title: The use of Nanotrap particles technology in capturing HIV-1 virions and viral proteins from infected cells.
abstract: HIV-1 infection results in a chronic but incurable illness since long-term HAART can keep the virus to an undetectable level. However, discontinuation of therapy rapidly increases viral burden. Moreover, patients under HAART frequently develop various metabolic disorders and HIV-associated neuronal disease. Today, the main challenge of HIV-1 research is the elimination of the residual virus in infected individuals. The current HIV-1 diagnostics are largely comprised of serological and nucleic acid based technologies. Our goal is to integrate the nanotrap technology into a standard research tool that will allow sensitive detection of HIV-1 infection. This study demonstrates that majority of HIV-1 virions in culture supernatants and Tat/Nef proteins spiked in culture medium can be captured by nanotrap particles. To determine the binding affinities of different baits, we incubated target molecules with nanotrap particles at room temperature. After short sequestration, materials were either eluted or remained attached to nanotrap particles prior to analysis. The unique affinity baits of nanotrap particles preferentially bound HIV-1 materials while excluded albumin. A high level capture of Tat or Tat peptide by NT082 and NT084 particles was measured by western blot (WB). Intracellular Nef protein was captured by NT080, while membrane-associated Nef was captured by NT086 and also detected by WB. Selective capture of HIV-1 particles by NT073 and NT086 was measured by reverse transcriptase assay, while capture of infectious HIV-1 by these nanoparticles was demonstrated by functional transactivation in TZM-bl cells. We also demonstrated specific capture of HIV-1 particles and exosomes-containing TAR-RNA in patients' serum by NT086 and NT082 particles, respectively, using specific qRT-PCR. Collectively, our data indicate that certain types of nanotrap particles selectively capture specific HIV-1 molecules, and we propose to use this technology as a platform to enhance HIV-1 detection by concentrating viral proteins and infectious virions from infected samples.
authors.affiliation: National Center for Biodefense & Infectious Diseases, George Mason University, Manassas, Virginia, United States of America.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Jaworski
authors.affiliation: National Center for Biodefense & Infectious Diseases, George Mason University, Manassas, Virginia, United States of America.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Saifuddin
authors.affiliation: National Center for Biodefense & Infectious Diseases, George Mason University, Manassas, Virginia, United States of America.
authors.forename: Gavin
authors.initials: G
authors.lastname: Sampey
authors.affiliation: National Center for Biodefense & Infectious Diseases, George Mason University, Manassas, Virginia, United States of America.
authors.forename: Nazly
authors.initials: N
authors.lastname: Shafagati
authors.affiliation: National Center for Biodefense & Infectious Diseases, George Mason University, Manassas, Virginia, United States of America; Department of Microbiology, Immunology, and Tropical Medicine, George Washington University, Washington, District of Columbia, United States of America.
authors.forename: Rachel
authors.initials: R
authors.lastname: Van Duyne
authors.affiliation: National Center for Biodefense & Infectious Diseases, George Mason University, Manassas, Virginia, United States of America.
authors.forename: Sergey
authors.initials: S
authors.lastname: Iordanskiy
authors.affiliation: National Center for Biodefense & Infectious Diseases, George Mason University, Manassas, Virginia, United States of America.
authors.forename: Kylene
authors.initials: K
authors.lastname: Kehn-Hall
authors.affiliation: Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, United States of America.
authors.forename: Lance
authors.initials: L
authors.lastname: Liotta
authors.affiliation: Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, United States of America.
authors.forename: Emanuel
authors.initials: E
authors.lastname: Petricoin
authors.affiliation: Division of Infectious Diseases, Georgetown University, Washington, District of Columbia, United States of America.
authors.forename: Mary
authors.initials: M
authors.lastname: Young
authors.affiliation: Ceres Nanosciences, Manassas, Virginia, United States of America.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Lepene
authors.affiliation: National Center for Biodefense & Infectious Diseases, George Mason University, Manassas, Virginia, United States of America.
authors.forename: Fatah
authors.initials: F
authors.lastname: Kashanchi
mesh_terms: D002460:Cell Line; D015696:Gene Products, tat / Q000032:analysis; D015658:HIV Infections / Q000175:diagnosis*; D015497:HIV-1 / Q000737:chemistry* / Q000472:pathogenicity*; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry*; D014764:Viral Proteins / Q000032:analysis*; D014771:Virion / Q000737:chemistry*; D054311:nef Gene Products, Human Immunodeficiency Virus / Q000032:analysis
chemical_list: D015696:Gene Products, tat; D014764:Viral Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus