title: Production of recombinant scFv against p24 of human immunodeficiency virus type 1 by phage display technology.
abstract: Phage display has a fundamental role in protein isolation and engineering. Isolated proteins produced with this method can be modified for specific binding and affinity. P24 is the most produced protein during human immune deficiency virus (HIV) replication; especially in the early steps of HIV-1 infection, its evaluation may have diagnostic values. To test the HIV-1 infection, p24 antigen assay appears to be a very promising alternative to RNA assays. In this study, we have generated a recombinant mouse single chain antibody fragment against p24 of the HIV-1 with the use of phage display technology. After isolation of antibody variable-region (V) gene of B cells extracted from the spleen of an immunized mouse, a library of single chain Fv fragments (scFv) was constructed. The library was used in a series of bio-panning processes against recombinant p24 protein expressed from Escherichia coli. The isolated scFv antibody specifically recognizes the HIV-1 capsid protein p24. The affinity constant of the isolated scFv antibody (MF85) was found to be 2×10(-9) M. Our studies showed that the MF85 scFV antibody has similar properties as that of monoclonal antibodies produced by the hybridoma technology.
authors.affiliation: 1 Medical Biotechnology Department, Faculty of Medical Sciences, Tarbiat Modares University , Tehran, Iran .
authors.forename: Sara
authors.initials: S
authors.lastname: Mohammadzadeh
authors.forename: Masoumeh
authors.initials: M
authors.lastname: Rajabibazl
authors.forename: Mehdi
authors.initials: M
authors.lastname: Fourozandeh
authors.forename: Mohammad Javad
authors.initials: MJ
authors.lastname: Rasaee
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Rahbarizadeh
authors.forename: Mohammad
authors.initials: M
authors.lastname: Mohammadi
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D061505:Cell Surface Display Techniques / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D015658:HIV Infections / Q000175:diagnosis / Q000473:pathology; D015497:HIV-1 / Q000276:immunology; D006801:Humans; D051379:Mice; D019151:Peptide Library; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D015682:Retroviridae Proteins, Oncogenic / Q000276:immunology*; D057127:Single-Chain Antibodies / Q000096:biosynthesis / Q000276:immunology*
chemical_list: D017931:DNA Primers; D019151:Peptide Library; D011994:Recombinant Proteins; D015682:Retroviridae Proteins, Oncogenic; D057127:Single-Chain Antibodies; C060954:p24 protein, Human T-lymphotropic virus 1