AIMS: This study aims to develop and characterize monoclonal antibodies (Mabs) with high specificity and affinity for surface antigens of an epidemiologically important serotype 4b of Listeria monocytogenes. METHODS AND RESULTS: Hybridoma clones were derived from B lymphocytes of mice immunized with L. monocytogenes serotype 4b and screened against this strain by an enzyme-linked immunosorbent assay. Twenty-nine clones secreting Mabs reactive with formalin-killed bacteria were obtained; 15, 8, 5 and 1 Mabs were immunoglobulin subclasses IgG2a, IgG2b, IgM and IgG1, respectively. Immunofluorescence or immunogold labelling demonstrated all except five IgM and one IgG2a Mabs bound to the surface of a live L. monocytogenes serotype 4b. The majority of the 23 surface-binding Mabs recognized linear epitopes on a 77-kDa protein. These surface-binding Mabs exhibited little or no cross-reactivity with non-4b serotypes (1/2a, 1/2b, 3a, etc.) of L. monocytogenes, five other Listeria species, Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium. CONCLUSIONS: The Mabs recognizing a 77-kDa surface protein are novel antibodies with specificity and affinity for L. monocytogenes serotype 4b. SIGNIFICANCE AND IMPACT OF THE STUDY: These anti-77 kDa surface protein Mabs may be explored as reagents for the development of Mabs-based diagnostic immunoassays for L. monocytogenes serotype 4b strains.
AIMS: This study aims to develop and characterize monoclonal antibodies (Mabs) with high specificity and affinity for surface antigens of an epidemiologically important serotype 4b of Listeria monocytogenes. METHODS AND RESULTS: Hybridoma clones were derived from B lymphocytes of mice immunized with L. monocytogenes serotype 4b and screened against this strain by an enzyme-linked immunosorbent assay. Twenty-nine clones secreting Mabs reactive with formalin-killed bacteria were obtained; 15, 8, 5 and 1 Mabs were immunoglobulin subclasses IgG2a, IgG2b, IgM and IgG1, respectively. Immunofluorescence or immunogold labelling demonstrated all except five IgM and one IgG2a Mabs bound to the surface of a live L. monocytogenes serotype 4b. The majority of the 23 surface-binding Mabs recognized linear epitopes on a 77-kDa protein. These surface-binding Mabs exhibited little or no cross-reactivity with non-4b serotypes (1/2a, 1/2b, 3a, etc.) of L. monocytogenes, five other Listeria species, Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium. CONCLUSIONS: The Mabs recognizing a 77-kDa surface protein are novel antibodies with specificity and affinity for L. monocytogenes serotype 4b. SIGNIFICANCE AND IMPACT OF THE STUDY: These anti-77 kDa surface protein Mabs may be explored as reagents for the development of Mabs-based diagnostic immunoassays for L. monocytogenes serotype 4b strains.
Authors: Teela Boivin; Cathie Elmgren; Brian W Brooks; Hongsheng Huang; Franco Pagotto; Min Lin Journal: Appl Environ Microbiol Date: 2016-10-27 Impact factor: 4.792
Authors: Eric T Sumrall; Christian Röhrig; Mario Hupfeld; Lavanja Selvakumar; Jiemin Du; Matthew Dunne; Mathias Schmelcher; Yang Shen; Martin J Loessner Journal: Appl Environ Microbiol Date: 2020-06-17 Impact factor: 4.792
Authors: Marcelo Mendonça; Neida L Conrad; Fabricio R Conceição; Angela N Moreira; Wladimir P da Silva; José Ag Aleixo; Arun K Bhunia Journal: BMC Microbiol Date: 2012-11-23 Impact factor: 3.605