title: Generation and Identification of Peptide-Based Monoclonal Antibodies Against Vacuolar Proton Pyrophosphatase of Toxoplasma gondii.
abstract: Vacuolar proton pyrophosphatase (V-PPase), an electrogenic proton pump widely distributed in non-mammalian species, is one of the important targets for acidocalcisomes. In this study, a novel method of peptide-based antibody generation was performed to produce monoclonal antibodies (MAbs) against Toxoplasma gondii V-PPase. Three hybridomas were identified and confirmed by ELISA, Western blotting, and immunofluorescence. All of them can react with an 85 kDa band of T. gondii protein in purified acidocalcisomal fraction. The three MAbs were all specific to the synthetic peptide of YTKAADVGADLSGKNEYGMSEDDPRNPAC, corresponding to amino acids at the location of 292aa-320aa of TgVP1 amino acid sequence. These specific MAbs will be valuable tools for further study of T. gondii infection biology, pathogenesis, and host immune response.
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, Guangdong, P.R. China .
authors.forename: Chengbi
authors.initials: C
authors.lastname: Tong
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, Guangdong, P.R. China .
authors.forename: Bin
authors.initials: B
authors.lastname: Xiao
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, Guangdong, P.R. China .
authors.forename: Shasha
authors.initials: S
authors.lastname: Cheng
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, Guangdong, P.R. China .
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, Guangdong, P.R. China .
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Liao
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, Guangdong, P.R. China .
authors.forename: Shuhong
authors.initials: S
authors.lastname: Luo
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, Guangdong, P.R. China .
authors.forename: Wenbo
authors.initials: W
authors.lastname: Hao
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000917:Antibody Formation; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D010446:Peptide Fragments / Q000276:immunology*; D015800:Protozoan Proteins / Q000276:immunology*; D011755:Pyrophosphatases / Q000276:immunology*; D014122:Toxoplasma / Q000201:enzymology*; D014617:Vacuoles / Q000201:enzymology*
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D010446:Peptide Fragments; D015800:Protozoan Proteins; D011755:Pyrophosphatases