Application of tandem fast protein liquid chromatography to purify intact native monomeric/aggregated Tamm–Horsfall protein from human urine and systematic comparisons with diatomaceous earth adsorption and salt precipitation: yield, purity and time-consumption

Abstract

Tamm-Horsfall protein (THP) is a high-abundance urinary protein. Although its functions have been studied for years, several aspects of those remain unclear. To achieve more knowledge on THP functions, effective...

Contamination of bacterial extracellular vesicles (bEVs) in human urinary extracellular vesicles (uEVs) samples and their effects on uEVs study

“…The nanoscale uEVs derived from 30‐ml aliquot of the basal urine without storage nor NaN 3 were extracted using Laemmli's buffer, whereas the whole basal urine (30‐ml) without storage nor NaN 3 was subjected to dialysis against deionized water and concentrated by lyophilization as previously described (Noonin et al., 2021 , 2022 ). Protein concentration in each sample was measured by Bradford's method using Bio‐Rad Protein Assay (Bio‐Rad Laboratories; Hercules, CA).…”

Fast Protein and Metabolites (Nucleotides and Nucleosides) Liquid Chromatography Technique and Chemical Sensor for the Assessment of Fish and Meat Freshness

“…The positive features of FPLC include high loading capacity, biocompatible aqueous buffer systems, high flow rates, and the availability of stationary phases for most common chromatographic modes (e.g., ion exchange, gel filtration, reversed phase, and affinity chromatography) [4]. The use of FPLC as a characterization and analytical technique can be particularly relevant in the fields where the availability of rapid and affordable analytical methods and tools is crucial, e.g., routine test in healthcare or the food industry [2,3,[5][6][7][8][9]. In this context, FPLC can be modified and enhanced with the ability to fractionate and detect low and middle molecular weight metabolites: ATP metabolites nucleotides and nucleosides, advanced glycation end products (AGE), uric acid, and other metabolic products originating from various biological samples [5,[7][8][9].…”

“…The use of FPLC as a characterization and analytical technique can be particularly relevant in the fields where the availability of rapid and affordable analytical methods and tools is crucial, e.g., routine test in healthcare or the food industry [2,3,[5][6][7][8][9]. In this context, FPLC can be modified and enhanced with the ability to fractionate and detect low and middle molecular weight metabolites: ATP metabolites nucleotides and nucleosides, advanced glycation end products (AGE), uric acid, and other metabolic products originating from various biological samples [5,[7][8][9]. In our opinion, the term fast protein and metabolites liquid chromatography (FPMLC) is more appropriate for such applications and will continue be used in this paper.…”