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使用抗人促卵泡激素β亚基单克隆抗体开发一种用于测量绢毛猴灵长类动物促卵泡激素(FSH)的酶免疫测定法(EIA)。

Development of an enzyme-immunoassay (EIA) for the measurement of follicle-stimulating-hormone (FSH) in Callitrichid primates using a monoclonal antibody against the human-FSH-beta-subunit.

作者信息

Rosenbusch J, Dias J A, Hodges J K

机构信息

Department of Reproductive Biology, German Primate Center, Goettingen, Germany.

出版信息

Am J Primatol. 1997;41(3):179-93. doi: 10.1002/(SICI)1098-2345(1997)41:3<179::AID-AJP2>3.0.CO;2-U.

Abstract

Despite the importance of Callitrichid primates in both biomedical and conservation research, practical and reliable immunoassays for the measurement of follicle-stimulating hormone (FSH) have not yet been described. A panel of monoclonal antibodies against specific peptide fragments within either the alpha or beta subunit of human FSH was evaluated for their ability to recognize FSH from Callitrichid and other New World primates. One of these, monoclonal antibody 46.3h6.b7 raised against human FSH, was selected due to its ability to recognize marmoset monkey FSH and its low crossreactivity with other gonadotrophins. The antibody formed the basis of an enzymeimmunoassay using a highly purified human urinary FSH (Metrodin, Serono) preparation coupled to biotin as label and unmodified as standard. After 24 h incubation, antibody bound label was visualized by addition of streptavidin-peroxidase followed by the appropriate substrate. Parallelism was obtained between the standard and dilutions of pituitary extracts, urine and plasma from the common marmoset (Callithrix jacchus) as well as from two tamarin species (Saguinus fuscicollis and S. oedipus) and one squirrel monkey (Saimiri sciureus). Profiles of plasma and urinary FSH during the follicular phase are shown for two individual marmosets. The ability to measure FSH in Callitrichidae provides new opportunities for studies of the reproductive biology of these New World primate species.

摘要

尽管绢毛猴灵长类动物在生物医学和保护研究中都很重要,但尚未有用于测量促卵泡激素(FSH)的实用且可靠的免疫测定方法被描述。评估了一组针对人FSHα或β亚基内特定肽段的单克隆抗体识别绢毛猴科和其他新大陆灵长类动物FSH的能力。其中一种,即针对人FSH产生的单克隆抗体46.3h6.b7,因其能够识别狨猴FSH且与其他促性腺激素的交叉反应性低而被选中。该抗体构成了一种酶免疫测定法的基础,该方法使用与生物素偶联作为标记且未修饰作为标准品的高度纯化的人尿FSH(Metrodin,雪兰诺公司)制剂。孵育24小时后,通过添加链霉亲和素 - 过氧化物酶,然后添加适当的底物来使结合了标记的抗体可视化。普通狨猴(Callithrix jacchus)以及两种柽柳猴(Saguinus fuscicollis和S. oedipus)和一只松鼠猴(Saimiri sciureus)的垂体提取物、尿液和血浆的标准品与稀释液之间呈现出平行关系。展示了两只个体狨猴卵泡期血浆和尿FSH的变化曲线。测量绢毛猴科动物FSH的能力为研究这些新大陆灵长类物种的生殖生物学提供了新的机会。

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