title: Molecular characterization and validation of commercially available methods for haptoglobin measurement in bottlenose dolphin.
abstract: Haptoglobin (Hp) is a positive acute-phase protein and a valuable marker of inflammation in both human and veterinary medicine. The aim of this study was to validate the molecular characterization of Hp in dolphins and to validate commercially available Hp measurement methods such as Hp-ELISA (originally designed for pigs) and Hp-hemoglobin (Hb) binding assay. The dolphin Hp (dHp) amino acid sequence appeared most similar to pig Hp by sequence homology and phylogenetic clustering. Amino acid sequence analysis revealed that dHp comprises the Hp1 form of α1 and β chains. The anti-pig Hp antibody cross-reacted with both recombinant dHp, expressed by Escherichia coli, and dHp from serum. The intra- and inter-assay levels of imprecision of pig Hp-ELISA and the Hp-Hb binding assay were found to be tolerable for the determination of Hp in dolphin, and there was no significant discrepancy between the two determination methods. The ability of the assay to differentiate between healthy and inflammation groups was investigated, and a significant increase in Hp concentration was detected in inflammatory conditions. Thus, Hp is a useful inflammation marker for dolphin, and the Hp concentration in dolphin serum samples can be reliably measured using commercially available pig Hp-ELISA and Hp-Hb binding assay.
authors.affiliation: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Segawa
authors.affiliation: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Hazumu
authors.initials: H
authors.lastname: Amatsuji
authors.affiliation: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Kento
authors.initials: K
authors.lastname: Suzuki
authors.affiliation: Department of Marine Science and Resources, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Miwa
authors.initials: M
authors.lastname: Suzuki
authors.affiliation: Okinawa Churaumi Aquarium, Motobu-Cho, Okinawa 905-0206, Japan.
authors.forename: Makio
authors.initials: M
authors.lastname: Yanagisawa
authors.affiliation: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Itou
authors.affiliation: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Takeo
authors.initials: T
authors.lastname: Sakai
authors.affiliation: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Nakanishi
keywords: Dolphin; ELISA; Haptoglobin; Haptoglobin–hemoglobin binding assay; Validation