Development of a phage display-mediated immunoassay for the detection of vascular endothelial growth factor

Autor:	S. Shirin Shahangian, Reza H. Sajedi, Sadegh Hasannia, Zahra Rezaei
Rok vydání:	2020
Předmět:	Vascular Endothelial Growth Factor A Phage display medicine.drug_class Angiogenesis Enzyme-Linked Immunosorbent Assay 02 engineering and technology Monoclonal antibody Polymerase Chain Reaction 01 natural sciences Biochemistry Analytical Chemistry chemistry.chemical_compound Limit of Detection medicine Humans Detection limit biology medicine.diagnostic_test Chemistry 010401 analytical chemistry 021001 nanoscience & nanotechnology Molecular biology 0104 chemical sciences Vascular endothelial growth factor Single-domain antibody Immunoassay biology.protein Antibody Cell Surface Display Techniques 0210 nano-technology Antibodies Immobilized
Zdroj:	Analytical and Bioanalytical Chemistry. 412:7639-7648
ISSN:	1618-2650 1618-2642
DOI:	10.1007/s00216-020-02901-4
Popis:	Because of the critical role of vascular endothelial growth factor (VEGF) in angiogenesis and its significantly increased serum levels in early stages of cancer, VEGF is considered an important prognostic biomarker in different cancers. Herein, the amplification power of PCR combined with phage displaying anti-VEGF VHH, a sensitive real-time immunoassay, was precisely designed based on phage display-mediated immuno-PCR (PD-IPCR) for the detection of VEGF. This system benefits from strong and specific binding of antigen and antibody in a sandwich immunosorbent assay platform using avastin (anti-VEGF monoclonal antibody) as the capture antibody. The anti-VEGF phage particles were used as both anti-VEGF agent and DNA template in the PD-IPCR. Anti-VEGF phage ELISA showed a linear range of 3–250 ng/ml and a limit of detection (LOD) of 1.1 ng/ml. Using the PD-IPCR method, the linear range of VEGF detection was found to be 0.06–700 ng/ml, with a detection limit of 3 pg/ml. The recovery rate in serum ranged from 83% to 99%, with a relative standard deviation of 1.2–4.9%. These values indicate that the method has good sensitivity for use in clinical analysis. The proposed method was successfully applied to the clinical determination of VEGF in human serum samples, and the results showed excellent correlation with conventional ELISA (R2 = 0.995). The novel immunoassay provides a specific and sensitive immunoassay protocol for VEGF detection at very low levels.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0c703869df2c6c48c7e8e96f05e50b8d https://doi.org/10.1007/s00216-020-02901-4 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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