Measuring the concentration of protein nanoparticles synthesized by desolvation method: Comparison of Bradford assay, BCA assay, hydrolysis/UV spectroscopy and gravimetric analysis

Khramtsov, Pavel; Kalashnikova, T V; Бочкова, М. С.; Kropaneva, Maria; Тимганова, В. П.; Заморина, С. А.

doi:10.1016/j.ijpharm.2021.120422

Cited by 52 publications

(31 citation statements)

References 97 publications

(2 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As we mentioned in Section our goal was to prepare relatively small nanoparticles, less than 200 nm, which is possible by using smaller gelatin concentrations for all tested gelatin types. Data obtained in the course of optimization experiments and our previous results (Khramtsov, 2021) both demonstrate that gelatin nanoparticles can be prepared at high starting gelatin concentrations. Variation of pH and volume of the desolvating agent is a possible way to decrease the size of nanoparticles when the concentration of gelatin is high.…”

Section: Discussionsupporting

confidence: 59%

“…At the same time, homogeneous suspensions of gelatin nanoparticles with polydispersity indices lower than 0.1 were formed when mixing was performed on the rotator (Figure 3). We did not perform experiments with higher gelatin concentrations, but recently we successfully prepared gelatin nanoparticles using 30 mg/ml gelatin solution which was desolvated under short gentle mixing (Khramtsov, 2021).…”

Section: Stirring Promotes Nanoparticle Aggregation In the Course Of Desolvationmentioning

confidence: 99%

“…Protein nanoparticles cross-linked with glutaraldehyde emit fluorescence when excited by UV or visible light (Cai, 2016, Khramtsov, 2021. Autofluorescence of gelatin nanoparticles can be explained by the presence of C=C (resulting from glutaraldehyde polymerization) and C=N bonds (in the Shiff bases) in nanoparticles' structure (Wei, 2007).…”

Section: Absorbance and Fluorescence Spectra Of Gelatin Nanoparticlesmentioning

confidence: 99%

“…Intensive stirring promotes gelatin aggregation. Using a simple one-step stirring-free approach we previously prepared stable and homogeneous gelatin nanoparticles from gelatin with bloom value as low as 75 (Khramtsov, 2021). Based on these findings we intended to develop a facile method for nanoparticle preparation from any type of gelatin with any bloom number.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Modified desolvation method enables simple one-step synthesis of gelatin nanoparticles from different gelatin types with any bloom values

Khramtsov

Burdina

Lazarev

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Gelatin nanoparticles found numerous applications in drug delivery, bioimaging, immunotherapy, and vaccine development as well as in biotechnology and food science. Synthesis of gelatin nanoparticles is usually made by a two-step desolvation method, which, despite providing stable and homogeneous nanoparticles, has many limitations, namely complex procedure, low yields, and poor reproducibility of the first desolvation step. Herein, we present a modified one-step desolvation method, which enables the quick, simple, and reproducible synthesis of gelatin nanoparticles. Using the proposed method one can prepare gelatin nanoparticles from any type of gelatin with any bloom number, even with the lowest ones, which remains unattainable for the traditional two-step technique. The method relies on quick one-time addition of poor solvent (preferably isopropyl alcohol) to gelatin solution in the absence of stirring. We applied the modified desolvation method to synthesize nanoparticles from porcine, bovine, and fish with bloom values from 62 to 225 on the hundreds-of-milligram scale. Synthesized nanoparticles had average diameters between 130 and 190 nm and narrow size distribution. Yields of synthesis were 62-82% and can be further increased. Gelatin nanoparticles have good colloidal stability and withstand autoclaving. Moreover, they were non-toxic to human immune cells.

show abstract

Section: Discussionsupporting

confidence: 59%

Section: Stirring Promotes Nanoparticle Aggregation In the Course Of Desolvationmentioning

confidence: 99%

Section: Absorbance and Fluorescence Spectra Of Gelatin Nanoparticlesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Modified desolvation method enables simple one-step synthesis of gelatin nanoparticles from different gelatin types with any bloom values

Khramtsov

Burdina

Lazarev

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…SDS-PAGE was used to additionally characterize the structure of BSA/Hemin nanoparticles. Recently, we demonstrated that disulfide bonds maintain heat-stabilized BSA nanoparticle integrity [Khramtsov, 2021]. Nanoparticles were treated with betamercaptoethanol which is able to reduce protein disulfide bonds and applied to the gel.…”

Section: Figure 3 Sem Images Of Bsa/hemin Nanoparticles: Np5 (A) Np10 (B) Np20 (C) Np40 (D) Np80 (E) Npc (F) Scale Bar Is 300 Nmmentioning

confidence: 99%

Albumin nanoparticles loaded with hemin as peroxidase mimics for immunoassay

Khramtsov

Бочкова

Тимганова

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Contemporary immunoassays commonly used in clinical diagnostics mostly utilize enzymes, such as horseradish peroxidase, for signal generation. Numerous research is dedicated to the development of artificial peroxidase-mimicking catalysts with lower cost, high activity, better operational stability, and tunable properties. Herein we synthesized hemin-loaded bovine serum albumin (BSA) nanoparticles and applied them as catalytic labels (nanozymes) in colorimetric immunoassay of anti-tetanus antibodies. Hemin is a key part of the peroxidase catalytic center, possessing peroxidase like-activity. Albumin nanoparticles were loaded with multiple hemin molecules and decorated with Streptococcal protein G. Resulting nanozymes possessed good colloidal stability and allowed for antibody detection in blood serum. The sensitivity of antibody detection was sufficient for the assessment of post-vaccination immunity.

show abstract

Improvement of an enzymatic cascade synthesis of nicotinamide mononucleotide via protein engineering and reaction‐process reinforcement

Peng,

Hong,

Cui

et al. 2024

Biotechnology Journal

View full text Add to dashboard Cite

Enzymatic synthesis of β‐nicotinamide mononucleotide (NMN) from D‐ribose has garnered widespread attention due to its cheap material, the use of mild reaction conditions, and the ability to produce highly pure products with the desired optical properties. However, the overall NMN yield of this method is impeded by the low activity of rate‐limiting enzymes. The ribose‐phosphate diphosphokinase (PRS) and nicotinamide phosphoribosyltransferase (NAMPT), that control the rate of the reaction, were engineered to improve the reaction efficacy. The actives of mutants PRS‐H150Q and NAMPT‐Y15S were 334% and 57% higher than that of their corresponding wild‐type enzymes, respectively. Furthermore, by adding pyrophosphatase, the byproduct pyrophosphate which can inhibit the activity of NAMPT was degraded, leading to a 6.72% increase in NMN yield. Following with reaction‐process reinforcement, a high yield of 8.10 g L−1 NMN was obtained after 3 h of reaction, which was 56.86‐fold higher than that of the stepwise reaction synthesis (0.14 g L−1), indicating that the in vitro enzymatic synthesis of NMN from D‐ribose and niacinamide is an economical and feasible route.

show abstract

Measuring the concentration of protein nanoparticles synthesized by desolvation method: Comparison of Bradford assay, BCA assay, hydrolysis/UV spectroscopy and gravimetric analysis

Cited by 52 publications

References 97 publications

Modified desolvation method enables simple one-step synthesis of gelatin nanoparticles from different gelatin types with any bloom values

Modified desolvation method enables simple one-step synthesis of gelatin nanoparticles from different gelatin types with any bloom values

Albumin nanoparticles loaded with hemin as peroxidase mimics for immunoassay

Improvement of an enzymatic cascade synthesis of nicotinamide mononucleotide via protein engineering and reaction‐process reinforcement

Contact Info

Product

Resources

About