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

Research output: Contribution to journalArticle

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 languageEnglish (US)
Pages (from-to)61-67
Number of pages7
JournalJournal of Immunological Methods
Volume355
Issue number1-2
DOIs
StatePublished - Apr 2010

Keywords

  • Hepatoma-derived growth factor
  • Immunohistochemistry
  • MAb
  • NSCLC
  • Sandwich ELISA

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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