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Screening and characterization of monoclonal antibodies to the surface antigens of Listeria monocytogenes serotype 4b
Author(s) -
Lin M.,
Armstrong S.,
Ronholm J.,
Dan H.,
Auclair M.E.,
Zhang Z.,
Cao X.
Publication year - 2009
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2008.04140.x
Subject(s) - listeria monocytogenes , serotype , monoclonal antibody , epitope , microbiology and biotechnology , biology , antibody , antigen , escherichia coli , virology , bacteria , biochemistry , immunology , genetics , gene
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. Immuofluoresence 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.