PMID: 8066995May 1, 1994

Preparation of anti-canine serum amyloid A (SAA) serum and purification of SAA from canine high-density lipoprotein

Veterinary Immunology and Immunopathology
S YamamotoY Rikihisa


Antiserum to canine serum amyloid A (SAA) was prepared in rabbits by immunization with crude SAA which was prepared from high-density lipoprotein 3 (HDL3) obtained from canine acute-phase serum. The antiserum was absorbed for contaminating antibodies by affinity chromatography using Sepharose 4B coupled with normal canine serum proteins. The rabbit anti-canine SAA serum reacted with a protein and formed a single precipitin line at the position of the alpha 1-region of the immunoelectrophoresis of canine acute-phase serum but did not react with the normal canine serum on immunoelectrophoresis. The antibody to canine SAA was also confirmed by Western blotting analysis. Canine SAA was purified as a low molecular weight protein component from crude SAA by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) after gel filtration chromatography. Purified canine SAA had a molecular weight of 15,000 as estimated by SDS-PAGE. This SAA level was found by enzyme-linked immunosorbent assay (ELISA) to increase 1 day after inoculation with Bordetella bronchiseptica to 9.0-20.1 times the preinoculation value.


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