Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
Poult Sci. 2022 Mar;101(3):101631. doi: 10.1016/j.psj.2021.101631. Epub 2021 Dec 1.
Growth hormone (GH) is required for normal postnatal development in poultry; however, no immunoassay exists to assess its levels in geese plasma, hindering the study of endocrine regulation in this species. We developed a sandwich ELISA to determine the GH concentrations in the plasma of geese. Recombinant goose GH was produced using a eukaryotic expression system and purified for use as the reference standard in ELISA and the antigen for producing the polyclonal antibodies in rabbits. Rabbit anti-goose GH polyclonal antibody was used to coat the wells of the ELISA plate, and its biotinylated form served as the detection antibody. An avidin-conjugated horseradish peroxidase was used to bind the detection antibody and catalyze the chromogenic reaction of 3,3,5,5-tetramethylbenzidine and HO. A sigmoidal curve was fitted to the optical density and the log of the standard GH concentration using the four-parameter logistic model. The sensitivity of the assay was less than 0.156 ng/mL. The intra- and interassay coefficients of variation were less than 9 and 13%, respectively. The response curve of the serially diluted plasma samples from geese exhibited a good parallel relationship with that observed for the reference standards. The assay effectively detected differences in GH concentrations in plasma samples from geese at various physiological stages; thus, it will be useful for future study of their growth and metabolism.
生长激素（GH）是禽类正常产后发育所必需的；然而，目前还没有免疫测定法来评估鹅血浆中的 GH 水平，这阻碍了对该物种内分泌调节的研究。我们开发了一种夹心 ELISA 来测定鹅血浆中的 GH 浓度。使用真核表达系统生产重组鹅 GH，并将其纯化，用作 ELISA 的参考标准和制备兔多克隆抗体的抗原。兔抗鹅 GH 多克隆抗体用于包被 ELISA 板的孔，其生物素化形式用作检测抗体。亲和素结合的辣根过氧化物酶用于结合检测抗体并催化 3,3,5,5-四甲基联苯胺和 HO 的显色反应。使用四参数逻辑模型拟合光密度和标准 GH 浓度对数的 S 形曲线。该测定法的灵敏度小于 0.156ng/mL。批内和批间变异系数分别小于 9%和 13%。来自不同生理阶段鹅的连续稀释血浆样品的反应曲线与参考标准观察到的曲线具有良好的平行关系。该测定法有效地检测了不同生理阶段鹅血浆样品中 GH 浓度的差异；因此，它将有助于未来对其生长和代谢的研究。
