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The feasibility and usability of DNA-dot bioconjugation to antibody for targeted in vitro cancer cell fluorescence imaging.
J Photochem Photobiol B. 2020 Jun 26; 209:111944.JP

Abstract

DNA-protein bioconjugation is an appealing target-tracking strategy. The new capability of DNA molecule as a biological nanomaterial endows unique fluorescence and physicochemical properties to be applied in bioimaging. Progression in targeted imaging is contingent on the conjugation of diagnostic nanoparticles to biomolecular signatures, particularly antibody-based ligands. Here, we have reported our recent experience, DNA-dot synthesis and characterization, the covalent conjugation of DNA-dot to goat F(ab')2 IgG and Epidermal Growth Factor Receptor (EGFR) antibodies, DNA-dot@antibody coupling confirmation, and fluorescent targeted imaging of lung cancer cell line. As a result, the average size of DNA-dot was 4.5-5 nm which was conjugated to amine-rich antibodies with returned PO4-1 groups on the DNA-dot surface via PN bond. The synthetic DNA-dots were conjugated to the goat F(ab')2 IgG and tested for fluorescent detection usability by indirect Dot-blot assay. Also, DNA-dot@EGFR conjugates identified lung cancer cells with 84-92% specificity and 100% sensitivity in five concentrations, associated with 0.0025 to 0.04 g 100 μL-1 DNA-dot. The results demonstrated that bioconjugated DNA-dot can do the diagnosis profiling of molecular biomarkers. Generally, DNA-dot bioconjugation with antibody is implemented within two days and biomarker detection takes one day. Consequently, DNA-dot@antibody is potentially a toxic-free, swift, and efficient method of antibody labeling that opens up new horizons in fluorescent nanoimaging in the field of cancer cell detection.

Authors+Show Affiliations

Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.Department of Chemical Engineering, Northeastern University, Boston, MA 02115, USA.Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran; Department of Clinical Biochemistry and Laboratory Medicine, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran. Electronic address: Zarghami@tbzmed.ac.ir.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32619869

Citation

Mohajeri, Nasrin, et al. "The Feasibility and Usability of DNA-dot Bioconjugation to Antibody for Targeted in Vitro Cancer Cell Fluorescence Imaging." Journal of Photochemistry and Photobiology. B, Biology, vol. 209, 2020, p. 111944.
Mohajeri N, Mostafavi E, Zarghami N. The feasibility and usability of DNA-dot bioconjugation to antibody for targeted in vitro cancer cell fluorescence imaging. J Photochem Photobiol B, Biol. 2020;209:111944.
Mohajeri, N., Mostafavi, E., & Zarghami, N. (2020). The feasibility and usability of DNA-dot bioconjugation to antibody for targeted in vitro cancer cell fluorescence imaging. Journal of Photochemistry and Photobiology. B, Biology, 209, 111944. https://doi.org/10.1016/j.jphotobiol.2020.111944
Mohajeri N, Mostafavi E, Zarghami N. The Feasibility and Usability of DNA-dot Bioconjugation to Antibody for Targeted in Vitro Cancer Cell Fluorescence Imaging. J Photochem Photobiol B, Biol. 2020 Jun 26;209:111944. PubMed PMID: 32619869.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The feasibility and usability of DNA-dot bioconjugation to antibody for targeted in vitro cancer cell fluorescence imaging. AU - Mohajeri,Nasrin, AU - Mostafavi,Ebrahim, AU - Zarghami,Nosratollah, Y1 - 2020/06/26/ PY - 2020/02/18/received PY - 2020/06/15/revised PY - 2020/06/22/accepted PY - 2020/7/4/pubmed PY - 2020/7/4/medline PY - 2020/7/4/entrez KW - Antibody KW - Bioconjugation KW - Cancer cell KW - DNA-dot KW - EGFR KW - Fluorescence imaging SP - 111944 EP - 111944 JF - Journal of photochemistry and photobiology. B, Biology JO - J. Photochem. Photobiol. B, Biol. VL - 209 N2 - DNA-protein bioconjugation is an appealing target-tracking strategy. The new capability of DNA molecule as a biological nanomaterial endows unique fluorescence and physicochemical properties to be applied in bioimaging. Progression in targeted imaging is contingent on the conjugation of diagnostic nanoparticles to biomolecular signatures, particularly antibody-based ligands. Here, we have reported our recent experience, DNA-dot synthesis and characterization, the covalent conjugation of DNA-dot to goat F(ab')2 IgG and Epidermal Growth Factor Receptor (EGFR) antibodies, DNA-dot@antibody coupling confirmation, and fluorescent targeted imaging of lung cancer cell line. As a result, the average size of DNA-dot was 4.5-5 nm which was conjugated to amine-rich antibodies with returned PO4-1 groups on the DNA-dot surface via PN bond. The synthetic DNA-dots were conjugated to the goat F(ab')2 IgG and tested for fluorescent detection usability by indirect Dot-blot assay. Also, DNA-dot@EGFR conjugates identified lung cancer cells with 84-92% specificity and 100% sensitivity in five concentrations, associated with 0.0025 to 0.04 g 100 μL-1 DNA-dot. The results demonstrated that bioconjugated DNA-dot can do the diagnosis profiling of molecular biomarkers. Generally, DNA-dot bioconjugation with antibody is implemented within two days and biomarker detection takes one day. Consequently, DNA-dot@antibody is potentially a toxic-free, swift, and efficient method of antibody labeling that opens up new horizons in fluorescent nanoimaging in the field of cancer cell detection. SN - 1873-2682 UR - https://www.unboundmedicine.com/medline/citation/32619869/The_feasibility_and_usability_of_DNA-dot_bioconjugation_to_antibody_for_targeted_in_vitro_cancer_cell_fluorescence_imaging L2 - https://linkinghub.elsevier.com/retrieve/pii/S1011-1344(20)30394-8 DB - PRIME DP - Unbound Medicine ER -
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