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At least 19 records

Excipient screening by lyophilization provides insights into spray drying formulations for nanoparticle vaccines

Nanoparticles have shown great promise as delivery platforms in the development of tunable and safe vaccines. Nanolipoprotein particles (NLPs), also known as nanodiscs, are discoidal nanoparticles composed of a lipid bilayer stabilized at their periphery by apolipoproteins. Under the right conditions, the NLP self-assembly process is highly customizable in terms of lipids and apolipoprotein constituents, allowing for tunable physical and chemical characteristics. This flexibility allows a wide range of vaccine antigens and adjuvants to be incorporated onto the NLP platform for tailored vaccine design. The stability of NLPs during long term storage is a very important factor in developing a vaccine delivery platform suitable for widespread global use. When stored in a solution for extended periods of time, NLPs dissociate into their corresponding lipids and protein constituents, leading to particle degradation. Proper stabilization of NLPs can often be achieved by lyophilization (i.e. freeze-drying), a method widely used for various applications including pharmaceuticals. This process, however, can be damaging to particles without the presence of lyoprotectants or excipients that help maintain particle stability during lyophilization. Another method used to stabilize vaccines and pharmaceuticals is spray drying, a process that converts liquid formulations into dry powders through controlled heating and airflow. While spray drying is rapid, scalable, and cost-effective, lyophilization is typically a gentler process that better retains biomolecule structure and function. Both processes use excipients for particle stabilization, so lyophilization can be used as a surrogate to test stability of NLPs, to down-select formulations that may withstand the harsher conditions of spray drying. To screen different formulations, NLPs were synthesized and purified to homogeneity by size exclusion chromatography (SEC) and samples were prepared with a wide range of lyoprotectants and/or excipients. To assess the protective effects of excipients on NLPs upon spray drying, both pre- and post-lyophilized samples were analyzed by SEC. To assess the protective effects upon heating (encountered during the spray drying process), NLP samples were incubated at elevated temperatures prior to SEC analysis. The lyoprotectants and excipients evaluated in this study had different efficiencies in protecting NLPs during lyophilization and heating tests. Trehalose, for example, exhibits stabilization on NLPs both upon lyophilization and heating whereas leucine accelerated NLP dissociation. Although some lyoprotectants are effective by themselves, different combinations can decrease the stabilization of NLPs. Shelf-stable vaccines that do not require cold-chain storage are essential for global accessibility and our findings provide fundamental insight into how to advance NLP-based vaccines for these applications.

Serrano, Litzay J [Lawrence Livermore National Lab↗

Lyophilization of ASFV vaccine candidate ASFV-G-ΔI177L offers long term stability

Abstract For over a century African swine fever (ASF) has been causing outbreaks leading to devastating losses for the swine industry. The current pandemic of ASF has shown no signs of stopping and continues to spread causing outbreaks in additional countries. Currently control relies mostly on culling infected farms, and strict biosecurity procedures. Recently a vaccine, ASFV-G-ΔI177L was approved for use in Vietnam. In this study we evaluate the long-term stability of lyophilized ASFV-G-ΔI177L. Understanding the stability of different formulations of vaccines is information necessary for deployment of vaccines to ASF outbreak areas, particularly those that do not have a reliable well established cold chain to ensure conservation of vaccine quality. In this report, we determined that ASFV-G-ΔI177L, when lyophilized under specific conditions, is stable for up to one year at 4 °C, with similar vaccine titers after storage. Next-generation sequencing analysis also determined that lyophilization and long-term storage under these conditions had no effect on the genome of ASFV as the genome remained genetically identical to the original non-lyophilized form.

Science & Technology - Other Topics↗

Lyophilization for Water Recovery From Solid Waste

This abstract describes the development of a solid waste treatment system designed for a near term human exploration mission. The technology being developed is an energy- efficient lyophilization technique that recovers water from spacecraft solid waste. In the lyophilization process water in an aqueous waste is frozen and then sublimed, resulting in the separation of the waste into a dried solid material and liquid water. This technology is ideally suited to applications where water recovery rates approaching 100% are desirable but production of CO, is not. Water contained within solid wastes accounts for approximately 3% of the total water balance. If 100% closure of the water loop is desired the water contained within this waste would need to be recovered. To facilitate operation in microgravity thermoelectric heat pumps have be used in place of traditional fluid cycle heat pumps. A mathematical model of a thermoelectric lyophilizer has been developed and used to generate energy use and processing rate parameters. The results of laboratory investigations and discussions with ALS program management have been used to iteratively arrive at a prototype design. This design address operational limitations which were identified in the laboratory studies and handling and health concerns raised by ALS program management. The current prototype design is capable of integration into the ISS Waste Collection System.

