Development and evaluation of a double antibody sandwich ELISA for the detection of human sDC-SIGN.
J Immunol Methods
; 436: 16-21, 2016 09.
Article
em En
| MEDLINE
| ID: mdl-27262264
ABSTRACT
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.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Células Dendríticas
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Ensaio de Imunoadsorção Enzimática
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Moléculas de Adesão Celular
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Receptores de Superfície Celular
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Hepatite B Crônica
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Lectinas Tipo C
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Neoplasias
Tipo de estudo:
Diagnostic_studies
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Evaluation_studies
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Observational_studies
Limite:
Animals
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Female
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Humans
Idioma:
En
Revista:
J Immunol Methods
Ano de publicação:
2016
Tipo de documento:
Article
País de afiliação:
China