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1.
Biotechnol Bioeng ; 118(8): 3251-3262, 2021 08.
Article in English | MEDLINE | ID: mdl-34129733

ABSTRACT

Due to the high variation in viral surface properties, a platform method for virus purification is still lacking. A potential alternative to the high-cost conventional methods is aqueous two-phase systems (ATPSs). However, optimizing virus purification in ATPS requires a large experimental design space, and the optimized systems are generally found to operate at high ATPS component concentrations. The high concentrations capitalize on hydrophobic and electrostatic interactions to obtain high viral particle yields. This study investigated using osmolytes as driving force enhancers to reduce the high concentration of ATPS components while maintaining high yields. The partitioning behavior of porcine parvovirus (PPV), a nonenveloped mammalian virus, and human immunodeficiency virus-like particle (HIV-VLP), a yeast-expressed enveloped VLP, were studied in a polyethylene glycol (PEG) 12 kDa-citrate system. The partitioning of the virus modalities was enhanced by osmoprotectants glycine and betaine, while trimethylamine N-oxide was ineffective for PPV. The increased partitioning to the PEG-rich phase pertained only to viruses, resulting in high virus purification. Recoveries were 100% for infectious PPV and 92% for the HIV-VLP, with high removal of the contaminant proteins and more than 60% DNA removal when glycine was added. The osmolyte-induced ATPS demonstrated a versatile method for virus purification, irrespective of the expression system.


Subject(s)
HIV-1/isolation & purification , Parvovirus, Porcine/isolation & purification , Virion/isolation & purification , Animals , Cell Line , HIV-1/chemistry , Humans , Parvovirus, Porcine/chemistry , Swine , Virion/chemistry
2.
Plants (Basel) ; 8(5)2019 Apr 26.
Article in English | MEDLINE | ID: mdl-31027336

ABSTRACT

Alternative food supplies could maintain humanity despite sun-blocking global catastrophic risks (GCRs) that eliminate conventional agriculture. A promising alternative food is making leaf concentrate. However, the edibility of tree leaves is largely uncertain. To overcome this challenge, this study provides the methods for obtaining rapid toxics screening of common leaf concentrates. The investigation begins with a non-targeted approach using an ultra-high-resolution hybrid ion trap orbitrap mass spectrometer with electrospray ionization (ESI) coupled to an ultra-high pressure two-dimensional liquid chromatograph system on the most common North American leaf: the red maple. Identified chemicals from this non-targeted approach are then cross-referenced with the OpenFoodTox database to identify toxic chemicals. Identified toxins are then screened for formula validation and evaluated for risk as a food. The results after screening show that red maple leaf concentrate contains at least eight toxic chemicals, which upon analysis do not present substantial risks unless consumed in abundance. This indicates that red maple leaf is still a potential alternative food. The results are discussed in the context of expanding the analysis with open science and using leaf extract from other plants that are not traditionally used as foods to offset current global hunger challenges, and move to a more sustainable food system while also preparing for GCRs.

3.
Water Res ; 135: 22-30, 2018 05 15.
Article in English | MEDLINE | ID: mdl-29454238

ABSTRACT

Considering the increasing identification of trace organic contaminants in natural aquatic environments, the removal of trace organic contaminants from water or wastewater discharge is an urgent task. Ultraviolet (UV) and UV-based advanced oxidation processes (AOPs), such as UV/hydrogen peroxide (UV/H2O2), UV/free chlorine and UV/persulfate, are attractive and promising approaches for the removal of these contaminants due to the high reactivity of active radical species produced in these UV-AOPs with a wide variety of organic contaminants. However, the removal efficiency of trace contaminants is greatly affected by the presence of background dissolved organic matter (DOM). In this study, we use ultrahigh resolution mass spectrometry to evaluate the transformation of a standard Suwanee River fulvic acid DOM isolate in UV photolysis and UV-AOPs. The use of probe compounds allows for the determination of the steady-state concentrations of active radical species in each UV-AOP. The changes in the H/C and O/C elemental ratios, double bond equivalents, and the low-molecular-weight transformation product concentrations of organic acids reveal that different DOM transformation patterns are induced by each UV-AOP. By comparison with the known reactivities of each radical species with specific organic compounds, we mechanistically and systematically elucidate the molecular-level DOM transformation pathways induced by hydroxyl, chlorine, and sulfate radicals in UV-AOPs. We find that there is a distinct transformation in the aliphatic components of DOM due to HO• in UV/H2O2 and UV/free chlorine. Cl• induced transformation of olefinic species is also observed in the UV/free chlorine system. Transformation of aromatic and olefinic moieties by SO4•- are the predominant pathways in the UV/persulfate system.


