Density Matching Multi-wavelength Analytical Ultracentrifugation to Measure Drug Loading of Lipid Nanoparticle Formulations

ACS Nano ◽  
2021 ◽  
Author(s):  
Amy Henrickson ◽  
Jayesh A. Kulkarni ◽  
Josh Zaifman ◽  
Gary E. Gorbet ◽  
Pieter R. Cullis ◽  
...  
Keyword(s):  
Analytical Ultracentrifugation ◽  
Drug Loading ◽  
Lipid Nanoparticle ◽  
Multi Wavelength

Formulation Development of Tamoxifen Loaded Lipid Nanoparticle by Taguchi (L12 (2 11)) Orthogonal Array Design

2020 ◽  
Vol 16 ◽  
Author(s):  
Poovi Ganesan ◽  
N Damodharan
Keyword(s):  
Orthogonal Array ◽  
Drug Loading ◽  
Optimization Technique ◽  
Pharmaceutical Products ◽  
Water Soluble ◽  
Nanostructured Lipid Carrier ◽  
Orthogonal Array Design ◽  
Formulation Development ◽  
Array Design ◽  
Lipid Nanoparticle

Background: A better understanding of the biopharmaceutical and physicochemical properties of drugs and the pharmaco-technical factors would be of great help for developing pharmaceutical products. But, it is extremely difficult to study the effect of each variable and interaction among them through the conventional approach Objective: To screen the most influential factors affecting the particle size (PS) of lipid nanoparticle (LNPs) (solid lipid nanoparticle (SLN) and nanostructured lipid carrier (NLC)) for poorly water-soluble BCS class-II drug like tamoxifen (TMX) to improve its oral bioavailability and to reduce its toxicity to tolerable limits using Taguchi (L12 (2 11)) orthogonal array design by applying computer optimization technique. Results: The size of all LNPs formulations prepared as per the experimental design varied between 172 nm and 3880 μm, polydispersity index between 0.033 and 1.00, encapsulation efficiency between 70.8% and 75.7%, and drug loading between 5.84% and 9.68%. The study showed spherical and non-spherical as well as aggregated and non-aggregated LNPs. Besides, it showed no interaction and amorphous form of the drug in LNPs formulation. The Blank NLCs exhibited no cytotoxicity on MCF-7 cells as compared to TMX solution, SLNs (F5) and NLCs (F12) suggests that the cause of cell death is primarily from the effect of TMX present in NLCs. Conclusions: The screening study clearly showed the importance of different individual factors significant effect for the LNPs formulation development and its overall performance in an in-vitro study with minimum experimentation thus saving considerable time, efforts, and resources for further in-depth study.

scholarly journals Formulation of Solid Lipid Nanoparticles of Cilnidipine for the Treatment of Hypertension

2019 ◽  
Vol 9 (3) ◽  
pp. 212-221 ◽  
Author(s):  
Aparna Bhalerao ◽  
Pankaj Prakash Chaudhari
Keyword(s):  
Particle Size ◽  
Solid Lipid Nanoparticles ◽  
Encapsulation Efficiency ◽  
Drug Loading ◽  
Lipid Nanoparticles ◽  
Water Soluble ◽  
Solid Lipid Nanoparticle ◽  
Solid Lipid ◽  
Lipid Nanoparticle

Cilinidipine is a fourth generation N and L-type calcium channel antagonists used alone or in combination with another drug to treat hypertension. Cilnidipine is poorly water -soluble, BCS class II drug with 6 to 30 percent oral bioavailability due to first pass metabolism. So to protect the drug from degradation and improve its dissolution, solid lipid nanoparticles were prepared. Glyceryl monostearate was selected as lipid while span 20: tween 20 were selected as surfactant blends. The formulations were evaluated for various parameters, as percent transmittance, drug content, percent encapsulation efficiency; percent drug loading, In vitro drug release and particle size. Optimized formulation was lyophilized using lactose as a cryo-protectant. The lyophilized formulation was evaluated for micromeritic properties, particle size and in vitro dissolution. It was further evaluated for DSC, XRD, and SEM. Percent encapsulation efficiency and percent drug loading of optimized formulation (F3) were 78.66percent and 9.44percent respectively. The particle size of F3 formulation without drug was 204 nm and with the drug was 214 nm. The particle size of the reconstituted SLN was 219 nm. In DSC study, no obvious peaks for cilnidipine were found in the SLN of cilnidipine indicated that the cilnidipine must be present in a molecularly dissolved state in SLN. In X-ray diffractometry absence of peaks representing crystals of cilnidipine in SLN indicated that the drug was in an amorphous or disordered crystalline phase in the lipid matrix. Thus, solid lipid nanoparticle formulation is a promising way to enhance the dissolution rate of cilnidipine. Keywords: Cilnidipine, Solid Lipid Nanoparticle, Hypertension

scholarly journals Multi-Wavelength Analytical Ultracentrifugation of Biopolymer Mixtures and Interactions

