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

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

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

10.21203/rs.2.12144/v1 ◽

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.

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

10.1101/2021.03.03.433830 ◽

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.

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