Development and characterization of monoclonal antibodies against canine trypsin.
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. 28kDa 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.
Waritani
T
T
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.
Okuno
Y
Y
Ashida
Y
Y
Tsuchiya
R
R
Kobayashi
K
K
Yamada
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D013045:Species Specificity; D014357:Trypsin / Q000276:immunology* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D014357:Trypsin