2021 ◽  
Author(s):  
Borries Demeler ◽  
Amy Henrickson ◽  
Gary Gorbet ◽  
Alexey Savelyev ◽  
Minji Kim ◽  
...  
Keyword(s):  
Small Molecules ◽  
Analytical Ultracentrifugation ◽  
Optical Systems ◽  
Hydrodynamic Properties ◽  
Advantages And Disadvantages ◽  
Design Considerations ◽  
Free Dna ◽  
Wide Range ◽  
Analysis Strategies ◽  
Multi Wavelength

Abstract Multi-wavelength analytical ultracentrifugation (MW-AUC) is a recent development made possible by new analytical ultracentrifuge optical systems. MW-AUC is suitable for a wide range of applications and biopolymer systems and is poised to become an essential tool to characterize macromolecular interactions. It adds an orthogonal spectral dimension to the traditional hydrodynamic characterization by exploiting unique chromophores in analyte mixtures that may or may not interact. Here we illustrate the utility of MW-AUC for representative classes of challenging biopolymer systems, including interactions between mixtures of different sized proteins with small molecules, mixtures of loaded and empty viral AAV capsids contaminated with free DNA, and mixtures of different proteins, where some have identical hydrodynamic properties, all of which are difficult to resolve with traditional AUC methods. We explain the improvement in resolution and information content obtained by this technique compared to traditional single- or dual-wavelength approaches. We discuss experimental design considerations and limitations of the method, and address the advantages and disadvantages of the two MW optical systems available today, and the differences in data analysis strategies between the two systems.

Size-Dependent Drug Loading, Gene Complexation, Cell Uptake, and Transfection of a Novel Dendron-Lipid Nanoparticle for Drug/Gene Co-delivery

2021 ◽  
Author(s):  
Ashita Nair ◽  
Jiyoon Bu ◽  
Jason Bugno ◽  
Piper A. Rawding ◽  
Luke J. Kubiatowicz ◽  
...  
Keyword(s):  
Drug Loading ◽  
Cell Uptake ◽  
Size Dependent ◽  
Lipid Nanoparticle

Multi-wavelength analytical ultracentrifugation as a tool to characterise protein–DNA interactions in solution

2020 ◽  
Vol 49 (8) ◽  
pp. 819-827 ◽  
Author(s):  
Christopher R. Horne ◽  
Amy Henrickson ◽  
Borries Demeler ◽  
Renwick C. J. Dobson
Keyword(s):  
Analytical Ultracentrifugation ◽  
Dna Interactions ◽  
Protein Dna Interactions ◽  
Multi Wavelength

scholarly journals Measuring molecular interactions in solution using multi-wavelength analytical ultracentrifugation: combining spectral analysis with hydrodynamics

2019 ◽  
Vol 41 (2) ◽  
pp. 14-18 ◽  
Author(s):  
Borries Demeler
Keyword(s):  
Analytical Ultracentrifugation ◽  
Heterogeneous Systems ◽  
Colloidal Properties ◽  
Resolution Capability ◽  
Technological Advances ◽  
Data Density ◽  
Hydrodynamic Separation ◽  
Multi Wavelength ◽  
Spectral Absorbance ◽  
Indispensable Tool

In 1926, the Swedish scientist Theodor Svedberg was awarded the Nobel Prize in Chemistry for his work on a disperse system, and for studying the colloidal properties of proteins. This work was, to a large extent, made possible by his invention of a revolutionary tool, the analytical ultracentrifuge. These days, technological advances in hardware and computing have transformed the field of analytical ultracentrifugation (AUC) by enabling entirely new classes of experiments and modes of measurement unimaginable by Svedberg, making AUC once again an indispensable tool for modern biomedical research. In this article these advances and their impact on studies of interacting molecules will be discussed, with particular emphasis on a new method termed multi-wavelength analytical ultracentrifugation (MWL-AUC). Novel detectors allow us to add a second dimension to the separation of disperse and heterogeneous systems: in addition to the traditional hydrodynamic separation of colloidal mixtures, it is now possible to identify the sedimenting molecules by their spectral absorbance properties. The potential for this advance is significant for the study of a large range of systems. A further advance has occurred in data management and computational capabilities, opening doors to improved analysis methods, as well as direct networking with the instrument, facilitating data acquisition and data handling, and significant increases in data density from faster detectors with higher resolution capability.

