Green fluorescent-conjugated anti-CEA single chain antibody for the detection of CEA-positive cancer cells

Autor: Maryam Salavatifar, Zahra Moghaddassi Jahromi, Nasrin Rasgoo, Shadi Amin, Nasrin Rastgoo, Mehdi Arbabi
Rok vydání: 2011
Předmět:
medicine.medical_treatment
polymerase chain reaction
enhanced green fluorescent protein
Mice
Carcinoembryonic antigen
Neoplasms
binding affinity
Immunology and Allergy
Cloning
Molecular
cancer cell
Mice
Inbred BALB C
biology
Tumor Markers
Biological
Monoclonal
Immunotherapy
Antibody
in vitro study
DNA
Complementary
medicine.drug_class
Immunology
animal experiment
Green Fluorescent Proteins
Molecular Sequence Data
Enzyme-Linked Immunosorbent Assay
Monoclonal antibody
medicine
Biomarkers
Tumor
Escherichia coli
Animals
Humans
Amino Acid Sequence
chimeric protein
mouse
Tumor marker
Hybridomas
Base Sequence
human cell
Cancer
Sequence Analysis
DNA
medicine.disease
Virology
Molecular biology
enzyme linked immunosorbent assay
Carcinoembryonic Antigen
Spectrometry
Fluorescence
monoclonal antibody
biology.protein
gene expression
single chain fragment variable antibody
Single-Chain Antibodies
Zdroj: Hybridoma (2005). 30(3)
ISSN: 1557-8348
Popis: According to World Health Organization (WHO), cancer is a leading cause of death worldwide, accounting for 7.4 million deaths (around 13% of all deaths) in 2004. Monoclonal/recombinant antibodies, which specifically target clinical biomarkers of disease, have increasingly been applied as powerful tools in cancer imaging and therapy, a fact that is highlighted by some nine FDA-approved monoclonal antibodies (MAbs) or their immunoconjugates (as of December 2008) for use in cancer treatment. In this study, five monoclonal antibodies (MAbs) were generated and characterized against carcinoembryonic antigen (CEA), which is widely used clinically as both a blood and tissue tumor marker of epithelial malignancy. Variable domains (VH and VL) of one the stable MAbs with highest affinity were PCR-amplified and assembled as single-chain antibody fragment (scFv). Following the cloning and expression of scFv antibody fragments in Escherichia coli, the functional binding and specificity of the recombinant antibody were confirmed by ELISA. To develop a direct in vitro detection of CEA-positive cancer cells, scFv DNA was genetically fused to enhanced green fluorescent protein (EGFP) gene and expressed in bacteria. The chimeric fluorescent protein is able to specifically detect CEA-positive cell lines; no cross-reactivity was observed with a negative control cell line. This strategy will likely allow the establishment of a rapid, single-step detection assay of CEA, which is considered to be one of the best predictors of malignancy among all other tumor markers. © Copyright 2011, Mary Ann Liebert, Inc.
Databáze: OpenAIRE
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