Abstract
Bovine AA amyloidosis is the most frequently encountered amyloid type in cattle, and it is characterized by an extracellular deposition of pathological amyloid A (AA) protein. Because of the lack of a specific monoclonal antibody (mAbs) against bovine amyloid A (bAA) protein and its precursor, bovine serum amyloid A1 (bSAA1), at present anti-bAA rabbit antiserum and anti-human AA or SAA mAbs are widely used for diagnosis and analysis of bovine AA amyloidosis. In this study, three specific mAbs against bSAA1 were isolated by immunization using synthetic peptides of bSAA1, and these mAbs showed higher detection ability and specificity to bAA and bSAA1 than rabbit antiserum and anti-human AA or SAA mAbs in Western blot analysis and immunohistochemistry. These novel mAbs will be valuable in the development of a more precise immunochemical diagnostic tool for bovine AA amyloidosis, as well as for studying the pathophysiological mechanisms involved in this disease.
Abbreviations | ||
AA | = | amyloid A |
aa | = | amino acid |
AEF | = | amyloid enhancing factor |
bAA | = | bovine AA |
bSAA | = | bovine serum amyloid A |
ELISA | = | enzyme-linked immunosorbent assay |
HRP | = | horseradish peroxidase |
mAbs | = | monoclonal antibodies |
MI | = | medullary interstitium |
PBS | = | phosphate buffered saline |
PBST | = | PBS containing 0.1% Tween 20 |
rbSAA | = | recombinant bSAA |
RT-PCR | = | reverse transcription-polymerase chain reaction |
SAA | = | serum amyloid A |
SDS | = | sodium dodecyl sulfate |
TBM | = | tubular basement membrane |
TEC | = | tubular epithelial cytoplasm |
Abbreviations | ||
AA | = | amyloid A |
aa | = | amino acid |
AEF | = | amyloid enhancing factor |
bAA | = | bovine AA |
bSAA | = | bovine serum amyloid A |
ELISA | = | enzyme-linked immunosorbent assay |
HRP | = | horseradish peroxidase |
mAbs | = | monoclonal antibodies |
MI | = | medullary interstitium |
PBS | = | phosphate buffered saline |
PBST | = | PBS containing 0.1% Tween 20 |
rbSAA | = | recombinant bSAA |
RT-PCR | = | reverse transcription-polymerase chain reaction |
SAA | = | serum amyloid A |
SDS | = | sodium dodecyl sulfate |
TBM | = | tubular basement membrane |
TEC | = | tubular epithelial cytoplasm |
Acknowledgements
We acknowledge Dr. H. Sakai (Gifu University) for his assistance with the immunohistochemistry. This study was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan.