title: Detection of Human Epididymis Protein 4 (HE4) in Human Serum Samples Using a Specific Monoclonal Antibody-Based Sandwich Enzyme-Linked Immunosorbent Assay (ELISA).
abstract: BACKGROUND
  The human epididymis protein 4 (HE4) may have high specificity in the detection of malignant diseases, making the development of an immunoassay for HE4 essential.
  
  
  METHODS
  In our study, a fusion gene was constructed encoded with the HE4 protein. This protein was then produced in the bacterial cells (Escherichia coli) and used to immunize mice in order to eventually generate hybridomas specific to HE4. The hybridoma supernatants were then screened, and four positive anti-HE4 cell lines were selected. These cell lines produce monoclonal antibodies against HE4 epitopes, as demonstrated in the Western blot as well as by direct enzyme-linked immunosorbent assay (ELISA). Using the developed antibodies, we successfully identified several good antibody pairs from the hybridomas, which allowed for the development of a sandwich ELISA to measure HE4 levels. By using the HE4 ELISA, we measured HE4 levels of 60 clinical human serum samples.
  
  
  RESULTS
  Compared with the Food and Drug Administration (FDA) approved kit (Roche), our results showed a strong positive correlation to those of the FDA-approved kit.
  
  
  CONCLUSIONS
  In summary, highly sensitive antibody pairs were screened against HE4, and a sandwich ELISA was developed as an accurate analytical tool for the detection of HE4 in human serum, which could be especially valuable for diagnosing ovarian carcinomas.
authors.affiliation: Department of Clinical Laboratory, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Raybiotech, Inc, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Lv
authors.affiliation: Medical Research Center, the Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Shao
authors.affiliation: Department of Clinical Laboratory, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Clinical Laboratory, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Luo
authors.affiliation: Raybiotech, Inc, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Yuming
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Raybiotech, Inc, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Raybiotech, Inc, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Wenji
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Raybiotech, Inc, Guangzhou, Guangdong Province, People's Republic of China.|RayBiotech, Inc, Norcross, Georgia.|South China Biochip Research Center, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Shuhong
authors.initials: S
authors.lastname: Luo
authors.affiliation: Raybiotech, Inc, Guangzhou, Guangdong Province, People's Republic of China.|RayBiotech, Inc, Norcross, Georgia.|South China Biochip Research Center, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Jianmin
authors.initials: J
authors.lastname: Fang
authors.affiliation: Department of Clinical Laboratory, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Clinical Laboratory, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, Guangdong Province, People's Republic of China. 1725012289@qq.com.
authors.forename: Chaohui
authors.initials: C
authors.lastname: Duan
authors.affiliation: Raybiotech, Inc, Guangzhou, Guangdong Province, People's Republic of China. rhuang@raybiotech.com.|RayBiotech, Inc, Norcross, Georgia. rhuang@raybiotech.com.|South China Biochip Research Center, Guangzhou, Guangdong Province, People's Republic of China. rhuang@raybiotech.com.
authors.forename: Ruopan
authors.initials: R
authors.lastname: Huang
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism*; D000918:Antibody Specificity; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D051379:Mice; D008875:Middle Aged; D010051:Ovarian Neoplasms / Q000097:blood*; D011506:Proteins / Q000276:immunology* / Q000378:metabolism*; D012680:Sensitivity and Specificity; D000081102:WAP Four-Disulfide Core Domain Protein 2; D055815:Young Adult
chemical_list: D000911:Antibodies, Monoclonal; D011506:Proteins; D000081102:WAP Four-Disulfide Core Domain Protein 2; C569698:WFDC2 protein, human
keywords: HE4; immunoassay; monoclonal antibody; ovarian cancer; sandwich ELISA