Separation of proteins by aqueous two-phase extraction system combined with liquid chromatography

Xin Ying Zhao; Feng Qu; Min Dong; Fan Chen; Ai Qin Luo; Jing Hua Zhang

doi:10.1016/S1872-2040(11)60520-3

Separation of proteins by aqueous two-phase extraction system combined with liquid chromatography

Xin Ying Zhao
, Feng Qu^*
, Min Dong
, Fan Chen
, Ai Qin Luo
, Jing Hua Zhang

^*Corresponding author for this work

School of Life Science

Research output: Contribution to journal › Article › peer-review

11 Citations (Scopus)

Abstract

A method was developed for the enrichment and separation of proteins by an aqueous two-phase extraction system (ATPS) of PEG/(NH₄) ₂SO₄ combined with high-performance liquid chromatography (HPLC). The effects of the type and the concentration of inorganic salts, PEG molecular weight, pH value, temperature and NaCl on the formation of two phases and partition of cytochrome c, lysozyme, trypsin, bovine serum albumin and myoglobin were investigated, respectively. The results showed that five proteins were enriched mainly in bottom phase in 15% PEG-4000/10% (NH₄) ₂SO₄ at room temperature and pH from 3.5 to 9.0. High-abundance proteins in serum were also distributed in two phases and the bottom phase had larger amount under the same conditions, which could be extracted and separated by optimized ATPS. The proteins were collected on the basis of their retention time in HPLC, and could be removed from serum samples.

Original language	English
Pages (from-to)	38-42
Number of pages	5
Journal	Chinese Journal of Analytical Chemistry
Volume	40
Issue number	1
DOIs	https://doi.org/10.1016/S1872-2040(11)60520-3
Publication status	Published - Jan 2012

Keywords

Aqueous two-phase extraction
Proteins separation
Serum

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10.1016/S1872-2040(11)60520-3

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title = "Separation of proteins by aqueous two-phase extraction system combined with liquid chromatography",

abstract = "A method was developed for the enrichment and separation of proteins by an aqueous two-phase extraction system (ATPS) of PEG/(NH4) 2SO4 combined with high-performance liquid chromatography (HPLC). The effects of the type and the concentration of inorganic salts, PEG molecular weight, pH value, temperature and NaCl on the formation of two phases and partition of cytochrome c, lysozyme, trypsin, bovine serum albumin and myoglobin were investigated, respectively. The results showed that five proteins were enriched mainly in bottom phase in 15\% PEG-4000/10\% (NH4) 2SO4 at room temperature and pH from 3.5 to 9.0. High-abundance proteins in serum were also distributed in two phases and the bottom phase had larger amount under the same conditions, which could be extracted and separated by optimized ATPS. The proteins were collected on the basis of their retention time in HPLC, and could be removed from serum samples.",

keywords = "Aqueous two-phase extraction, Proteins separation, Serum",

author = "Zhao, \{Xin Ying\} and Feng Qu and Min Dong and Fan Chen and Luo, \{Ai Qin\} and Zhang, \{Jing Hua\}",

year = "2012",

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doi = "10.1016/S1872-2040(11)60520-3",

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TY - JOUR

T1 - Separation of proteins by aqueous two-phase extraction system combined with liquid chromatography

AU - Zhao, Xin Ying

AU - Qu, Feng

AU - Dong, Min

AU - Chen, Fan

AU - Luo, Ai Qin

AU - Zhang, Jing Hua

PY - 2012/1

Y1 - 2012/1

N2 - A method was developed for the enrichment and separation of proteins by an aqueous two-phase extraction system (ATPS) of PEG/(NH4) 2SO4 combined with high-performance liquid chromatography (HPLC). The effects of the type and the concentration of inorganic salts, PEG molecular weight, pH value, temperature and NaCl on the formation of two phases and partition of cytochrome c, lysozyme, trypsin, bovine serum albumin and myoglobin were investigated, respectively. The results showed that five proteins were enriched mainly in bottom phase in 15% PEG-4000/10% (NH4) 2SO4 at room temperature and pH from 3.5 to 9.0. High-abundance proteins in serum were also distributed in two phases and the bottom phase had larger amount under the same conditions, which could be extracted and separated by optimized ATPS. The proteins were collected on the basis of their retention time in HPLC, and could be removed from serum samples.

AB - A method was developed for the enrichment and separation of proteins by an aqueous two-phase extraction system (ATPS) of PEG/(NH4) 2SO4 combined with high-performance liquid chromatography (HPLC). The effects of the type and the concentration of inorganic salts, PEG molecular weight, pH value, temperature and NaCl on the formation of two phases and partition of cytochrome c, lysozyme, trypsin, bovine serum albumin and myoglobin were investigated, respectively. The results showed that five proteins were enriched mainly in bottom phase in 15% PEG-4000/10% (NH4) 2SO4 at room temperature and pH from 3.5 to 9.0. High-abundance proteins in serum were also distributed in two phases and the bottom phase had larger amount under the same conditions, which could be extracted and separated by optimized ATPS. The proteins were collected on the basis of their retention time in HPLC, and could be removed from serum samples.

KW - Aqueous two-phase extraction

KW - Proteins separation

KW - Serum

UR - https://www.scopus.com/pages/publications/84863061560

U2 - 10.1016/S1872-2040(11)60520-3

DO - 10.1016/S1872-2040(11)60520-3

M3 - Article

AN - SCOPUS:84863061560

SN - 0253-3820

VL - 40

SP - 38

EP - 42

JO - Chinese Journal of Analytical Chemistry

JF - Chinese Journal of Analytical Chemistry

IS - 1

ER -

Separation of proteins by aqueous two-phase extraction system combined with liquid chromatography

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Keywords

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