Purification, identification and profiling of serum amyloid A proteins from sera of advanced-stage cancer patients

Jing Li; Zhensheng Xie; Linan Shi; Zhiqiang Zhao; Junjie Hou; Xiulan Chen; Ziyou Cui; Peng Xue; Tanxi Cai; Peng Wu; Sutang Guo; Fuquan Yang

doi:10.1016/j.jchromb.2012.01.002

Purification, identification and profiling of serum amyloid A proteins from sera of advanced-stage cancer patients

Jing Li, Zhensheng Xie, Linan Shi, Zhiqiang Zhao, Junjie Hou, Xiulan Chen, Ziyou Cui, Peng Xue, Tanxi Cai, Peng Wu, Sutang Guo, Fuquan Yang

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

16 Scopus citations

Abstract

Surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) is a powerful tool for screening potential biomarkers of various pathological conditions. However, low resolution and mass accuracy of SELDI-TOF-MS remain a major obstacle for determination of biological identities of potential protein biomarkers. We report here a refined workflow that combines ZipTip desalting, acetonitrile precipitation, high-performance liquid chromatography (HPLC) separation and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) analysis for the profiling, purification and identification of the targeted serum proteins found by SELDI-TOF-MS. By using this workflow, we purified ten targeted proteins from the sera of patients with various types of advanced stage (stage III-IV) cancers. These proteins were identified as isoforms of the human serum amyloid protein A (SAA) family with or without truncations at their N-terminals. This was confirmed by Western blot analysis. Different SAA expression patterns were observed by MALDI-TOF-MS profiling. SAA has long been reported as a biomarker for various cancer types such as lung cancer, ovarian cancer, and breast cancer. However, in this study we found increased SAA expression in the sera of advanced-stage cancer patients with different cancer types. Our results suggest that maybe SAA should not be used alone as a biomarker for any specific cancer type.

Original language	English (US)
Pages (from-to)	3-9
Number of pages	7
Journal	Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume	889-890
DOIs	https://doi.org/10.1016/j.jchromb.2012.01.002
State	Published - Mar 15 2012
Externally published	Yes

Bibliographical note

Funding Information:
This research was supported by the National Basic Research Program of China (973) (grant nos. 2010CB833703 , 2012CB966803 , 2011CB915501 ) and the National Natural Science Foundation of China (grant no. 90919047 ).

Keywords

MALDI-TOF-MS
Purification
SAA
SELDI-TOF-MS
Targeted protein

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Publisher link

10.1016/j.jchromb.2012.01.002

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Li, J., Xie, Z., Shi, L., Zhao, Z., Hou, J., Chen, X., Cui, Z., Xue, P., Cai, T., Wu, P., Guo, S., & Yang, F. (2012). Purification, identification and profiling of serum amyloid A proteins from sera of advanced-stage cancer patients. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 889-890, 3-9. https://doi.org/10.1016/j.jchromb.2012.01.002

Li, J, Xie, Z, Shi, L, Zhao, Z, Hou, J, Chen, X, Cui, Z, Xue, P, Cai, T, Wu, P, Guo, S & Yang, F 2012, 'Purification, identification and profiling of serum amyloid A proteins from sera of advanced-stage cancer patients', Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, vol. 889-890, pp. 3-9. https://doi.org/10.1016/j.jchromb.2012.01.002

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abstract = "Surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) is a powerful tool for screening potential biomarkers of various pathological conditions. However, low resolution and mass accuracy of SELDI-TOF-MS remain a major obstacle for determination of biological identities of potential protein biomarkers. We report here a refined workflow that combines ZipTip desalting, acetonitrile precipitation, high-performance liquid chromatography (HPLC) separation and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) analysis for the profiling, purification and identification of the targeted serum proteins found by SELDI-TOF-MS. By using this workflow, we purified ten targeted proteins from the sera of patients with various types of advanced stage (stage III-IV) cancers. These proteins were identified as isoforms of the human serum amyloid protein A (SAA) family with or without truncations at their N-terminals. This was confirmed by Western blot analysis. Different SAA expression patterns were observed by MALDI-TOF-MS profiling. SAA has long been reported as a biomarker for various cancer types such as lung cancer, ovarian cancer, and breast cancer. However, in this study we found increased SAA expression in the sera of advanced-stage cancer patients with different cancer types. Our results suggest that maybe SAA should not be used alone as a biomarker for any specific cancer type.",