scholarly journals Consensus model of a cyanobacterial light-dependent protochlorophyllide oxidoreductase in its pigment-free apo-form and photoactive ternary complex

2019 ◽  
Vol 2 (1) ◽  
Author(s):  
Judith Schneidewind ◽  
Frank Krause ◽  
Marco Bocola ◽  
Andreas Maximilian Stadler ◽  
Mehdi D. Davari ◽  
...  
Keyword(s):  
Ternary Complex ◽  
Analytical Ultracentrifugation ◽  
Structural Information ◽  
Md Simulations ◽  
Consensus Model ◽  
Photosynthetic Organisms ◽  
X Ray Scattering ◽  
Multi Wavelength ◽  
Modelling Studies ◽  
Chlorophyll Precursor

Abstract Photosynthetic organisms employ two different enzymes for the reduction of the C17 = C18 double bond of protochlorophyllide (Pchlide), yielding the chlorophyll precursor chlorophyllide. First, a nitrogenase-like, light-independent (dark-operative) Pchlide oxidoreductase and secondly, a light-dependent Pchlide oxidoreductase (LPOR). For the latter enzyme, despite decades of research, no structural information is available. Here, we use protein structure modelling, molecular dynamics (MD) simulations combined with multi-wavelength analytical ultracentrifugation (MWA-AUC) and small angle X-ray scattering (SAXS) experiments to derive a consensus model of the LPOR apoprotein and the substrate/cofactor/LPOR ternary complex. MWA-AUC and SAXS experiments independently demonstrate that the apoprotein is monomeric, while ternary complex formation induces dimerization. SAXS-guided modelling studies provide a full-length model of the apoprotein and suggest a tentative mode of dimerization for the LPOR ternary complex, supported by published cross-link constraints. Our study provides a first impression of the LPOR structural organization.

scholarly journals Multi‐wavelength Analytical Ultracentrifugation of Human Serum Albumin complexed with Porphyrin

2018 ◽  
Vol 32 (S1) ◽  
Author(s):  
Courtney Nicole Johnson ◽  
Gary E. Gorbet ◽  
Heidi Ramsower ◽  
Julio Uriquidi ◽  
Lorenzo Brancaleon ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Analytical Ultracentrifugation ◽  
Multi Wavelength

scholarly journals Multi-Wavelength Analytical Ultracentrifugation of Biopolymer Mixtures and Interactions

2021 ◽  
Author(s):  
Amy Henrickson ◽  
Gary E Gorbet ◽  
Alexey Savelyev ◽  
Minji Kim ◽  
Sarah K Schultz ◽  
...  
Keyword(s):  
Small Molecules ◽  
Analytical Ultracentrifugation ◽  
Optical Systems ◽  
Hydrodynamic Properties ◽  
Advantages And Disadvantages ◽  
Design Considerations ◽  
Free Dna ◽  
Wide Range ◽  
Analysis Strategies ◽  
Multi Wavelength

Multi-wavelength analytical ultracentrifugation (MW-AUC) is a recent development made possible by new analytical ultracentrifuge optical systems. MW-AUC is suitable for a wide range of applications and biopolymer systems and is poised to become an essential tool to characterize macromolecular interactions. It adds an orthogonal spectral dimension to the traditional hydrodynamic characterization by exploiting unique chromophores in analyte mixtures that may or may not interact. Here we illustrate the utility of MW-AUC for representative classes of challenging biopolymer systems, including interactions between mixtures of different sized proteins with small molecules, mixtures of loaded and empty viral AAV capsids contaminated with free DNA, and mixtures of different proteins, where some have identical hydrodynamic properties, all of which are difficult to resolve with traditional AUC methods. We explain the improvement in resolution and information content obtained by this technique compared to traditional single- or dual-wavelength approaches. We discuss experimental design considerations and limitations of the method, and address the advantages and disadvantages of the two MW optical systems available today, and the differences in data analysis strategies between the two systems.

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