scholarly journals Identification of Abnormal Proteins in Plasma from Gout Patients by LC-MS/MS

Separations ◽  
2021 ◽  
Vol 8 (6) ◽  
pp. 85
Author(s):  
Lijin Shen ◽  
Hanyang Dong ◽  
Zhenchang Guo ◽  
Guijin Zhai ◽  
Kai Zhang
Keyword(s):  
Human Plasma ◽  
Plasma Proteins ◽  
High Performance ◽  
Clinical Analysis ◽  
Kidney Damage ◽  
Human Blood Plasma ◽  
Protein Levels ◽  
Differential Proteins ◽  
High Level ◽  
Abundance Proteins

A high level of uric acid may cause hyperuricemia, which further develops into gout, eventually leading to chronic kidney disease. However, the pathogenic mechanism remains largely unknown. To investigate the cause and block the transformation of hyperuricemia to related diseases, it is important to discover the alterations in protein levels between gout patients and non-gout individuals. To date, human blood plasma is still the predominant matrices for clinical analysis. Due to the high abundance, the proteins of plasma samples have strong shielding effects on low abundance proteins, thus, the information on low abundance protein expression is always masked, while the low abundance proteins of human plasma are often of great significance for the diagnosis and treatment of diseases. Therefore, it is very important to separate and analyze the plasma proteins. High-performance liquid chromatography (LC) tandem mass spectrometry (MS)-based proteomics has been developed as a powerful tool to investigate changes in the human plasma proteome. Here, we used LC-MS/MS to detect the differential proteins in the plasmas from simple gout patients, gout with kidney damage patients, and non-gout individuals. We identified 32 obviously differential proteins between non-gout and gout subjects and 10 differential proteins between simple gout and gout with kidney damage patients. These differential proteins were further analyzed to characterize their localization and functions. Additionally, the correlation analysis showed multiple relationships between the abnormal plasma proteins and clinical biochemical indexes, particularly for the immune-inflammatory response proteins. Furthermore, inflammation factors gelsolin (GSN) were confirmed. Our results offer a view of plasma proteins for studying biomarkers of gout patients.

Purification of glycopeptides of human plasma proteins by high-performance liquid chromatography

1984 ◽  
Vol 317 ◽  
pp. 11-26 ◽  
Author(s):  
Nobuhiro Takahashi ◽  
Yoko Takahashi ◽  
Thomas L. Ortel ◽  
Jay N. Lozier ◽  
Noriaki Ishioka ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Human Plasma ◽  
Plasma Proteins ◽  
High Performance

scholarly journals A Recombinant Protein Biomarker DDA Library Increases DIA Coverage of Low Abundance Plasma Proteins

2020 ◽  
Author(s):  
Seong Beom Ahn ◽  
Karthik S. Kamath ◽  
Abidali Mohamedali ◽  
Zainab Noor ◽  
Jemma X. Wu ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Human Plasma ◽  
Recombinant Proteins ◽  
Plasma Proteins ◽  
Biomarker Discovery ◽  
Cancer Biomarkers ◽  
Human Blood Plasma ◽  
Spectral Library ◽  
Protein Inference ◽  
High Stringency

AbstractCredible detection and quantification of low abundance proteins from human blood plasma is a major challenge in precision medicine biomarker discovery when using mass spectrometry (MS). Here, we employed a mixture of recombinant proteins in DDA libraries to subsequently detect cancer-associated low abundance plasma proteins using SWATH/DIA. The exemplar DDA recombinant protein spectral library (rPSL) was derived from tryptic digestion of 36 human recombinant proteins that had been previously implicated as possible cancer biomarkers in both our own and other studies. The rPSL was then used to identify proteins from non-depleted colorectal cancer (CRC) plasmas by SWATH-MS. Most (32/36) of the proteins in the rPSL were reliably identified in plasma samples, including 8 proteins (BTC, CXCL10, IL1B, IL6, ITGB6, TGFα, TNF, TP53) not previously detected using high-stringency MS in human plasmas according to PeptideAtlas. The rPSL SWATH-MS protocol was compared to DDA-MS using MARS-depleted and post-digestion peptide fractionated plasmas (here referred to as a human plasma DDA library). Of the 32 proteins identified using rPSL SWATH, only 12 were identified using DDA-MS. The 20 additional proteins exclusively identified by using the rPSL approach with SWATH were mostly lower abundance (i.e., <10ng/ml) plasma proteins. To mitigate FDR concerns, and replicating a more typical approach, the DDA rPSL was also merged into a human plasma DDA library. When SWATH identification was repeated using this merged library, the majority (33/36) of low abundance plasma proteins from the rPSL could still be identified using high-stringency HPP Guidelines v3.0 protein inference criteria.

