The economic significance of honey production is crucial; therefore, modern and efficient methods of authentication are needed. During the last decade, various data processing methods and a combination of several instrumental methods have been increasingly used in food analysis. In this study, the chemical composition of monofloral buckwheat (Fagopyrum esculentum), clover (Trifolium repens), heather (Calluna vulgaris), linden (Tilia cordata), rapeseed (Brassica napus), willow (Salix cinerea), and polyfloral honey samples of Latvian origin were investigated using several instrumental analysis methods. The data from light stable isotope ratio mass spectrometry (IRMS), ultra-high performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS), and nuclear magnetic resonance (NMR) analysis methods were used in combination with multivariate analysis to characterize honey samples originating from Latvia. Results were processed using the principal component analysis (PCA) to study the potential possibilities of evaluating the differences between honey of different floral origins. The results indicate the possibility of strong differentiation of heather and buckwheat honeys, and minor differentiation of linden honey from polyfloral honey types. The main indicators include depleted δ15N values for heather honey protein, elevated concentration levels of rutin for buckwheat honey, and qualitative presence of specific biomarkers within NMR for linden honey.
Site-specific protein modifications are vital for biopharmaceutical drug development. Gluconoylation is a nonenzymatic, post-translational modification of N-terminal HisTags. We report high-yield, site-selective in vitro α-aminoacylation of peptides, glycoproteins, antibodies, and Virus-like particles (VLPs) with azidogluconolactone at pH 7.5 in 1 h. Conjugates slowly hydrolyse, but diol-masking with borate esters inhibits reversibility. In an example, we multimerise azidogluconoylated SARS-CoV-2 receptor-binding domain (RBD) onto VLPs via click-chemistry, to give a COVID-19 vaccine. Compared to yeast antigen, HEK-derived RBD was immunologically superior, likely due to observed differences in glycosylation. We show the benefits of ordered over randomly oriented multimeric antigen display, by demonstrating single-shot seroconversion and best virus-neutralizing antibodies. Azidogluconoylation is simple, fast and robust chemistry, and should accelerate research and development.
The protonation process of tyrosine kinase inhibitor lapatinib was studied by means of 1HNMR and UV/Vis spectroscopy joint with the theoretical calculations at DFT and semi-empirical levels. DFT/M06-2X geometries were used to describe and compare the different cationic forms of lapatinib, while ZINDO/S-CI method performed on those geometries allowed for the interpretation of experimental UV/Vis spectra of lapatinib at various pH. We found that at low pH two different dicationic forms (N2N1 and N1N3) of lapatinib were present in ethanol and DMSO-d6 solutions. The first protonation, however, occurred on the aliphatic N1 in DMSO-d6, while in ethanol solutions most probably the quinazoline nitrogen atom N2 was also protonated.
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