keywords = "MALDI-TOF-MS, Purification, SAA, SELDI-TOF-MS, Targeted protein",

author = "Jing Li and Zhensheng Xie and Linan Shi and Zhiqiang Zhao and Junjie Hou and Xiulan Chen and Ziyou Cui and Peng Xue and Tanxi Cai and Peng Wu and Sutang Guo and Fuquan Yang",

note = "Funding Information: This research was supported by the National Basic Research Program of China (973) (grant nos. 2010CB833703 , 2012CB966803 , 2011CB915501 ) and the National Natural Science Foundation of China (grant no. 90919047 ). ",

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doi = "10.1016/j.jchromb.2012.01.002",

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T1 - Purification, identification and profiling of serum amyloid A proteins from sera of advanced-stage cancer patients

AU - Li, Jing

AU - Xie, Zhensheng

AU - Shi, Linan

AU - Zhao, Zhiqiang

AU - Hou, Junjie

AU - Chen, Xiulan

AU - Cui, Ziyou

AU - Xue, Peng

AU - Cai, Tanxi

AU - Wu, Peng

AU - Guo, Sutang

AU - Yang, Fuquan

N1 - Funding Information: This research was supported by the National Basic Research Program of China (973) (grant nos. 2010CB833703 , 2012CB966803 , 2011CB915501 ) and the National Natural Science Foundation of China (grant no. 90919047 ).

PY - 2012/3/15

Y1 - 2012/3/15

N2 - Surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) is a powerful tool for screening potential biomarkers of various pathological conditions. However, low resolution and mass accuracy of SELDI-TOF-MS remain a major obstacle for determination of biological identities of potential protein biomarkers. We report here a refined workflow that combines ZipTip desalting, acetonitrile precipitation, high-performance liquid chromatography (HPLC) separation and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) analysis for the profiling, purification and identification of the targeted serum proteins found by SELDI-TOF-MS. By using this workflow, we purified ten targeted proteins from the sera of patients with various types of advanced stage (stage III-IV) cancers. These proteins were identified as isoforms of the human serum amyloid protein A (SAA) family with or without truncations at their N-terminals. This was confirmed by Western blot analysis. Different SAA expression patterns were observed by MALDI-TOF-MS profiling. SAA has long been reported as a biomarker for various cancer types such as lung cancer, ovarian cancer, and breast cancer. However, in this study we found increased SAA expression in the sera of advanced-stage cancer patients with different cancer types. Our results suggest that maybe SAA should not be used alone as a biomarker for any specific cancer type.

AB - Surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) is a powerful tool for screening potential biomarkers of various pathological conditions. However, low resolution and mass accuracy of SELDI-TOF-MS remain a major obstacle for determination of biological identities of potential protein biomarkers. We report here a refined workflow that combines ZipTip desalting, acetonitrile precipitation, high-performance liquid chromatography (HPLC) separation and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) analysis for the profiling, purification and identification of the targeted serum proteins found by SELDI-TOF-MS. By using this workflow, we purified ten targeted proteins from the sera of patients with various types of advanced stage (stage III-IV) cancers. These proteins were identified as isoforms of the human serum amyloid protein A (SAA) family with or without truncations at their N-terminals. This was confirmed by Western blot analysis. Different SAA expression patterns were observed by MALDI-TOF-MS profiling. SAA has long been reported as a biomarker for various cancer types such as lung cancer, ovarian cancer, and breast cancer. However, in this study we found increased SAA expression in the sera of advanced-stage cancer patients with different cancer types. Our results suggest that maybe SAA should not be used alone as a biomarker for any specific cancer type.

KW - MALDI-TOF-MS

KW - Purification

KW - SAA

KW - SELDI-TOF-MS

KW - Targeted protein

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SN - 1570-0232

VL - 889-890

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JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

ER -

Purification, identification and profiling of serum amyloid A proteins from sera of advanced-stage cancer patients

Abstract

Bibliographical note

Keywords

UN SDGs

Publisher link

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