Comparative evaluation of plasma protein purification and 2-D gel electrophoresis protocols for analysis of HIV-1 infected plasma proteins (Preprint)

2021 ◽  
Author(s):  
Mr Sushanta Kumar Barik Sr ◽  
Deepika Varshney ◽  
Deepa Bisht Sr ◽  
Shripad A Patil Sr ◽  
Rananjaya Singh Sr ◽  
...  
Keyword(s):  
Protein Purification ◽  
Human Plasma ◽  
Plasma Protein ◽  
Gel Electrophoresis ◽  
Comparative Evaluation ◽  
Plasma Proteins ◽  
Human Plasma Protein ◽  
Sds Page ◽  
Hiv 1 ◽  
Abundance Proteins

BACKGROUND The focus of the study was the comparative evaluation of HIV-1 infected plasma protein purification and 2-D gel electrophoresis protocols. OBJECTIVE Human plasma protein purification is a risk task to perform 2-D gel electrophoresis. Human plasma proteins contain nearly 70% albumin and globulin. The removal of such high abundance high molecular weight proteins is very difficult to perform 2-D gel electrophoresis METHODS To the best of our knowledge, we searched several research papers, developed and adopted various organic and non-organic based protocols for HIV-1 infected human plasma protein purification and various isoelectrofocusing protocols in 2-D gel electrophoresis RESULTS After failure in 2-D gel-electrophoresis performance by these protocols, Aurum serum mini kit (Bio-Rad, USA) was adopted for plasma protein purification for performing 2-D gel electrophoresis. The low-abundance proteins were better resolved by 10% SDS-PAGE in 2-D gel-electrophoresis. Then, we extended the MALDI-TOF/TOF analysis of low-abundance proteins in human plasma by adopting the Aurum serum mini kit ( Bio-Rad, USA) for 2-D gel electrophoresis. CONCLUSIONS Thus, we concluded that, the Aurum serum mini kit (Bio-Rad, USA) is best to perform the 2-D gel electrophoresis of HIV-1 infected human plasma by depleting the high abundant proteins like albumin and globulin

scholarly journals Preprint: A robust peptidomics mass spectrometry platform for measuring oxytocin in plasma and serum

2016 ◽  
Author(s):  
Ole Kristian Brandtzaeg ◽  
Elin Johnsen ◽  
Hanne Roberg-Larsen ◽  
Knut Fredrik Seip ◽  
Siri Leknes ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Human Plasma ◽  
Plasma Proteins ◽  
High Performance ◽  
Cyclic Peptide ◽  
Liquid Chromatography Mass Spectrometry ◽  
Cord Serum ◽  
Chromatography Mass Spectrometry ◽  
Nano Liquid Chromatography

Current approaches to measuring the cyclic peptide oxytocin in plasma/serum are associated with poor selectivity and/or inadequate sensitivity. We here describe a high performance nano liquid chromatography-mass spectrometry platform for measuring OT in human plasma/serum. The platform is extremely robust, allowing laborious sample clean-up steps to be omitted. OT binds strongly to plasma proteins, but a reduction/alkylation procedure breaks this bond, allowing ample detection of total OT. The method showed excellent quantitation properties, and was used to determine total OT levels to 0.5-1.2 ng/mL (evaluated with human plasma and cord serum). The method is compatible with accessible mass spectrometry instrumentation, finally allowing selective and easily comparable oxytocin measurements.

scholarly journals Development and Validation of Pomalidomide Determination in Human Plasma by HPLC-MS/MS Method

2020 ◽  
Vol 9 (4) ◽  
pp. 146-154
Author(s):  
T. N. Komarov ◽  
I. E. Shohin ◽  
M. A. Tokareva ◽  
O. A. Archakova ◽  
D. S. Bogdanova ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Blood Plasma ◽  
Human Plasma ◽  
High Performance ◽  
Protein Precipitation ◽  
Human Blood Plasma ◽  
Pharmacokinetic Studies ◽  
Limit Of Quantification ◽  
Development And Validation

Introduction. B-cell malignancies of the plasma cell leads to the second most spread hematological malignancy disease, called multiple myeloma. Pomalidomide is used in case of previous multiple myeloma ineffective treatment. Pomalidomide is a thalidomide synthetic derived, approved as immunomodulatory drug by the Food and Drug Administration (FDA). Nowadays, detection of pomalidomide in blood plasma by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) is not spread. Moreover, the detection and the experimental setting accumulated data are varying greatly. This investigation provides development and validation of pomalidomide aiming to determine human blood plasma by HPLC-MS/MS method. The samples were processed by methanol protein precipitation.Aim. The aim of this study is to develop a method for the pomalidomide in human plasma by HPLC-MS/MS for pharmacokinetic studies.Materials and methods. Determination of pomalidomide in plasma by HPLC-MS/MS. The samples were processed by methanol protein precipitation.Results and discussion. This method was validated by next parameters: selectivity, matrix effect, calibration curve, accuracy, precision, spike recovery, lower limit of quantification, detection limit, carry-over and stability. Conclusion. The method of the determination of pomalidomide in human plasma was developed and validated by HPLC-MS/MS. The linearity in plasma sample was achieved in the concentration range of 1,00 – 500,00 ng/ml. Method could be applied to pomalidomide determination in plasma for PK and BE studies.

