Document Detail


Production and characterization of monoclonal antibodies detecting chicken interleukin-2 and the development of an antigen capture enzyme-linked immunosorbent assay.
MedLine Citation:
PMID:  11457478     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Eleven monoclonal antibodies (mAbs) which are specific for chicken interleukin-2 (chIL-2) were produced and characterized by enzyme-linked immunosorbent assay (ELISA), Western blotting and neutralizing assays. These mAbs were used to develop a mAb-based antigen capture ELISA specific for chicken IL-2 detection. Anti-IL-2 mAbs bound specifically to E. coli-derived rchIL-2 in ELISA and identified a 16kDa IL-2 polypeptide band in Western blot. Several mAbs were shown to neutralize the biological activities of both rchIL-2 and native chicken IL-2 as measured by concanavalin A (ConA)-induced lymphocyte proliferation assay, IL-2 bioassay, and natural killer cell assay. Among the neutralizing mAbs, the mAb chIL-2/11 was most potent in neutralizing IL-2 activity. To develop a sensitive ELISA for the detection of chicken IL-2, an antigen capture ELISA was developed using the mAb chIL-2/16 as the antigen capture antibody and rabbit anti-IL-2 peptide antibody as the detection antibody. Using the mAb-based antigen capture ELISA, significant correlation between the level of IL-2 detected in bioassays and in ELISA was observed. These results showed that the mAb-based antigen capture ELISA is less time-consuming and more reliable compared to a conventional IL-2 bioassay for chicken IL-2. These neutralizing mAbs will facilitate basic immunobiological studies of the role of IL-2 in normal and disease states in chickens.
Authors:
T Miyamoto; H S Lillehoj; E J Sohn; W Min
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Veterinary immunology and immunopathology     Volume:  80     ISSN:  0165-2427     ISO Abbreviation:  Vet. Immunol. Immunopathol.     Publication Date:  2001 Aug 
Date Detail:
Created Date:  2001-07-17     Completed Date:  2001-09-20     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8002006     Medline TA:  Vet Immunol Immunopathol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  245-57     Citation Subset:  IM    
Affiliation:
BARC-East, Building 1040, Parasite Biology, Epidemiology and Systematics Laboratory, Animal and Natural Resources Institute, USDA/Agricultural Research Service, Beltsville, MD 20705, USA.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Antibodies, Monoclonal / biosynthesis*
Antibody Specificity
Biological Assay
Chickens / immunology*
Enzyme-Linked Immunosorbent Assay / methods*
Interleukin-2 / analysis*,  chemistry,  immunology*
Killer Cells, Natural / immunology
Lymphocyte Activation
Mice
Molecular Weight
Neutralization Tests
Peptide Fragments / chemistry,  immunology
Rabbits
Recombinant Proteins / chemistry,  immunology
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Interleukin-2; 0/Peptide Fragments; 0/Recombinant Proteins

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