nanogel, while the in vitro binding assays of PLA2 from commercial bee venom indicated that such synthetic nanogels are able to recognize and retain the targeted PLA2 enzyme. The results were finally collaborated with in vitro cell-viability experiments and resulted in a strong belief that such LFNG may actually be used for future therapies against bee envenomation.

Keywords: bee envenomation; bee venom phospholipase A2; ligand-free nanogels; molecularly imprinted polymers; synthetic antivenom.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Scheme 1
Scheme 1
Preparation of molecularly imprinted ligand-free nanogels (MIP-LFNGs) for recognizing and retaining bee venom-originated PLA2.
Figure 1
Figure 1
Fourier transform infrared (FT-IR) spectra of NIP-LFNG; PLA2 enzyme (PLA2); and MIP-LFNG (W or T) and MIP-LFNG (W, ext or T, ext) before and after PLA2 extraction, respectively.
Figure 2
Figure 2
(a)Thermal analysis diagram (TGA) and (b) derivative (DTG) of NIP-LFNG; PLA2 enzyme (PLA2); and MIP-LFNG (T) and MIP-LFNG (T, ext) before and after PLA2 extraction, respectively.
Figure 3
Figure 3
TEM micrograph of the NIP-LFNG ((a) 1 μm, (b) 500 nm for the final emulsion; (c) 500 nm after the washing process), imprinted MIP-LFNG (W) ((d) 1 μm and (e) 500 nm for the emulsion; (f) 500 nm after PLA2 extraction) and MIP-LFNG (T) ((g) 1 μm and (h) 500 nm for the emulsion; (i) 500 nm after PLA2 extraction).
Figure 4
Figure 4
Rebinding capacity, Q, and imprinting factor, F, of LFNGs exposed to PLA2 solution after 15 and 30 min and evaluated at (a) 450 nm and (b) 490 nm.
Figure 5
Figure 5
The drop in PLA2 activity after LFNG exposure to venom at 15 and 30 min and evaluated at (a) 450 nm and (b) 490 nm.
Figure 6
Figure 6
Cell viability of L929 cell line after 24 h exposure to various dilutions of LFNGs compared to a reference (100% viability).
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