Development and clinical evaluation of a multi-purpose mAb and a sandwich ELISA test for hepatoma-derived growth factor in lung cancer patients

Aixia Zhang; Weiguo Long; Ze Guo; Genyan Liu; Zhibin Hu; Yan Huang; Yunqian Li; Tessa M. Grabinski; Jun Yang; Ping X. Zhao; Allen D. Everett; Yan Zhang; Brian B. Cao

doi:10.1016/j.jim.2010.02.011

Development and clinical evaluation of a multi-purpose mAb and a sandwich ELISA test for hepatoma-derived growth factor in lung cancer patients

Aixia Zhang, Weiguo Long, Ze Guo, Genyan Liu, Zhibin Hu, Yan Huang, Yunqian Li, Tessa M. Grabinski, Jun Yang, Ping X. Zhao, Allen D. Everett, Yan Zhang, Brian B. Cao

School of Medicine

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

9 Scopus citations

Abstract

Hepatoma-derived growth factor (HDGF) is closely related to aggressive tumor behavior and could be a broader biomarker for cancer prognosis and diagnosis. The goal of this study is to develop a sandwich ELISA system to test if HDGF can be detected in serum samples. We produced an anti-HDGF monoclonal antibody designated 2F12 using recombinant human HDGF protein. The specificity of 2F12 mAb was characterized by western blotting, immunohistochemistry (IHC), immunofluorescent staining (IF) and immunoprecipitation (IP). The results showed that 2F12 recognized HDGF in both native and denatured form, and can be used for multiple purposes. We have found that HDGF is also expressed in several cancers unreported previously by IHC staining on tumor cell array sections. In addition, we have developed a sandwich ELISA assay using mAb 2F12 and rabbit anti-HDGF polyclonal antibody, and validated the assay using normal serum and non-small cell lung cancer (NSCLC) serum samples. The sensitivity of this assay is 0.5. ng/ml and the linear range is 0.5-32 ng/ml. The HDGF average level in serum samples from lung cancer patients is significantly elevated relative to that from healthy controls, 9.43 ± 6.13 ng/ml versus 4.36 ± 2.50 ng/ml (p=1.12E-10).

Original language	English (US)
Pages (from-to)	61-67
Number of pages	7
Journal	Journal of Immunological Methods
Volume	355
Issue number	1-2
DOIs	https://doi.org/10.1016/j.jim.2010.02.011
State	Published - Apr 2010

Keywords

Hepatoma-derived growth factor
Immunohistochemistry
MAb
NSCLC
Sandwich ELISA

ASJC Scopus subject areas

Immunology and Allergy
Immunology

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10.1016/j.jim.2010.02.011

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Zhang, A., Long, W., Guo, Z., Liu, G., Hu, Z., Huang, Y., Li, Y., Grabinski, T. M., Yang, J., Zhao, P. X., Everett, A. D., Zhang, Y., & Cao, B. B. (2010). Development and clinical evaluation of a multi-purpose mAb and a sandwich ELISA test for hepatoma-derived growth factor in lung cancer patients. Journal of Immunological Methods, 355(1-2), 61-67. https://doi.org/10.1016/j.jim.2010.02.011

Zhang, A, Long, W, Guo, Z, Liu, G, Hu, Z, Huang, Y, Li, Y, Grabinski, TM, Yang, J, Zhao, PX, Everett, AD, Zhang, Y & Cao, BB 2010, 'Development and clinical evaluation of a multi-purpose mAb and a sandwich ELISA test for hepatoma-derived growth factor in lung cancer patients', Journal of Immunological Methods, vol. 355, no. 1-2, pp. 61-67. https://doi.org/10.1016/j.jim.2010.02.011

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title = "Development and clinical evaluation of a multi-purpose mAb and a sandwich ELISA test for hepatoma-derived growth factor in lung cancer patients",

abstract = "Hepatoma-derived growth factor (HDGF) is closely related to aggressive tumor behavior and could be a broader biomarker for cancer prognosis and diagnosis. The goal of this study is to develop a sandwich ELISA system to test if HDGF can be detected in serum samples. We produced an anti-HDGF monoclonal antibody designated 2F12 using recombinant human HDGF protein. The specificity of 2F12 mAb was characterized by western blotting, immunohistochemistry (IHC), immunofluorescent staining (IF) and immunoprecipitation (IP). The results showed that 2F12 recognized HDGF in both native and denatured form, and can be used for multiple purposes. We have found that HDGF is also expressed in several cancers unreported previously by IHC staining on tumor cell array sections. In addition, we have developed a sandwich ELISA assay using mAb 2F12 and rabbit anti-HDGF polyclonal antibody, and validated the assay using normal serum and non-small cell lung cancer (NSCLC) serum samples. The sensitivity of this assay is 0.5. ng/ml and the linear range is 0.5-32 ng/ml. The HDGF average level in serum samples from lung cancer patients is significantly elevated relative to that from healthy controls, 9.43 ± 6.13 ng/ml versus 4.36 ± 2.50 ng/ml (p=1.12E-10).",

