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Development and characterization of monoclonal antibodies against canine trypsin |
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| Authors: | Takaki Waritani Yoko Okuno Yoshinori Ashida Ryo Tsuchiya Kosaku Kobayashi Takatsugu Yamada |
| Institution: | a Animal Health Products and Chemicals Division, Animal Health Products and Chemicals Research Section, Fuji Chemical Industries Ltd., 530 Chokeiji, Takaoka-shi, Toyama 933-8511, Japan b Laboratory of Veterinary Internal Medicine, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Sagamihara-shi, Kanagawa 229-8501, Japan |
| Abstract: | Canine cationic trypsin was purified by salting-out, gel filtration and affinity chromatography. Purity was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight was ca. 28 kDa by SDS-PAGE. Thirty hybridomas were obtained which produced mAb to canine cationic trypsin by the cell fusion technique. Twenty-two of these recognized cationic trypsin only, while eight hybridomas recognized both cationic and anionic trypsin. Several of the anti-canine cationic trypsin mAb were purified by salting-out and DEAE ion-change chromatography using ascites fluid of immunized BALB/c mice. The mAb proved to have very high specificity to canine cationic trypsin as shown by immunoblotting and it may be possible to use them to develop clinical assays. |
| Keywords: | Dog Monoclonal antibodies Trypsin Trypsin-like immunoreactivity |
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