Flynn, Michael↗

Lyophilization -Solid Waste Treatment

This paper discusses the development of a solid waste treatment system that has been designed for a Mars transit exploration mission. The technology described is an energy-efficient lyophilization technique that is designed to recover water from spacecraft solid wastes. Candidate wastes include feces, concentrated brines from water processors, and other solid wastes that contain free water. The system is designed to operate as a stand-alone process or to be integrated into the International Space Station Waste Collection System. In the lyophilization process, water in an aqueous waste is frozen and then sublimed, separating the waste into a dried solid material and liquid water. The sublimed water is then condensed in a solid ice phase and then melted to generate a liquid product. In the subject system the waste solids are contained within a 0.2 micron bio-guard bag and after drying are removed from the system and stored in a secondary container. This technology is ideally suited to applications such as the Mars Reference Mission, where water recovery rates approaching 100% are desirable but production of CO2 is not. The system is designed to minimize power consumption through the use of thermoelectric heat pumps. The results of preliminary testing of a prototype system and testing of the final configuration are provided. A mathematical model of the system is also described.

Litwiller, Eric↗

PCR amplification of 16S rDNA from lyophilized cell cultures facilitates studies in molecular systematics

The sequence of the major portion of a Bacillus cycloheptanicus strain SCH(T) 16S rRNA gene is reported. This sequence suggests that B. cycloheptanicus is genetically quite distinct from traditional Bacillus strains (e.g., B. subtilis) and may be properly regarded as belonging to a different genus. The sequence was determined from DNA that was produced by direct amplification of ribosomal DNA from a lyophilized cell pellet with straightforward polymerase chain reaction (PCR) procedures. By obviating the need to revive cell cultures from the lyophile pellet, this approach facilitates rapid 16S rDNA sequencing and thereby advances studies in molecular systematics.

Non-NASA Center↗

Design and Testing of a Lyophilizer for Water Recovery from Solid Waste

Mixed liquid/solid wastes, including feces, water processor effluents, and food waste, can be lyophilized (freeze-dried) to recover the water they contain and stabilize the solids remain. Previous research has demonstrated the potential benefits of using thermoelectric heat pumps to build a lyophilizer for processing waste in microgravity. These results were used to build a working prototype suitable for ground-based human testing. This paper describes the prototype design and presents the results of functional and performance tests. Equivalent system mass parameters are calculated, and practical issues such as sanitary waste handling in microgravity are addressed.

Litwiller, Eric↗

Lyophilization for Water Recovery III, System Design

Mixed liquid/solid wastes, including feces, water processor effluents, and food waste, can be lyophilized (freeze-dried) to recover the water they contain and stabilize the solids that remain. Our previous research has demonstrated the potential benefits of using thermoelectric heat pumps to build a lyophilizer for processing waste in microgravity. These results were used to build a working prototype suitable for ground- based human testing. This paper describes the prototype design and presents results of functional and performance tests.

Litwiller, Eric↗

Modeling of Heat and Mass Transfer in a TEC-Driven Lyophilizer

Dewatering of wet waste during space exploration missions is important for crew safety as it stabilizes the waste. It may also be used to recover water and serve as a preconditioning step for waste compaction. A thermoelectric cooler (TEC)-driven lyophilizer is under development at NASA Ames Research Center for this purpose. It has three major components: (i) an evaporator section where water vapor sublimes from the frozen waste, (ii) a condenser section where this water vapor deposits as ice, and (iii) a TEC section which serves as a heat pump to transfer heat from the condenser to the evaporator. This paper analyses the heat and mass transfer processes in the lyophilizer in an effort to understand the ice formation behavior in the condenser. The analysis is supported by experimental observations of ice formation patterns in two different condenser units.

