title: Generation and characterization of anti-chitinase monoclonal antibodies.
abstract: Class IV chitinase, an allergenic protein of Vitis vinifera (grape), was purified by anion exchange chromatography and used for immunization of Balb/c mice. Monoclonal antibodies (MAbs) were raised by hybridoma technology using Sp2/0 myeloma cells. Finally after three limiting dilutions, six stable clones were generated. Antibody isotyping showed that IgG(2a), IgG(2b), and IgM were produced by one, two, and three of the clones, respectively. All of the MAbs had kappa light chain. The affinities were in the range of 3 × 10(8) to 1.2 × 10(9) M(-1). The MAbs were specific for grape chitinase as confirmed by Western blotting. In conclusion, we successfully produced several MAbs against grape class IV chitinase, which could be used for assessment of this allergen in different grape cultivars.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Khorasan Razavi, Islamic Republic of Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Soukhtanloo
authors.forename: Reza
authors.initials: R
authors.lastname: Falak
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Sankian
authors.forename: A-Reza
authors.initials: AR
authors.lastname: Varasteh
mesh_terms: D000485:Allergens / Q000276:immunology* / Q000302:isolation & purification; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification*; D000915:Antibody Affinity; D000918:Antibody Specificity; D015153:Blotting, Western; D002688:Chitinases / Q000276:immunology* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005638:Fruit / Q000737:chemistry / Q000201:enzymology / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D007114:Immunization; D007132:Immunoglobulin Isotypes / Q000032:analysis; D007145:Immunoglobulin kappa-Chains / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D009101:Multiple Myeloma; D010936:Plant Extracts / Q000737:chemistry / Q000276:immunology*; D027843:Vitis / Q000737:chemistry / Q000201:enzymology / Q000276:immunology*
chemical_list: D000485:Allergens; D000911:Antibodies, Monoclonal; D007132:Immunoglobulin Isotypes; D007145:Immunoglobulin kappa-Chains; D010936:Plant Extracts; D002688:Chitinases