scholarly journals Comparative evaluation of plasma protein purification and 2-D gel electrophoresis protocols for analysis of HIV-1 infected human plasma proteins

2021 ◽  
Author(s):  
Sushanta Kumar Barik ◽  
Deepika Varshney ◽  
Keshar Kunja Mohanty ◽  
Deepa Bisht ◽  
Shripad A. Patil ◽  
...  
Keyword(s):  
Protein Purification ◽  
Human Plasma ◽  
Plasma Protein ◽  
Gel Electrophoresis ◽  
Comparative Evaluation ◽  
Plasma Proteins ◽  
Sds Page ◽  
High Molecular Weight Proteins ◽  
Hiv 1 ◽  
Abundance Proteins

Abstract Purification of proteins from human plasma is a herculean task to perform 2-D gel electrophoresis. Human plasma contains nearly 70% albumin and globulin. The removal of such high abundance high molecular weight proteins is very difficult before performing 2-D gel electrophoresis. It becomes more difficult when we intent to investigate in infectious diseases like HIV/AIDS. We tried to the best of our efforts adopting various organic and non-organic based protocols based on various published papers. After failure of these protocols in results of 2-D gel-electrophoresis Aurum serum mini kit (Bio-Rad, USA) was adopted for plasma protein purification for performing 2-D gel electrophoresis. The low-abundance proteins were better resolved by 10% SDS-PAGE in 2-D gel-electrophoresis. Then,we extended the MALDI-TOF/TOF analysis of low-abundance proteins in human plasma by adopting the Aurum serum mini kit (Bio-Rad, USA) for 2-D gel electrophoresis. Thus, we concluded that, depletion of high abundant proteins like albumin and globulin, the use of the Aurum serum mini kit (Bio-Rad, USA) is the protocol of choice to perform the 2-D gel electrophoresis of HIV-1 infected human plasma.

scholarly journals Analysis of Native Human Plasma Proteins and Haemoglobin for the Presence of Bityrosine by High-Performance Liquid Chromatography

1997 ◽  
Vol 81 (5) ◽  
pp. 205-208 ◽  
Author(s):  
Bahrain Daneshvar ◽  
Henrik Frandsen ◽  
Lars O. Dragsted ◽  
Lisbeth E. Knudsen ◽  
Herman Autrup
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Human Plasma ◽  
Plasma Proteins ◽  
High Performance

scholarly journals Development and Validation of Tranexamic Acid Determination in Human Plasma by HPLC-MS/MS method

2021 ◽  
Vol 10 (2) ◽  
pp. 120-127
Author(s):  
A. V. Aleshina ◽  
T. N. Komarov ◽  
O. A. Archakova ◽  
D. S. Shchelgacheva ◽  
N. S. Bagaeva ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Tranexamic Acid ◽  
Human Plasma ◽  
Analytical Method ◽  
High Performance ◽  
Human Blood Plasma ◽  
Pharmacokinetic Studies ◽  
Acid Determination

Introduction. Tranexamic acid is one of the most common drugs used to stop bleeding after trauma, in surgery and gynecology. The most common analytical method for the determination of this compound is reversed-phase high-performance liquid chromatography (HPLC). However, this compound belongs to the group of so-called poorly retained compounds due to its chemical structure. It is necessary to develop an analytical method that will allow the determination of tranexamic acid in human blood plasma with the least time, resource costs and without the use of specialized columns.Aim. The aim of this study is to develop a method for tranexamic acid in human plasma by high performance liquid chromatography with tandem mass-spectrometry (HPLC-MS/MS) for pharmacokinetic studies.Materials and methods. Determination of tranexamic acid in plasma by HPLC-MS/MS. The samples were processed by acetonitrile protein precipitation.Results and discussion. This method was validated by next parameters: selectivity, matrix effect, calibration curve, accuracy, precision, recovery, lower limit of quantification, carry-over effect and stability.Conclusion. The method of the determination of tranexamic acid in human plasma was developed and validated by HPLC-MS/MS. The linearity in plasma sample was achieved in the concentration range of 100.00–15000.00 ng/ml. Method could be applied to tranexamic acid determination in plasma for pharmacokinetics and bioequivalence studies.

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