Nanoparticle size and production efficiency are affected by the presence of fatty acids during albumin nanoparticle fabrication

Luebbert, Christian; Clarke, Tessa M.; Pointet, Roberta; Frahm, Grant E.; Tam, Sharon; Lorbetskie, Barry; Sauvé, Simon; Johnston, Michael

doi:10.1371/journal.pone.0189814

Cited by 13 publications

(19 citation statements)

References 41 publications

(79 reference statements)

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“…However, recovery of the N -albumin shape produces reorganization of the BSA molecules, forming an external wall of nanoparticles and an internal core of crystalline capsaicin. A similar effect was observed by Luebbert et al [18] during a study on the formation of protein aggregates and increased size of dodecanoic acid-defatted BSA nanoparticles; they found that the secondary structure of BSA protein did not show changes during the fatty acid loading procedure. However, the loading efficiency (similar to EE%) and the diameter of nanoparticles were affected; they proposed a reorganization of the fatty acid in the nanoparticles as a possible mechanism that explains these changes.…”

Section: Resultssupporting

confidence: 83%

“…Choi et al [30] found that particles elaborated with Capsicum oleoresin and several different wall materials exhibited fluctuations in the diameter size of the nanoparticles. Luebbert et al [18] found that native BSA (with fatty acids) showed an increase in nanoparticle sizes with respect to fatty acid-free BSA (DF-BSA). However, the diameter of particles showed a maximal increment of diameter respect to the concentration of the dodecanoic acid (12 C in length) added during the nanostructuration process.…”

Section: Resultsmentioning

confidence: 99%

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Generation of BSA-capsaicin Nanoparticles and Their Hormesis Effect on the Rhodotorula mucilaginosa Yeast

et al. 2019

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Capsaicin is a chemical compound found in pungent chili peppers (Capsicum spp.). In biotechnology, capsaicin has been proposed as a pathogen control; however, its low solubility in water and high instability limits its uses. The aim of this work was to study the effect of high concentrations of capsaicin on the synthesis of nanoparticles and to evaluate their inhibitory effect on the growth of Rhodotorula mucilaginosa yeast. Bovine serum albumin (BSA)-capsaicin nanoparticles were formulated at 0, 16.2, 32.5, 48.7 and 65.0 µg of capsaicin per mg of BSA. Nanoparticle properties were evaluated and they were added to cultures of R. mucilaginosa to quantify their effect on cell viability. We found that increased capsaicin levels caused several changes to the physicochemical parameters, probably due to changes in the hydrophobicity sites of the albumin during the nanostructuration. The administration of nanoparticles to cultures of R. mucilaginosa produced a maximal viability with nanoparticles at 16.2 µg/mg; on the contrary, nanoparticles at 65.0 µg/mg caused maximal cell death. R. mucilaginosa cells displayed a hormesis effect in response to the nanoparticle dose concentration. The nanoparticles showed different responses during the uptake process, probably as a consequence of the nanostructural properties of capsaicin in the BSA molecules.

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Section: Resultssupporting

confidence: 83%

Section: Resultsmentioning

confidence: 99%

Generation of BSA-capsaicin Nanoparticles and Their Hormesis Effect on the Rhodotorula mucilaginosa Yeast

et al. 2019

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“…We suppose that differences in nanoparticle preparation stem from the presence of fatty acids in BSA preparations. Recently, the effect of fatty acids on the both size and yield of BSA nanoparticles was confirmed in a study by Luebbert et al (Luebbert, 2017). Fig.…”

Section: The Desolvation Process Differs When Bsa From Different Manumentioning

confidence: 71%

“…The presence of low-molecular-weight gelatin fractions led to the formation of aggregates during the synthesis of gelatin nanoparticles . Type of protein (native, recombinant) affects the size of nanoparticles, besides lot-to-lot variability also takes place (Luebbert, 2017).…”