Subject(s)
Hydroxyl Radical/chemistry , Sulfates/chemistry , Water Pollutants, Chemical/chemistry , Water Purification/methods , Benzopyrans/chemistry , Chlorine/chemistry , Hydrogen Peroxide/chemistry , Mass Spectrometry/methods , Molecular Weight , Organic Chemicals , Oxidation-Reduction , Photolysis , Ultraviolet Rays , Wastewater/chemistry
4.
Data Brief ; 11: 316-330, 2017 Apr.
Article in English | MEDLINE | ID: mdl-28275666

ABSTRACT

Two separate liquid chromatography (LC)-mass spectrometry (MS) methods were developed for determination and quantification of water-soluble and fat-soluble vitamins in human tear and blood serum samples. The water-soluble vitamin method was originally developed to detect vitamins B1, B2, B3 (nicotinamide), B5, B6 (pyridoxine), B7, B9 and B12 while the fat-soluble vitamin method detected vitamins A, D3, 25(OH)D3, E and K1. These methods were then validated with tear and blood serum samples. In this data in brief article, we provide details on the two LC-MS methods development, methods sensitivity, as well as precision and accuracy for determination of vitamins in human tears and blood serum. These methods were then used to determine the vitamin concentrations in infant and parent samples under a clinical study which were reported in "Determination of Water-Soluble and Fat-Soluble Vitamins in Tears and Blood Serum of Infants and Parents by Liquid Chromatography/Mass Spectrometry DOI:10.1016/j.exer.2016.12.007 [1]". This article provides more details on comparison of vitamin concentrations in the samples with the ranges reported in the literature along with the medically accepted normal ranges. The details on concentrations below the limits of detection (LOD) and limits of quantification (LOQ) are also discussed. Vitamin concentrations were also compared and cross-correlated with clinical data and nutritional information. Significant differences and strongly correlated data were reported in [1]. This article provides comprehensive details on the data with slight differences or slight correlations.

5.
Exp Eye Res ; 155: 54-63, 2017 02.
Article in English | MEDLINE | ID: mdl-28025000

ABSTRACT

Tears serve as a viable diagnostic fluid with advantages including less invasive sample to collect and less complex to prepare for analysis. Several water-soluble and fat-soluble vitamins were detected and quantified in human tears and compared with blood serum levels. Samples from 15 family pairs, each pair consisting of a four-month-old infant and one parent were analyzed; vitamin concentrations were compared between tears and blood serum for individual subjects, between infants and parents, and against self-reported dietary intakes. Water-soluble vitamins B1, B2, B3 (nicotinamide), B5, B9 and fat-soluble vitamin E (α-tocopherol) were routinely detected in tears and blood serum while fat-soluble vitamin A (retinol) was detected only in blood serum. Water-soluble vitamin concentrations measured in tears and blood serum of single subjects were comparable, while higher concentrations were measured in infants compared to their parents. Fat-soluble vitamin E concentrations were lower in tears than blood serum with no significant difference between infants and parents. Serum vitamin A concentrations were higher in parents than infants. Population trends were compiled and quantified using a cross correlation factor. Strong positive correlations were found between tear and blood serum concentrations of vitamin E from infants and parents and vitamin B3 concentrations from parents, while slight positive correlations were detected for infants B3 and parents B1 and B2 concentrations. Correlations between infants and parents were found for the concentrations of B1, B2, B3, and E in tears, and the concentrations of B2, A, and E in blood serum. Stronger vitamin concentration correlations were found between infants and parents for the breast-fed infants, while no significant difference was observed between breast-fed and bottle-fed infants. This work is the first to demonstrate simultaneous vitamin A, B, and E detection and to quantify correlations between vitamin concentrations in tears and blood serum. Our results suggest that tears are a viable biofluid to monitor nutritional health because they sufficiently mirror blood serum data and may enhance the speed of deficiency diagnoses.


Subject(s)
Chromatography, High Pressure Liquid/methods , Mass Spectrometry/methods , Parents , Serum/metabolism , Tears/metabolism , Vitamins/metabolism , Female , Humans , Infant , Male
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