keywords = "Hepatoma-derived growth factor, Immunohistochemistry, MAb, NSCLC, Sandwich ELISA",

author = "Aixia Zhang and Weiguo Long and Ze Guo and Genyan Liu and Zhibin Hu and Yan Huang and Yunqian Li and Grabinski, {Tessa M.} and Jun Yang and Zhao, {Ping X.} and Everett, {Allen D.} and Yan Zhang and Cao, {Brian B.}",

note = "Funding Information: We thank David Nadziejka for editorial reading of the manuscript. This work was partially supported by the Jay and Betty Van Andel Foundation .",

year = "2010",

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doi = "10.1016/j.jim.2010.02.011",

language = "English (US)",

volume = "355",

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T1 - Development and clinical evaluation of a multi-purpose mAb and a sandwich ELISA test for hepatoma-derived growth factor in lung cancer patients

AU - Zhang, Aixia

AU - Long, Weiguo

AU - Guo, Ze

AU - Liu, Genyan

AU - Hu, Zhibin

AU - Huang, Yan

AU - Li, Yunqian

AU - Grabinski, Tessa M.

AU - Yang, Jun

AU - Zhao, Ping X.

AU - Everett, Allen D.

AU - Zhang, Yan

AU - Cao, Brian B.

N1 - Funding Information: We thank David Nadziejka for editorial reading of the manuscript. This work was partially supported by the Jay and Betty Van Andel Foundation .

PY - 2010/4

Y1 - 2010/4

N2 - Hepatoma-derived growth factor (HDGF) is closely related to aggressive tumor behavior and could be a broader biomarker for cancer prognosis and diagnosis. The goal of this study is to develop a sandwich ELISA system to test if HDGF can be detected in serum samples. We produced an anti-HDGF monoclonal antibody designated 2F12 using recombinant human HDGF protein. The specificity of 2F12 mAb was characterized by western blotting, immunohistochemistry (IHC), immunofluorescent staining (IF) and immunoprecipitation (IP). The results showed that 2F12 recognized HDGF in both native and denatured form, and can be used for multiple purposes. We have found that HDGF is also expressed in several cancers unreported previously by IHC staining on tumor cell array sections. In addition, we have developed a sandwich ELISA assay using mAb 2F12 and rabbit anti-HDGF polyclonal antibody, and validated the assay using normal serum and non-small cell lung cancer (NSCLC) serum samples. The sensitivity of this assay is 0.5. ng/ml and the linear range is 0.5-32 ng/ml. The HDGF average level in serum samples from lung cancer patients is significantly elevated relative to that from healthy controls, 9.43 ± 6.13 ng/ml versus 4.36 ± 2.50 ng/ml (p=1.12E-10).

AB - Hepatoma-derived growth factor (HDGF) is closely related to aggressive tumor behavior and could be a broader biomarker for cancer prognosis and diagnosis. The goal of this study is to develop a sandwich ELISA system to test if HDGF can be detected in serum samples. We produced an anti-HDGF monoclonal antibody designated 2F12 using recombinant human HDGF protein. The specificity of 2F12 mAb was characterized by western blotting, immunohistochemistry (IHC), immunofluorescent staining (IF) and immunoprecipitation (IP). The results showed that 2F12 recognized HDGF in both native and denatured form, and can be used for multiple purposes. We have found that HDGF is also expressed in several cancers unreported previously by IHC staining on tumor cell array sections. In addition, we have developed a sandwich ELISA assay using mAb 2F12 and rabbit anti-HDGF polyclonal antibody, and validated the assay using normal serum and non-small cell lung cancer (NSCLC) serum samples. The sensitivity of this assay is 0.5. ng/ml and the linear range is 0.5-32 ng/ml. The HDGF average level in serum samples from lung cancer patients is significantly elevated relative to that from healthy controls, 9.43 ± 6.13 ng/ml versus 4.36 ± 2.50 ng/ml (p=1.12E-10).

KW - Hepatoma-derived growth factor

KW - Immunohistochemistry

KW - MAb

KW - NSCLC

KW - Sandwich ELISA

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Development and clinical evaluation of a multi-purpose mAb and a sandwich ELISA test for hepatoma-derived growth factor in lung cancer patients

Abstract

Keywords

ASJC Scopus subject areas

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