title: Ultrasensitive immuno-detection using viral nanoparticles with modular assembly using genetically-directed biotinylation.
abstract: We report a novel, modular approach to immuno-detection based on antibody recognition and PCR read-out that employs antibody-conjugated bacteriophage and easily-manipulated non-pathogenic viruses as affinity agents. Our platform employs phage genetically tagged for in vivo biotinylation during phage maturation that can easily be linked, through avidin, to any biotinylated affinity agent, including full-length antibodies, peptides, lectins or aptamers. The presence of analyte is reported with high sensitivity through real-time PCR. This approach avoids the need to clone antibody-encoding DNA fragments, allows the use of full-length, high affinity antibodies and, by having DNA reporters naturally encapsulated inside the bacteriophage, greatly reduces nonspecific binding of DNA. We validate the efficacy of this new approach through the detection of Vascular Endothelial Growth Factor, a known angiogenic cancer biomarker protein, at attomolar concentrations in bronchoalveolar lavage fluid.
authors.affiliation: Department of Biomedical Engineering, University of Houston, 4800 Calhoun Rd., Houston, TX, 77204, USA, julialitvinov@gmail.com.
authors.forename: Julia
authors.initials: J
authors.lastname: Litvinov
authors.forename: Anna E V
authors.initials: AE
authors.lastname: Hagström
authors.forename: Yubitza
authors.initials: Y
authors.lastname: Lopez
authors.forename: Meenu
authors.initials: M
authors.lastname: Adhikari
authors.forename: Katerina
authors.initials: K
authors.lastname: Kourentzi
authors.forename: Ulrich
authors.initials: U
authors.lastname: Strych
authors.forename: Federico A
authors.initials: FA
authors.lastname: Monzon
authors.forename: William
authors.initials: W
authors.lastname: Foster
authors.forename: Philip T
authors.initials: PT
authors.lastname: Cagle
authors.forename: Richard C
authors.initials: RC
authors.lastname: Willson
mesh_terms: D000914:Antibodies, Viral / Q000378:metabolism; D001435:Bacteriophages / Q000235:genetics*; D019921:Biotinylation; D006801:Humans; D007118:Immunoassay / Q000379:methods; D037102:Lectins / Q000378:metabolism; D053758:Nanoparticles; D010455:Peptides / Q000378:metabolism; D011485:Protein Binding; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis
chemical_list: D000914:Antibodies, Viral; D037102:Lectins; D010455:Peptides; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A