Optimizing nanoformulations for encapsulating recombinant antibodies in multiple sclerosis therapy

Introduction: Monoclonal antibodies (mAb) for multiple sclerosis (MS) therapy is one of the currently recommended treatments.Anti-VLA-4 is a successfully established bioactive for MS therapy since 2004.However, the rapid degradation of mAbs in the body is a known issue.Nanoencapsulation can protect mAb from degradation, improve permeation capacity and site-specific targeting.Polymeric nanoparticles (PNs) have gained prominence in the field of drug delivery due to their biodegradability, controlled release and greater bioavailability compared to free bioactives. Objectives:To establish an optimal condition for the recombinant mAb encapsulation, aiming to a future formulation using a new recombinant anti-VLA-4.Methodology: PNs were synthesized by double emulsion (DE) technique using of 1, 3 and 5 mg/ml of mAb.The encapsulation efficiency (EE) and loading capacity (LC) were obtained by spectrophotometry.PNs characterization was realized by Dynamic Light Scattering (DLS).After the synthesis process and drug release assay, structural and homogenity evaluations of the mAb were done by fluorescence spectroscopy and gel electrophoresis (SDS-PAGE). Results:The DLS results demonstrated PNs with approximately 204.8nm, 198.6 nm and 214.4nm, using respectively 1, 3 and 5mg/mL of mAb.The polydispersity index for all formulations were below 0.22, with Zeta potential of -7.92; -6.48 and -6.45 mV , respectively.Similar EE (over 80%) and LC (4.2) were obtained using 3 or 5mg/mL of mAb.Homogeneity and structural analysis suggest mAb characteristics were maintained, such as maximum fluorescence emission at 330 nm and electrophoretic profile, with a band of 170kDa for both conditions: prior the synthesis process and after release, since the control mAb had presented the same maximum emission and 172kDa, by SDS-PAGE.Conclusion: mAb encapsulation in polymeric matrixes showed to be efficient using 3 or 5mg/mL and with the sensitivity to be detected by spectrophotometry.Evaluation of the mAb after encapsulation process its integrity was mantained.