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Amyloid
The Journal of Protein Folding Disorders
Volume 16, 2009 - Issue 4
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Original Article

Isolation and characterization of monoclonal antibodies against bovine serum amyloid A1 protein

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Pages 215-220 | Published online: 19 Nov 2009
 

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.

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