Section: The Desolvation Process Differs When Bsa From Different Manumentioning

confidence: 99%

Measuring the Concentration of Protein Nanoparticles Synthesized by Desolvation Method: Comparison of Bradford Assay, BCA Assay, hydrolysis/UV Spectroscopy and Gravimetric Analysis

Khramtsov¹,

Калашникова²,

Бочкова³

et al. 2020

Preprint

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Research paper on sunthesis of protein nanoparticles<div><br><div><b>Abstract</b></div><div>The desolvation technique is one of the most popular methods for preparing protein nanoparticles for medicine, biotechnology, and food applications. We fabricated 11 batches of BSA nanoparticles and 2 batches of gelatin nanoparticles by desolvation method. BSA nanoparticles from 2 batches were cross-linked by heating at +70 °C for 2 h; other nanoparticles were stabilized by glutaraldehyde. We compared several analytical approaches to measuring their concentration: gravimetric analysis, bicinchoninic acid assay, Bradford assay, and alkaline hydrolysis combined with UV spectroscopy. We revealed that the cross-linking degree and method of cross-linking affect both Bradford and BCA assay. Direct measurement of protein concentration in the suspension of purified nanoparticles by dye-binding assays can lead to significant (up to 50-60%) underestimation of nanoparticle concentration. Quantification of non-desolvated protein (indirect method) is affected by the presence of small nanoparticles in supernatants and can be inaccurate when the yield of desolvation is low. The reaction of cross-linker with protein changes UV absorbance of the latter. Therefore pure protein solution is an inappropriate calibrator when applying UV spectroscopy for the determination of nanoparticle concentration. Our recommendation is to determine the concentration of protein nanoparticles by at least two different methods, including gravimetric analysis.<div><br></div></div></div>

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“…We have observed that dodecanoic acid has a significant impact on nanoparticle diameter when bound to albumin during the nanoparticle fabrication process 12 , and previous studies have shown that protein-bound fatty acids can improve albumin thermal and chemical stability 17 – 20 . It was of interest to us to determine how albumin-bound fatty acids may alter the stability of albumin nanoparticles during freeze drying and subsequent reconstitution of albumin nanoparticles.…”

Section: Introductionmentioning

confidence: 82%

Influence of bound dodecanoic acid on the reconstitution of albumin nanoparticles from a lyophilized state

Luebbert

Mansa

Rahman

et al. 2021

Sci Rep

Self Cite

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The development of reference standards for nanoparticle sizing allows for cross laboratory studies and effective transfer of particle sizing methodology. To facilitate this, these reference standards must be stable upon long-term storage. Here, we examine factors that influence the properties of cross-linked albumin nanoparticles, fabricated with an ethanol desolvation method, when reconstituted from a lyophilized state. We demonstrate, with nanoparticle tracking analysis, no significant changes in mean particle diameter upon reconstitution of albumin nanoparticles fabricated with bovine serum albumin loaded with dodecanoic acid, when compared to nanoparticles fabricated with a fatty acid-free BSA. We attribute this stability to the modulation of nanoparticle charge-charge interactions at dodecanoic acid specific binding locations. Furthermore, we demonstrate this in a lyophilized state over six months when stored at − 80 °C. We also show that the reconstitution process is readily transferable between technicians and laboratories and further confirm our finding with dynamic light scattering analysis.

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Nanoparticle size and production efficiency are affected by the presence of fatty acids during albumin nanoparticle fabrication

Cited by 13 publications

References 41 publications

Generation of BSA-capsaicin Nanoparticles and Their Hormesis Effect on the Rhodotorula mucilaginosa Yeast

Generation of BSA-capsaicin Nanoparticles and Their Hormesis Effect on the Rhodotorula mucilaginosa Yeast

Measuring the Concentration of Protein Nanoparticles Synthesized by Desolvation Method: Comparison of Bradford Assay, BCA Assay, hydrolysis/UV Spectroscopy and Gravimetric Analysis

Influence of bound dodecanoic acid on the reconstitution of albumin nanoparticles from a lyophilized state

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