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.

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

HIV-1 infected human plasma protein purification by organic and inorganic solvent v1 (protocols.io.bhyyj7xw)

protocols.io ◽  
2020 ◽  
Author(s):  
Sushanta Kumar ◽  
Deepika Varshney ◽  
Deepa Bisht ◽  
Parth Sarathi ◽  
Srikanth Prasad ◽  
...  
Keyword(s):  
Protein Purification ◽  
Human Plasma ◽  
Plasma Protein ◽  
Human Plasma Protein ◽  
Hiv 1

Use of organic solvents for protein purification from HIV-1 infected human plasma

2019 ◽  
Author(s):  
Sushanta Kumar Barik ◽  
Keshar Kunja Mohanty ◽  
Deepa Bisht ◽  
Partha Sarathi Mohanty ◽  
Shripad Patil ◽  
...  
Keyword(s):  
Protein Purification ◽  
Human Plasma ◽  
Organic Solvents ◽  
Plasma Protein ◽  
Low Cost ◽  
Laboratory Practice ◽  
Pure Protein ◽  
Chloroform Methanol ◽  
Hiv 1

Abstract Human plasma contains various high molecular and low molecular proteins like Albumin, Globulin, Lipoproteins, Glycoproteins etc. To get a pure protein of interest , the organic solvents are very useful in the plasma protein purification steps. To achieve a low cost and reliable purification, the organic solvents are best useful in laboratory practice. Tri-choloroacetic acid and Acetone removes the Albumin and Globulin while Chloroform, methanol, isopropanol removes the Glycoproteins and Lipoproteins and would give the protein of interest in HIV-1 infected human plasma.

Two-dimensional gel electrophoresis method for investigation of human plasma proteins: detection of subtle changes during filtration leukapheresis.

1982 ◽  
Vol 28 (4) ◽  
pp. 962-968 ◽  
Author(s):  
K Lonberg-Holm ◽  
E A Bagley ◽  
J Nusbacher ◽  
J M Heal
Keyword(s):  
Human Plasma ◽  
Gel Electrophoresis ◽  
Plasma Proteins ◽  
Acute Phase Proteins ◽  
Venous Blood ◽  
Sodium Dodecyl ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Native Proteins ◽  
Sds Page

Abstract Special problems are associated with analysis of human plasma proteins by standard "high-resolution" two-dimensional gel electrophoresis methods in which isoelectric focusing is followed by electrophoresis in sodium dodecyl sulfate/polyacrylamide gels (SDS-PAGE). Individual plasma proteins are often separated into overlapping groups of multiple spots, and identification of individual spots is further confounded by genetic variation. Analytical recovery of components of high molecular mass is also low or variable. These problems may be reduced or overcome by use of a "low-resolution" method consisting of electrophoresis of native proteins at pH 8.6 in an agarose gel followed by SDS-PAGE without added reducing agent. About 60 proteins can be resolved, most as single spots. About 25 of these proteins have been "mapped," and tentatively identified. We have examined 119 plasma samples taken from six donors who were undergoing filtration leukapheresis and 10 donors who were undergoing centrifugation leukapheresis or plateletpheresis. In all cases, passage of blood through a nylon filter induced a significant increase in a doublet of spots tentatively identified as complement component C3c. This was detected in the effluent from the filter throughout the first 30 min of filtration, and to a lesser extent in the venous blood. These spots were not induced by the centrifugation procedures. One filtration donor also showed increased acute-phase proteins 24 h after the procedure.

Plasma Protein, Symposia on Nutrition of the Robert Gould Research Foundation

PEDIATRICS ◽  
1951 ◽  
Vol 8 (5) ◽  
pp. 751-752
Keyword(s):  
Human Plasma ◽  
Plasma Protein ◽  
Plasma Proteins ◽  
Original Research ◽  
Edema Formation ◽  
Disease States ◽  
Serum Fractions ◽  
Normal Human ◽  
Amino Acid Antagonists ◽  
Relationship Of

This volume contains 17 separate papers or chapters reporting results of original research and discussions by recognized authorities on all important aspects of the plasma protein problem. Some of the topics discussed which are of special interest to the clinician are those relating to (1) the fractionation and properties of normal human plasma proteins, (2) plasma protein formation in health and in various disease states, (3) effects of various types of diet on plasma protein fabrication, (4) the fate of intravenously administered human plasma proteins in health, in idiopathic hypoproteinemia and in osteoporosis, (5) the mechanism of edema formation in relationship to hypoproteinemia, (6) the relationship of protein metabolism to resistance to infection, (7) the influence of the adrenal cortex on plasma protein formation and utilization and (8) the quantitative immunochemical data concerning antibody-containing serum fractions obtained by salt precipitation, alcohol precipitation or delipidation. A new approach to certain obscure metabolic disorders may, in the opinion of the reviewer, grow out of studies on amino acid antagonists discussed in the Symposium by one author.

CONFIRMATION OF PLASMA PROTEIN CHANGES IN AVIAN ERYTHROBLASTOSIS

1964 ◽  
Vol 42 (2) ◽  
pp. 293-297 ◽  
Author(s):  
M. Merriman ◽  
C. le Q. Darcel
Keyword(s):  
Plasma Protein ◽  
Gel Electrophoresis ◽  
Plasma Proteins ◽  
Starch Gel Electrophoresis ◽  
Starch Gel

Alterations of the plasma proteins have been previously demonstrated in avian erythroblastosis by paper and now by starch gel electrophoresis. With the latter technique, eight protein zones are recognized in normal plasmas. Heparin contributes an additional non-staining zone. In leukemic plasmas two more zones occur while another zone shows significant retardation.Heparin is not responsible for these changes because they are also observed in oxalated plasmas, but there is evidence of increased binding of heparin in leukemic plasma.

Identification of human plasma proteins using the Laemmli method of gel electrophoresis

1994 ◽  
Vol 22 (1) ◽  
pp. 26S-26S
Author(s):  
GEORGE J. ALLEN ◽  
BRIAN J. LONG ◽  
G. BRIAN WISDOM
Keyword(s):  
Human Plasma ◽  
Gel Electrophoresis ◽  
Plasma Proteins

scholarly journals Use of Tri-Chloroacetic Acid /Acetone/polyethylene glycol for protein purification from HIV-1 infected human plasma

2019 ◽  
Author(s):  
Sushanta Kumar Barik ◽  
Keshar Kunja Mohanty ◽  
Deepa Bisht ◽  
Partha Sarathi Mohanty ◽  
Shripad Patil ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Polyethylene Glycol ◽  
Protein Purification ◽  
Human Plasma ◽  
Water Soluble ◽  
Low Molecular Weight ◽  
High Concentration ◽  
Low Concentration ◽  
Low Molecular Weight Proteins ◽  
Hiv 1

Abstract Human plasma contains high amount of abundant proteins like albumin and globulin. Normally, the proteins having potential for biomarkers are present in very low concentration in human plasma. To resolve the low concentration proteins in polyacrylamide gel, the removal of high abundant proteins from plasma are very essential. Polyethylene glycol is a nontoxic, water soluble synthetic polymer has several applications in chemical and biomedical industries. Various molecular variants of poly ethylene glycol is available and used in protein purification. The mechanism behind the use of high concentration of polyethylene glycol is it binds the molecule in more compact or interpenetrates forming a gel like network surrounding the molecule. Polyethylene glycol -6000 removes the high abundant proteins like Albumin and Globulin in the HIV -1 infected plasma samples and concentrates the low molecular weight proteins as the low molecular weight proteins are essential in biomarker study.

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