FSH PROCESS CORE

FSH工艺核心实验室(核心B)的目标是提供仪器、支持和培训 在蛋白质化学技术中适应于糖蛋白的表征。这个核心实验室拥有广泛的 促性腺激素的纯化和鉴定经验。促性腺激素起着中枢作用。 在调节生殖系统方面的作用。它们被释放是对间歇性刺激的反应 下丘脑神经元活动协调内源性启动的卵泡发育后期 性腺周期。反过来，性腺激素，特别是类固醇激素雌二醇，反馈给 促性腺激素的合成和翻译后加工。当使用类固醇激素时 在避孕治疗中，促性腺激素是用来治疗不孕症的。其核心是 将为拟议的项目提供三个支持功能，如下所示：功能#1.产生hFSH 糖花。Bin博士将表达二糖化、四糖化和其他hFSH糖型 在哺乳动物细胞系中。帅博士将向博士提供含有FSH的条件培养液。 布特涅夫。功能#2.纯化hFSH糖形制剂并对其进行化学表征。弗拉基米尔博士 布特涅夫将提纯hFSH糖体并对其进行化学表征。这将包括分离hFSH糖形式。 从脑垂体提取液和条件培养液中，测定化学纯度，测定蛋白质 数量，评估蛋白质的完整性，以及确定碳水化合物组成。卵泡刺激素特性 对于解释项目1-3中进行的生物学研究至关重要。卵泡刺激素糖型质控数据 将每月发布在Core C维护的Glycomics数据共享页面上。功能#3。提供 为项目研究人员提供卵泡刺激素测定和细胞培养支持。杰弗里·V·梅博士将表演 猪颗粒细胞原代培养中的受体结合和类固醇激素合成试验 纯化的hFSH‘糖体的生物活性确认系统。然后，这些准备工作将被释放到 项目调查员。这些化验也将作为一项服务提供给项目调查人员。项目 1和2将在一个普通的细胞模型中进行一些实验。最初，这将是猪的颗粒细胞。 梅博士将评估可用的人类颗粒细胞系。例如，项目调查员 浪费了宝贵的时间来确定KGN细胞，一个假想的人类颗粒细胞系，不适合 我们的学习。如果这个项目得到资助，那么核心B将评估细胞系，并将制造合适的细胞系 可供项目使用。这一核心还将为单抗的表达提供支持 项目1.梅博士将根据他在原代细胞培养方面的经验担任全面的核心领导 如蛋白质化学和分子生物学。

The goal of the FSH process core laboratory (Core B) is to provide instumentation, support and training in protein chemistry techniques adapted to glycoprotein characterization. This core laboratory has extensive experience with gonadotropin purification and characterization. The gonadotropic hormones play a central role in regulating the reproductive system. They are released in response to episodic stimulation by hypothalamic neuronal activity to coordinate the later stages of follicle development initiated by endogenous gonadal cycles. In turn, gonadal hormones, particularly the steroid hormone estradiol feed back to the pituitary and alter gonadotropin synthesis and posttranslational processing. While steroid hormones are used therapeutically in contraceptive procedures, it is the gonadotropins that are used to treat infertility. The core will provide three support functions for the proposed project, as follows: Function #1. Produce hFSH glycofoms. Di-glycosylated, tetra-glycosylated and other hFSH glycoforms will be expressed by Dr. Bin Shuai in mammalian cell lines. Dr. Shuai will provide FSH-containng conditioned culture medium to Dr. Butnev. Function #2. Purify and chemically characterize hFSH glycoform preparations. Dr. Vladimir Butnev will purify and chemically characterize hFSH glycoforms. This will include isolating hFSH glycoforms from pituitary extracts and conditioned culture meduim, determining chemical purity, determining protein quantity, assessing protein integrity, as well as determining carbohydrate composition. FSH characterization is critical for interpreting the biological studies performed in Projects 1-3. FSH glycoform quality control data will be posted monthly on the Glycomics data-sharing page maintained by Core C. Function #3. Provide FSH assay and cell culture support for Project researchers. Dr. Jeffrey V. May will perform receptor-binding and steroidogenic assays in the well-characterized porcine granulosa cell primary culture system to confirm biological activity of purified hFSH'glycoforms. The preparations will then be released to project investigators. These assays will also be made available to project investigators as a service. Projects 1 and 2 will perform some experiments in a common cell model. Initially, this will be porcine granulosa cells. Dr. May will evaluate human granulosa cell lines that become available. For example project investigators wasted valuable time determining that KGN cells, a supposed human granulosa cell line, were unsuitable for our studies. If this project is funded, then Core B will evaluate cell lines and suitable lines will be made available to the projects. This core will also provide support for monoclonal antibody expression to support project 1. Dr. May will serve as overall Core Leader based upon his experience in primary cell culture as well as protein chemistry and molecular biology.