Nanolipoprotein Particles as a Delivery Platform for Fab Based Therapeutics
Autor: | Yvonne Franke, Kelly M. Loyet, Ryan Abraham, Martine Darwish, Kathy Barrett, May Lin, Hong Li, Christine Tam, Craig Blanchette, Inna Zilberleyb, Janice L Wong, Kyle Mortara, Brandon Leonard, Whitney Shatz |
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Rok vydání: | 2020 |
Předmět: |
Hydrodynamic radius
Lipoproteins Biomedical Engineering Pharmaceutical Science Nanoparticle Bioengineering 02 engineering and technology 01 natural sciences Maleimides chemistry.chemical_compound Immunoglobulin Fab Fragments Drug Delivery Systems In vivo Lipid bilayer Maleimide Pharmacology 010405 organic chemistry Chemistry Organic Chemistry 021001 nanoscience & nanotechnology 0104 chemical sciences Nanostructures Biophysics Particle size 0210 nano-technology Rheology Biotechnology Conjugate |
Zdroj: | Bioconjugate chemistry. 31(8) |
ISSN: | 1520-4812 |
Popis: | Nanolipoprotein particles (NLPs), a lipid bilayer-based nanoparticle platform, have recently been developed for in vivo delivery of a variety of molecules of therapeutic interest, but their potential to deliver Fabs with valencies that exceed those of current multivalent formats has not yet been evaluated. Here we describe the development, optimization, and characterization of Fab-NLP conjugates. NLPs were generated with maleimide reactive lipids for conjugation to a Fab with a C-terminal cysteine. Of note, maleimide reactive lipids were shown to conjugate to the apolipoprotein when the NLPs were assembled at pH 7.4. However, this undesirable reaction was not observed when assembled at pH 6. Site-specific Fab conjugation conditions were then optimized, and conjugation of up to 30 Fab per NLP was demonstrated. Interestingly, although conjugation of higher numbers of Fabs had a significant impact on NLP molecular weight, only a minimal impact on NLP hydrodynamic radius was observed, indicating that particle size is largely dictated by the discoidal shape of the NLP. Fab-NLP viscosity and its stability upon lyophilization were also evaluated as an assessment of the manufacturability of the Fab-NLP. Significantly higher Fab concentrations were achieved with the Fab-NLP conjugates relative to another multivalent format (Fab-PEG conjugates). Fab conjugation to the NLP was also not found to have an impact on Fab activity in both an inhibitory and agonist setting. Finally, the stability of the Fab-NLP conjugates was evaluated in 50% serum and Fab-NLPs demonstrated increased stability, with >63% of Fab-NLP remaining intact after 24 h at Fab per particle ratios of 7 or greater. Our findings suggest Fab-NLPs are a promising platform for the targeted delivery of Fabs in a multivalent format and are compatible with established manufacturing processes. |
Databáze: | OpenAIRE |
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