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Research Note: Development of a sandwich ELISA for determining plasma growth hormone concentrations in goose

文献类型： 外文期刊

作者： Chen, R. 1 ; Guo, R. H. 1 ; Lei, M. M. 1 ; Zhu, H. X. 1 ; Yan, L. Y. 1 ; Shi, Z. D. 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Jiangsu Key Lab Food Qual, Minist Sci & Technol, Inst Anim Sci, Nanjing 210014, Peoples R China; Jiangsu Acad Agr Sci, Safety State Key Lab Cultivat Base, Minist Sci & Technol, Inst Anim Sci, Nanjing 210014, Peoples R China

关键词： growth hormone; concentration; sandwich; ELISA; goose

期刊名称：POULTRY SCIENCE （ 影响因子：4.014； 五年影响因子：4.192 ）

ISSN：

年卷期： 2022 年 101 卷 3 期

页码：

收录情况： SCI

摘要： 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 H2O2. 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.

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