scholarly journals Comprehensive Profiling of Plasma Exosomes Using Data-Independent Acquisitions – New Tools for Aging Cohort Studies

2021 ◽  
Author(s):  
Sandip K. Patel ◽  
Roland Bruderer ◽  
Nathan Basisty ◽  
Joanna Bons ◽  
Pierre-Yves Desprez ◽  
...  
Keyword(s):  
Reversed Phase ◽  
The Body ◽  
Size Exclusion ◽  
Biomarkers Of Aging ◽  
Age Related ◽  
Complex Biological Process ◽  
High Throughput Method ◽  
Aging Study ◽  
Exclusion Chromatography ◽  
Using Data

AbstractAging is a complex biological process associated with progressive loss of physiological function and susceptibility to several diseases, such as cancer and neurodegeneration. Exosomes are involved in many cellular signaling pathways, and their cargo may serve as promising disease or aging biomarkers. These membrane-bound extracellular vesicles facilitate the transport of intracellular contents to proximal and distal cells in the body. Here, we investigated two omics approaches for exosome analysis. To overcome the challenges of plasma exosome contamination with abundant soluble plasma proteins, we developed a high-throughput method to isolate highly purified exosomes from human plasma by sequential size-exclusion chromatography and ultrafiltration. First, we used data-dependent acquisitions from offline high-pH reversed-phase fractions of exosome lysate to generate a deep spectral library comprising ∼2,300 exosome proteins. Second, in a pilot aging study, we used comprehensive data-independent acquisitions to compare plasma exosomes from young (20–26 yrs) and old (60–66 yrs) individuals. We quantified 1,318 exosome proteins, and levels of 144 proteins were significantly different in young and old plasma groups (Q<0.05 and >1.5-fold change). We also analyzed exosome miRNA cargo and detected 331 miRNAs. Levels of several were significantly different in young and old individuals. In addition, 88 and 17 miRNAs were unique to old and young individuals, respectively. Plasma exosome biomarkers have great potential for translational studies investigating biomarkers of aging and age-related diseases and to monitor therapeutic aging interventions.

scholarly journals Identification of food-derived peptides in human blood after ingestion of corn and wheat gluten hydrolysates

2018 ◽  
Vol 2 ◽  
Author(s):  
Akika Ejima ◽  
Megumi Nakamura ◽  
Yasushi A. Suzuki ◽  
Kenji Sato
Keyword(s):  
Matrix Protein ◽  
Leucine Aminopeptidase ◽  
Wheat Gluten ◽  
Protein Hydrolysates ◽  
The Body ◽  
Size Exclusion ◽  
Extracellular Matrix Protein ◽  
Reverse Phase Hplc ◽  
Before And After ◽  
Exclusion Chromatography

Bioactive peptides in the body after ingestion of plant protein hydrolysates have been speculated but not identified. We aimed to establish an approach to identify small amounts of food-derived peptides in humans after ingestion of non-extracellular matrix protein hydrolysates. Corn and wheat gluten hydrolysates were digested using pancreatin and leucine aminopeptidase; the resultant peptides were identified via size-exclusion chromatography and reverse-phase HPLC-tandem mass spectrometry (MS/MS). Structures of indigestible peptides were confirmed via LC-MS/MS in multi-reaction monitoring mode. All indigestible peptides in the exopeptidase digest were diprolyl and di- and tripyroglutamyl peptides. Blood collected from healthy volunteers (n = 4) before and after ingestion of 9 g of the hydrolysates was assessed for the indigestible peptides via LC-MS/MS. Six peptides (Pro-Ala, Pro-Gly, Pro-Gln, pyroGlu-Pro, pyroGlu-Leu-Pro, and pyroGlu-Gln-Pro) significantly increased in human plasma up to 10–100 nM compared to the baseline. This may hence be a powerful tool for identifying foodderived peptides in blood.

scholarly journals BIOLOGICAL AGE AS A METHOD FOR ESTIMATING HEALTH LEVEL UNDER ENVIRONMENTAL RISKS (LITERATURE REVIEW)

2019 ◽  
Vol 98 (7) ◽  
pp. 761-765 ◽  
Author(s):  
N. I. Prokhorov ◽  
V. I. Dontsov ◽  
Vyacheslav N. Krutko ◽  
T. M. Khodykina
Keyword(s):  
Human Body ◽  
Quantitative Assessment ◽  
Modern Human ◽  
The Body ◽  
Biological Age ◽  
Whole Body ◽  
New Methods ◽  
Biomarkers Of Aging ◽  
Age Related ◽  
Definition Of

