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B-cell epitope specificity of carboxy terminus of Mycobacterium paratuberculosis ModD

Authors
 Donghee Cho  ;  Sung Jae Shin  ;  Michael T. Collins 
Citation
 JOURNAL OF IMMUNOASSAY & IMMUNOCHEMISTRY , Vol.31(3) : 181-192, 2010 
Journal Title
 JOURNAL OF IMMUNOASSAY & IMMUNOCHEMISTRY 
ISSN
 1532-1819 
Issue Date
2010
MeSH
Animals ; Antibody Specificity* ; Antigen-Antibody Reactions ; Antigens, Bacterial/chemistry* ; Antigens, Bacterial/immunology* ; Antigens, Bacterial/isolation & purification ; Cattle ; Enzyme-Linked Immunosorbent Assay ; Epitope Mapping ; Epitopes, B-Lymphocyte/immunology* ; Epitopes, B-Lymphocyte/isolation & purification ; Mycobacterium avium subsp. paratuberculosis/chemistry* ; Mycobacterium avium subsp. paratuberculosis/immunology ; Mycobacterium avium subsp. paratuberculosis/isolation & purification ; Paratuberculosis/immunology ; Peptides/chemical synthesis ; Peptides/chemistry ; Peptides/immunology ; Rabbits ; Sensitivity and Specificity
Keywords
culture filtrate ; epitope ; ModD ; Mycobacterium avium ; Mycobacterium paratuberculosis ; synthesized peptide
Abstract
Epitope mapping of ModD of Mycobacterium paratuberculosis was performed using overlapping peptides. In total, 80 overlapping peptides, covering the entire mature ModD, were commercially synthesized. Each peptide spanned 14 amino acids with an offset of 4 amino acids, i.e., with an overlap of 10 amino acids. Synthetic peptide antigenicity was evaluated by enzyme-linked immunosorbent assay (ELISA) using rabbit antisera to culture filtrate (CF) of M. avium or M. paratuberculosis or recombinant ModD (rModD). The peptides o f ModD reacting most strongly (ELISA OD > 1.0) were clustered near the N- and C-terminal ends. The peptides around the C-terminal end only showed the greatest specificity for M. paratuberculosis, yielding high ELISA OD values with rabbit anti-M. paratuberculosis CF serum and low ELISA OD values with rabbit anti-M. avium CF serum. Sera from naturally M. paratuberculosis-infected cattle, however, bound poorly to the short, 14-amino-acid peptides. Thus, two longer peptides covering amino acids 100 to 125 and 328 to 353 were synthesized based on their broad reactivity to rabbit serum against M. paratuberculosis CF. The peptide covering amino acids 328 to 353 showed the highest level of specific bovine antibody binding
Full Text
http://www.tandfonline.com/doi/full/10.1080/10739149.2010.488607
DOI
10.1080/10739149.2010.488607
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers
Yonsei Authors
Shin, Sung Jae(신성재) ORCID logo https://orcid.org/0000-0003-0854-4582
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/103247
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