Yuan, Zeng-Guang↗

Lyophilized spore dispenser

A lyophilized spore dispenser is provided which produces a finely divided, monoparticulate cloud of bacterial spores. The spores are contained within a tightly sealed chamber, and a turbulator orifice connected to an air supply source provides a jet of air which stirs up the spores and causes the spores to be suspended in eddy currents within the chamber. This air jet also produces a positive pressure within the chamber which forces the spores out of an injection orifice.

Jessup, A. D.↗

Combustion water purification techniques influence on OBT analysing using liquid scintillation counting method

In order to determine organically bound tritium (OBT) from environmental samples, these must be converted into water, measurable by liquid scintillation counting (LSC). For this purpose we conducted some experiments to determine OBT level of a grass sample collected from an uncontaminated area. The studied grass sample was combusted in a Parr bomb. However usual interfering phenomena were identified: color or chemical quench, chemiluminescence, overlap over tritium spectrum because of other radionuclides presence as impurities ({sup 14}C from organically compounds, {sup 36}Cl as chloride and free chlorine, {sup 40}K as potassium cations) and emulsion separation. So the purification of the combustion water before scintillation counting appeared to be essential. 5 purification methods were tested: distillation with chemical treatment (Na{sub 2}O{sub 2} and KMnO{sub 4}), lyophilization, chemical treatment (Na{sub 2}O{sub 2} and KMnO{sub 4}) followed by lyophilization, azeotropic distillation with toluene and treatment with a volcanic tuff followed by lyophilization. After the purification step each sample was measured and the OBT measured concentration, together with physico-chemical analysis of the water analyzed, revealed that the most efficient method applied for purification of the combustion water was the method using chemical treatment followed by lyophilization.

54 ENVIRONMENTAL SCIENCES↗

Effect of Storage Conditions on the Integrity of Human Umbilical Cord Mesenchymal Stromal Cell-Derived Microvesicles

We studied the effect of storage conditions on the safety of microvesicles produced by human multipotent umbilical cord mesenchymal stromal cells into the conditioned medium. It was found that microvesicles can be stored without serious degradation for up to 1 week at 4°С, but were almost completely destroyed during freezing and thawing cycles irrespective of the storage temperatures (-20°С, -70°С, or -196°С). Similar results were obtained for lyophilized medium conditioned by human multipotent umbilical cord mesenchymal stromal cells. Addition of a cryoprotectant (5-10% DMSO) followed by freezing and/or lyophilization preserved microvesicles at a nearly initial level. These findings indicate that during storage, microvesicles, being membrane structures, behave similar to living cells and require appropriate conditions for prolonged storage.

60 APPLIED LIFE SCIENCES↗

Wetting properties of molecularly rough surfaces

We employ molecular dynamics simulations to study the wettability of nanoscale rough surfaces in systems governed by Lennard-Jones (LJ) interactions. We consider both smooth and molecularly rough planar surfaces. Solid substrates are modeled as a static collection of LJ particles arranged in a face-centered cubic lattice with the (100) surface exposed to the LJ fluid. Molecularly rough solid surfaces are prepared by removing several strips of LJ atoms from the external layers of the substrate, i.e., forming parallel nanogrooves on the surface. We vary the solid-fluid interactions to investigate strongly and weakly wettable surfaces. We determine the wetting properties by measuring the equilibrium droplet profiles that are in turn used to evaluate the contact angles. Macroscopic arguments, such as those leading to Wenzel’s law, suggest that surface roughness always amplifies the wetting properties of a lyophilic surface. However, our results indicate the opposite effect from roughness for microscopically corrugated surfaces, i.e., surface roughness deteriorates the substrate wettability. Adding the roughness to a strongly wettable surface shrinks the surface area wet with the liquid, and it either increases or only marginally affects the contact angle, depending on the degree of liquid adsorption into the nanogrooves. For a weakly wettable surface, the roughness changes the surface character from lyophilic to lyophobic due to a weakening of the solid-fluid interactions by the presence of the nanogrooves and the weaker adsorption of the liquid into the nanogrooves.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