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>
Prussian blue nanozymes possessing peroxidase-like activity gather significant attention as alternatives to natural enzymes in therapy, biosensing, and environmental remediation. Recently, Prussian blue nanoparticles with enhanced catalytic activity prepared by reduction of FeCl3/K3[Fe(CN)6] mixture have been reported. These nanoparticles were denoted as ‘artificial peroxidase’ nanozymes. Our study provides insights into the process of their synthesis. We studied how the size of nanozymes and synthesis yield can be controlled via adjustment of the synthesis conditions. Based on these results, we developed a reproducible and scalable method for the preparation of ‘artificial peroxidase’ with tunable sizes and enhanced catalytic activity. Nanozymes modified with gelatin shell and functionalized with affine molecules were applied as labels in colorimetric immunoassays of prostate-specific antigen and tetanus antibodies, enabling detection of these analytes in the range of clinically relevant concentrations. Protein coating provides excellent colloidal stability of nanozymes in physiological conditions and stability upon long-term storage.
The effect of native α-fetoprotein (AFP) on the expression of T-regulatory lymphocyte (Treg) markers by activated CD4 lymphocytes with different proliferative status was studied. α-Fetoprotein did not affect the ratio of proliferating and non-proliferating activated CD4 cells. In the study of Treg differentiation, it was found that AFP at concentrations of 50 and 100 μg/mL significantly inhibited the number of nonproliferating CD4FOXP3 and CD4FOXP3HELIOS lymphocytes without affecting the expression of Treg markers by proliferating CD4 lymphocytes.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.