Establishment of Offline 2D-LC-MS System and Its Application to Human Liver Proteomics

Abstract

Abstract: A lab-made off line 2D-LC-MS system was developed for protein analysis. In case the off line 2D-LC was utilized for peptide separation, and the working conditions for strong cation exchange(SCX) chromatography and the succeeding reversed phase(RP) chromatography can be individually optimized, contrasted to the on-line system. The experimental results show that the mass fractions of acetonitrile for the SCX separation can be increased form 5% as references reported up to 20% so that the separation of peptides has been substantially improved. The off line system has been applied to the analysis of proteins extracted from the cancer tissue of nude mice inoculated by a human-hepatocellular-carcinoma cell-line D20. Some extreme basic proteins(pI>10) and larger proteins(M_w>200 000), which are difficult to be isolated by the traditional 2D-PAGE method, have been identified in D20 sample.

Key words: Multidimensional-liquid chromatography-mass spectrometry, Offline 2D-LC-MS, Water-soluble proteins

CLC Number:

O657

TrendMD:

WANG Jing, XU Xue-Jiao, YU Yan-Ling, LIU Yin-Kun, YANG Peng-Yuan, . Establishment of Offline 2D-LC-MS System and Its Application to Human Liver Proteomics[J]. Chem. J. Chinese Universities, 2004, 25(12): 2247.

[1]	SUN Yang, ZHANG Lei, XU Fei, GONG Bo-Lin. Preparation of Chiral Ligand Exchange Chromatographic Stationary Phase by Atom Transfer Radical Polymerization for Separation of Racemates [J]. Chem. J. Chinese Universities, 2012, 33(12): 2644.
[2]	ZHU Ying, LIU Pei, YANG Xiao-Hai, HE Lei-Liang, LI Qing-Zhao, WANG Qing, WANG Ke-Min, HUANG Jin, LIU Jian-Bo. Single-labeled“Turn on” Fluorescent Oligonucleotide Probe for Iodide Detection Based on T-Hg²⁺-T and Inherent Quenching Ability of G-quadruplex [J]. Chem. J. Chinese Universities, 2012, 33(12): 2651.
[3]	TIAN Qing-Qing, ZENG Wei-Xiu, ZHAO Xin, HOU Yu-Lan, CHEN Bo, YAO Shou-Zhuo. Solvent Induced Phase Separation Extraction-Spectrophotometric Method for Determination of Angiotensin-converting Enzyme Activity [J]. Chem. J. Chinese Universities, 2012, 33(11): 2417.
[4]	HE Wei-Na, TU Xi-Juan, GUO Xiang-Qun. Spin-Labeled BSA-modified Near-infrared Fluorescent Gold Nanoparticles [J]. Chem. J. Chinese Universities, 2012, 33(11): 2411.
[5]	YANG Jun, QI Li, MA Hui-Min, CHEN Yi. Fabrication of Nylon Membrane-based Microfluidic Chips and Its Application in Color Sensing of Glucose [J]. Chem. J. Chinese Universities, 2012, 33(11): 2405.
[6]	CAO Xia, ZENG Xi, MU Lan, CHEN Yi, WANG Rui-Xiao, ZHANG Yun-Qian, ZHANG Jian-Xin, WEI Gang. Synthesis, Characterization and Aggregation Induced Enhanced Emission of New Phenothiazine Hydrazone [J]. Chem. J. Chinese Universities, 2012, 33(10): 2184.
[7]	WANG Qing, YANG Li-Yan, LIU Jian-Bo, YANG Xiao-Hai, WANG Ke-Min, JI Hai-Ning, LIU Yan. Dynamic Light Scattering for Fluoride Ions Detection [J]. Chem. J. Chinese Universities, 2012, 33(10): 2195.
[8]	LUO Ya-Hui, LI Gang, CHEN Qiang, ZHAO Jian-Long. Microfluidic Device for Automated High-Throughput Protein Crystallization Screening by Osmotic Dewatering [J]. Chem. J. Chinese Universities, 2012, 33(10): 2178.
[9]	HAO Li, CHENG He-Yong, LIU Jin-Hua, YIN Xue-Feng. Direct Determination of Lead and Cadmium in Distilled Spirits by Inductively Coupled Plasma Mass Spectrometry Coupled with Chip-based Microflow Injection System [J]. Chem. J. Chinese Universities, 2012, 33(09): 1957.
[10]	LIU Yue-Qin, PI Zi-Feng, SONG Feng-Rui, LIU Zhi-Qiang, LIU Zhong-Ying. Effect of Acarbose on Urine Metabolic Profiling of Type Ⅱ Diabete Rats [J]. Chem. J. Chinese Universities, 2012, 33(09): 1932.
[11]	FENG Ke-Jun, YAO Yan-Ling, SHEN Guo-Li, YU Ru-Qin. Single-walled Carbon Nanotube/DNAzyme Complex-based Elctrochemical Biosensors [J]. Chem. J. Chinese Universities, 2012, 33(08): 1676.
[12]	ZHAO Qiang, SUN Xiao-Hong, LU Ying, PAN Ying-Jie, ZHAO Yong. Application of Metabolic Profiling for Distinguishing Different Pathogenic Vibrio Parahaemolyticus [J]. Chem. J. Chinese Universities, 2012, 33(08): 1686.
[13]	WANG Lin, TAN Xue-Cai, ZHAO Dan-Dan, LIU Li, LEI Fu-Hou, HUANG Zai-Yin, GONG Qi. Electrochemical Sensor for Caffeine Detection Based on Molecularly Imprinted Polymer with Maleic Rosin Acrylic Acid Glycol Ester as Cross-linker [J]. Chem. J. Chinese Universities, 2012, 33(08): 1708.
[14]	LIU Xue-Ping, OUYANG Xiang-Yuan, WU Hui-Wang, SHEN Guo-Li, YU Ru-Qin. Novel Fluorescent Technique for Detecting Adenosine Based on DNA Strand Displacement Induced by Target-aptamer Complex and Metal Deposition Catalyzed by Gold Nanoparticles [J]. Chem. J. Chinese Universities, 2012, 33(08): 1692.
[15]	SUN Shu-Guo, MA Mei-Hu, QIN Fang, XU Ya-Ru, YU Wen-Zhen, LUO Zhang. Covalent Immobilization of Avidin onto Magnetic Nanoparticles and Application in Rapid Determination of Protein A [J]. Chem. J. Chinese Universities, 2012, 33(07): 1426.

Establishment of Offline 2D-LC-MS System and Its Application to Human Liver Proteomics

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments