title: A lateral flow biosensor for rapid detection of DNA-binding protein c-jun.
abstract: A lateral flow biosensor based on an immuno-chromatographic assay has been developed for the detection of DNA-binding proteins. The biosensor is composed of four parts: a sample pad, a conjugate pad, a strip of nitrocellulose membrane and an absorbent pad. A DNA probe containing a specific protein binding consensus sequence is coated onto gold nanoparticles, while an antibody against the DNA-binding protein is immobilized onto a test zone of the nitrocellulose membrane. The target protein binds to the protein binding DNA sequence that is coated on the gold nanoparticles to form nanoparticle-DNA-protein complexes, and the complexes are then captured by the antibody immobilized on the test zone to form a red line for visual detection of the target protein. This biosensor was successfully applied to a DNA-binding protein, c-jun, and the developed biosensor allows for the rapid detection of down to 0.2 footprint unit of c-jun protein within 10 min. This biosensor was verified using HeLa cells and it visually detected c-jun activity in 100 μg of crude cell lysate protein. The antibody against c-jun used in the biosensor can distinguish c-jun from other nonspecific proteins, with high specificity.
authors.affiliation: South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Fang
authors.forename: Chenchen
authors.initials: C
authors.lastname: Ge
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Zhang
authors.forename: Puchang
authors.initials: P
authors.lastname: Lie
authors.forename: Lingwen
authors.initials: L
authors.lastname: Zeng
mesh_terms: D015374:Biosensing Techniques; D002846:Chromatography, Affinity; D015342:DNA Probes / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry; D016755:Proto-Oncogene Proteins c-jun / Q000032:analysis*
chemical_list: D015342:DNA Probes; D016755:Proto-Oncogene Proteins c-jun; D006046:Gold