title: Peptide-based Polyclonal Antibody Production against P110 Protein of Mycoplasma genitalium.
abstract: Mycoplasma genitalium (M.genitalium) is a sexually transmitted pathogen. Detection of this microorganism in clinical specimens by culture is rather difficult and time consuming. The aim of this study was to produce polyclonal antibody against a synthetic peptide from P110 protein of M.genitalium in order to develop a diagnostic tool for detection of this microorganism in clinical specimens. A synthetic peptide from P110 protein was conjugated to Keyhole Limpet Hemocyanin (KLH) and used for immunization of a white New Zealand rabbit. The produced antibody was purified by affinity chromatography and its specific interaction with immunizing peptide was determined by ELISA. Immunoreactivity of the antibody was also tested by Western blotting in bacterial cell lysate prepared from M.genitalium G-37. To confirm its application as a diagnostic tool, indirect immunofluorescent staining method was performed on M.genitalium-infected PBMC using anti-P110 as the primary antibody. The results showed that produced antibody has excellent reactivity with immunizing peptide and also detected a single band of 110 kDa corresponding to P110 protein. M.genitalium-infected PBMC showed a bright fluorescent signal in IF staining. This antibody might be used as a tool in diagnostic applications.
authors: Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran ; Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
authors: Omid
authors: O
authors: Zarei
authors: Gholam Reza
authors: GR
authors: Irajian
authors: Amir Hassan
authors: AH
authors: Zarnani
authors: Leili
authors: L
authors: Chamani-Tabriz
authors: Shaghayegh
authors: S
authors: Emami
authors: Mahmood
authors: M
authors: Jeddi-Tehrani
authors: Hodjattallah
authors: H
authors: Rabbani
keywords: Antibodies; Mycoplasma genitalium; Peptides