The widespread formation of unfavorable environmental, the swiftness of modern life with large information and psycho-emotional loads and extremely natural and climatic cataclysms, as well as harmful addictions and wrong way of life of modern human, lead to the development of stress and disruption of the mechanisms of adaptation of the human body and its accelerated wear. This stimulates the development of research on the creation of new methods of integrated assessment of health and quantitative assessment of the aging processes of the body systems and the whole body, as well as the possibilities of new methods of risk assessment of climatic and environmentally related pathological and age-related diseases. The aim of the work was to consider the methodology of quantitative assessment of individual health and the rate of aging of the human body on the basis of the system index of Biological age (BA); description of its essence and structure, requirements for tests - biomarkers of aging used as the index of BA, definition of possibilities and scope of application of the BA method in modern practice of Biomedicine. The use of modern methods of scientific analysis - a systematic approach to the analysis of the processes of human aging and determine its quantitative side - the value of BA, allows a reasonable approach to the choice of the number of BM, to take into account their information content and precision, and the cost of diagnostics and availability for different users, to take into account the specific objectives of the researcher. The use of the index-partial BA allows individual approaching the choice of biomarkers and create personalized panels for the definition of BA programs for the prevention of aging in personalized preventive medicine. The complexity of the content and calculation of indices of BA requires automation and the use of methods of modern computer science and computer calculations and programs. For this purpose, we have created special computer software for diagnosing aging by calculating the BA indices with the possibility of choosing BM and automatic calculation of indicators and conclusions.

scholarly journals Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays

Toxins ◽  
2019 ◽  
Vol 11 (1) ◽  
pp. 35 ◽  
Author(s):  
Bruna Xavier ◽  
Rafaela Perobelli ◽  
Maurício Walter ◽  
Francielle da Silva ◽  
Sérgio Dalmora
Keyword(s):  
Botulinum Neurotoxin ◽  
Type A ◽  
Reversed Phase ◽  
Size Exclusion ◽  
Mouse Bioassay ◽  
Forced Degradation ◽  
Mean Values ◽  
Botulinum Neurotoxin Type A ◽  
Exclusion Chromatography ◽  
Cell Culture Assay

Botulinum neurotoxin type-A (BoNTA) is one of the seven different serotypes (A to G) produced by Clostridium botulinum. A stability-indicating size-exclusion chromatography (SEC) method was developed and validated, and the specificity was confirmed by forced degradation study, interference of the excipients, and peaks purity. The method was applied to assess the content and high-molecular-weight (HMW) forms of BoNTA in biopharmaceutical products, and the results were compared with those of the LD50 mouse bioassay, the T−47D cell culture assay, and the reversed-phase chromatography (RPC) method, giving mean values of 0.71% higher, 0.36% lower, and 0.87% higher, respectively. Aggregated forms showed significant effects on cytotoxicity, as well as a decrease in the bioactivity (p < 0.05). The employment of the proposed method in conjunction with the optimized analytical technologies for the analysis of the intact and altered forms of the biotechnology-derived medicines, in the correlation studies, enabled the demonstration of the capability of each one of the methods and allowed for great improvements, thereby assuring their safe and effective use.

Separation of Liquiritin by Two-Dimensional Liquid Chromatography

2012 ◽  
Vol 455-456 ◽  
pp. 1232-1238 ◽  
Author(s):  
Jing Xiang Cong ◽  
Shao Yan Wang ◽  
Hong Gao
Keyword(s):  
Liquid Chromatography ◽  
Reversed Phase ◽  
Normal Phase ◽  
Size Exclusion ◽  
Two Dimensional ◽  
Target Compound ◽  
Reversed Phase Chromatography ◽  
Complex Samples ◽  
Exclusion Chromatography ◽  
Licorice Extract

Two-dimensional liquid chromatography (2DLC) is an important technology for the separation and analysis of complex samples. Liquiritin, an important active component in licorice, was chosen as the target compound and it was separated by three kinds of off-line 2DLC, i.e. size exclusion chromatography × reversed phase chromatography, normal phase × reversed phase chromatography and reversed phase chromatography × reversed phase chromatography (SEC×RP, NP×RP and RP×RP). The chromatographic conditions were selected and the 2D systems were combined. The results show that it is feasible to separate Liquiritin from licorice extract using 2DLC. Among the 2D modes mentioned above, the highest purity of Liquiritin was obtained in the RP×RP mode, and the concentration of Liquiritin was increased most significantly in the NP×RP mode.

scholarly journals Coupling of Size-Exclusion Chromatography to Liquid Chromatography/Mass Spectrometry for Determination of Trace Levels of Thifensulfuron-Methyl and Tribenuron-Methyl in Cottonseed and Cotton Gin Trash

