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

15 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)

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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.",

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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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AU - Li, Jing

AU - Xie, Zhensheng

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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 ).

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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.

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

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ER -

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

Abstract

Bibliographical note

Keywords

UN SDGs

Access

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