Development and evaluation of a double antibody sandwich ELISA for the detection of human sDC-SIGN.
sDC-SIGN is the soluble form of dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN, CD209), which is a molecule involved with pathogen recognition and immune regulation. However, there is no commercially available ELISA kit for detecting human sDC-SIGN, and the normal range of this molecule is unknown. Here, we describe an ELISA for detecting human sDC-SIGN with high specificity. First, sDC-SIGN protein was expressed and purified. Monoclonal and polyclonal antibodies were then raised against the purified protein and subsequently characterized. A sandwich ELISA was developed using polyclonal antibodies specific for sDC-SIGN for capture and a biotin-labeled monoclonal antibody specific for sDC-SIGN for detection of protein. This method has sensitivity up to 0.2 ng/ml. Using this ELISA, we found that the concentration of sDC-SIGN in sera of healthy volunteers ranges from 0-319 ng/ml with a mean concentration of 27.14 ng/ml. Interestingly, the concentration of sDC-SIGN in sera from patients with cancer or chronic hepatitis B virus (CHB) infection was lower than that of health controls. The mean concentrations of sDC-SIGN in cancer patients and chronic hepatitis B virus infection patients were 3.2 ng/ml and 3.8 ng/ml, respectively. We developed a sandwich ELISA for detecting human sDC-SIGN and demonstrated its use by assessing sera concentrations of sDC-SIGN in patients with cancer and chronic CHB infection compared to that of healthy controls.
Department of Immunology, Southern Medical University, Guangzhou, China.
Shang-Liang
SL
Chen
Department of Immunology, Southern Medical University, Guangzhou, China.
Yan-Li
YL
Li
Department of Immunology, Southern Medical University, Guangzhou, China.
Yuan
Y
Tang
Clinical Laboratory, Second People's Hospital of Foshan, Foshan, China.
Zhi-Cheng
ZC
Chen
Department of Immunology, Southern Medical University, Guangzhou, China.
Jing
J
Zhou
Department of Immunology, Southern Medical University, Guangzhou, China.
Jia
J
Zhou
Department of Immunology, Southern Medical University, Guangzhou, China.
Xiao
X
Lu
Department of Immunology, Southern Medical University, Guangzhou, China.
Na
N
Zhao
Department of Immunology, Southern Medical University, Guangzhou, China; Guangdong Province Key Laboratory of Proteomics, Southern Medical University, Guangzhou, China. Electronic address: zhlchen@smu.edu.cn.
Zheng-Liang
ZL
Chen
Department of Immunology, Southern Medical University, Guangzhou, China. Electronic address: zdaming@smu.edu.cn.
Daming
D
Zuo
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016022:Case-Control Studies; D015815:Cell Adhesion Molecules / Q000097:blood / Q000302:isolation & purification*; D003713:Dendritic Cells / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D019694:Hepatitis B, Chronic / Q000097:blood* / Q000175:diagnosis; D006801:Humans; D037181:Lectins, C-Type / Q000097:blood / Q000302:isolation & purification*; D013058:Mass Spectrometry; D051379:Mice; D008807:Mice, Inbred BALB C; D009369:Neoplasms / Q000097:blood* / Q000175:diagnosis; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000097:blood / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
D000911:Antibodies, Monoclonal; D015815:Cell Adhesion Molecules; D037181:Lectins, C-Type; D011956:Receptors, Cell Surface; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
CD209; chronic hepatitis B virus infection; sDC-SIGN; sandwich ELISA