2000 ◽  
Vol 83 (3) ◽  
pp. 651-659 ◽  
Author(s):  
James J Stry ◽  
Jennifer S Amoo ◽  
Steve W George ◽  
Teresa Hamilton-Johnson ◽  
Ellen Stetser
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Size Exclusion Chromatography ◽  
Reversed Phase ◽  
Size Exclusion ◽  
Liquid Chromatography Mass Spectrometry ◽  
Exclusion Chromatography ◽  
Tribenuron Methyl ◽  
Cotton Gin

Abstract Size-exclusion chromatography (SEC) was coupled to reversed-phase liquid chromatography/mass spectrometry for the determination of thifensulfuron-methyl and tribenuron-methyl in cottonseed and cotton gin trash. The limit of quantitation was 20 parts per billion (ppb), and the limit of detection was 6 ppb. The analytes were extracted by homogenization in a buffer solution. The extracts underwent a solvent exchange into methanol and were injected onto an SEC column. As the analytes eluted from the SEC column, the eluate was diverted onto a reversed-phase column for additional separation of the analytes and their detection via mass spectrometry. This method is unique because the samples are not cleaned up before analysis, the analytes are injected in methanol, and the entire analysis is completed in 30 min. Average recoveries and standard deviations for thifensulfuronmethyl and tribenuron-methyl in cotton gin trash were 91 ± 6% and 88 ± 5%, respectively. Average recoveries and standard deviations for thifensulfuron-methyl and tribenuron-methyl in cottonseed were 91 ± 11% and 99 ± 12%, respectively. This is an effective method for the detection and determination of thifensulfuron-methyl and tribenuronmethyl in cotton.

scholarly journals Salt-free fractionation of complex isomeric mixtures of glycosaminoglycan oligosaccharides compatible with ESI-MS and microarray analysis

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Hao Liu ◽  
Apoorva Joshi ◽  
Pradeep Chopra ◽  
Lin Liu ◽  
Geert-Jan Boons ◽  
...  
Keyword(s):  
Structure Function ◽  
Structural Complexity ◽  
Reversed Phase ◽  
Separation Method ◽  
Size Exclusion ◽  
Reaction Products ◽  
Exclusion Chromatography ◽  
Function Characterization ◽  
Function Relationship ◽  
Isomeric Mixtures

Abstract Heparin and heparan sulfate (Hp/HS) are linear complex glycosaminoglycans which are involved in diverse biological processes. The structural complexity brings difficulties in separation, making the study of structure-function relationships challenging. Here we present a separation method for Hp/HS oligosaccharide fractionation with cross-compatible solvent and conditions, combining size exclusion chromatography (SEC), ion-pair reversed phase chromatography (IPRP), and hydrophilic interaction chromatography (HILIC) as three orthogonal separation methods that do not require desalting or extensive sample handling. With this method, the final eluent is suitable for structure-function relationship studies, including tandem mass spectrometry and microarray printing. Our data indicate that high resolution is achieved on both IPRP and HILIC for Hp/HS isomers. In addition, the fractions co-eluted in IPRP could be further separated by HILIC, with both separation dimensions capable of resolving some isomeric oligosaccharides. We demonstrate this method using both unpurified reaction products from isomeric synthetic hexasaccharides and an octasaccharide fraction from enoxaparin, identifying isomers resolved by this multi-dimensional separation method. We demonstrate both structural analysis by MS, as well as functional analysis by microarray printing and screening using a prototypical Hp/HS binding protein: basic-fibroblast growth factor (FGF2). Collectively, this method provides a strategy for efficient Hp/HS structure-function characterization.

scholarly journals Characterization, Preparation, and Purification of Marine Bioactive Peptides

2017 ◽  
Vol 2017 ◽  
pp. 1-16 ◽  
Author(s):  
Xueqin Wang ◽  
Huahua Yu ◽  
Ronge Xing ◽  
Pengcheng Li
Keyword(s):  
High Performance ◽  
Bioactive Peptides ◽  
Reversed Phase ◽  
Size Exclusion ◽  
Antihypertensive Activity ◽  
Microwave Assisted ◽  
Chemical Hydrolysis ◽  
Assisted Extraction ◽  
Exclusion Chromatography ◽  
Manufacturing Techniques

Marine bioactive peptides, as a source of unique bioactive compounds, are the focus of current research. They exert various biological roles, some of the most crucial of which are antioxidant activity, antimicrobial activity, anticancer activity, antihypertensive activity, anti-inflammatory activity, and so forth, and specific characteristics of the bioactivities are described. This review also describes various manufacturing techniques for marine bioactive peptides using organic synthesis, microwave assisted extraction, chemical hydrolysis, and enzymes hydrolysis. Finally, purification of marine bioactive peptides is described, including gel or size exclusion chromatography, ion-exchange column chromatography, and reversed-phase high-performance liquid chromatography, which are aimed at finding a fast, simple, and effective method to obtain the target peptides.

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