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号外---景杰生物正式开通外泌体蛋白质组/修饰组学服务啦!!

2017-05-26 景杰生物 景杰生物

编者按:作为蛋白质组学科研服务领域的领跑者,景杰生物一直致力于为客户提供最专业、 深入的蛋白组学/修饰组学的技术服务。我们秉承着“一流人才、一流技术、一流效率”的理念,以“精准医疗”驱动人类健康事业为己任,致力将蛋白质组学技术引入更多的生物学研究领域,服务于有需求的科研工作者。如今我们要开展外泌体的蛋白质组学及修饰组学技术服务啦! 什么是外泌体 外泌体(Exosome)是一种由细胞分泌的

编者按:作为蛋白质组科研服务领域的领跑者,景杰生物一直致力于为客户提供最专业、 深入的蛋白组学/修饰组学的技术服务。我们秉承着“一流人才、一流技术、一流效率”的理念,以“精准医疗”驱动人类健康事业为己任,致力将蛋白质组学技术引入更多的生物学研究领域,服务于有需求的科研工作者。如今我们要开展外泌体的蛋白质组学及修饰组学技术服务啦!

什么是外泌体

外泌体(Exosome)是一种由细胞分泌的膜性囊泡小体,其直径通常为30-100 nm,比其它的一些囊泡(凋亡小体)要小得多。外泌体广泛存在于各种体液中,可携带蛋白质、脂类、核酸(miRNAs,ncRNAs)等多种生物功能大分子。最新的研究表明,外泌体形成了一种特殊的细胞间信息传递系统,它不仅影响细胞的生理状态,而且还与多种疾病的发生与发展密切相关。外泌体领域的研究已经在癌症的诊断及治疗等领域发挥重要作用,基于质谱的高通量蛋白质组学是研究外泌体不可或缺的一项技术手段。

外泌体具有磷脂双分子膜结构,导致其沉降系数和蛋白质聚集体有着很大的不同。而且外泌体膜表面存在特异性膜蛋白如CD9和CD81,前者被证实和肿瘤迁移有关,后者与丙型肝炎病毒的侵染有关。而因为外泌体具有这些显著的易分离的特点,我们可以通过密度梯度离心,亲和层析等方法对外泌体进行分离和纯化。

研究方法

如下图所示,质谱技术可以用于研究外泌体中的成分:蛋白质,脂类和小分子化合物。而利用蛋白质质谱技术,我们可以鉴定外泌体中的蛋白组成并对蛋白进行定量分析。

我们的技术优势

我们拥有Label free、SILAC、TMT/iTRAQ三种技术研究外泌体定量蛋白质组。

Label free的方法无需同位素标记试剂,并且不受样本条件的限制,但准确性会受到质谱重复性等因素的影响;

SILAC技术属于体内标记技术,标记效果稳定,标记效率高达99%,并不受裂解液成分的影响。

TMT/iTRAQ属于体外标记技术,标记灵敏度高,重复性好,覆盖度高。且TMT标记平行比较多达10个样品!

案例分享

案例一  肿瘤转移: 外核体的特殊靶标或可预测癌症的转移走向

Tumor exosome integrins determine organotropic metastasis. Nature. 2015,  527(7578):329-35.

1,示踪实验证实,具有特定转移倾向的肿瘤细胞来源的外泌体,优先被定居细胞摄取;

2,肿瘤细胞在转移前先释放外泌体,外泌体到达预期转移器官后会被相应的细胞摄取,进而改变这些靶细胞的状态;

3,利用定量蛋白质组学技术,发现外泌体表面的整合素类蛋白质在转移过程中发挥着重要的作用,并决定着靶器官的特异性。如整合素α6 β4 和α6 β1与肺部的转移有关整合素αv β5则与肝脏的转移相关。

案例二  免疫调控: 布氏锥虫通过细胞外囊泡传递毒力因子而引起宿主贫血

Extracellular vesicles from Trypanosoma brucei ediate virulence factor transfer and cause host anemia. Cell. 2016, 164(1-2): 246-257.

1,锥虫的鞭毛膜产生膜纳米管并分解成游离的胞外小泡。致病锥虫通过外泌体传递毒力因子,对人体先天免疫产生抗性;

2,锥虫细胞外囊泡还可以与哺乳动物的红细胞融合,造成红细胞被迅速清除,从而导致贫血。这些数据表明,锥虫细胞外囊泡是一种细胞器,可介导非遗传性毒力因子的转移,使宿主红细胞重构,从而导致贫血。

案例三  植物外泌体: 外泌体蛋白参与植物对病原菌侵染的相应

Extracellular vesicles isolated from the leaf apoplast carry stress-response proteins. Plant Physiol. 2016, 173(1): 728-741.

1,研究者采用不连续对的密度梯度离心从GFP-PEN1转基因植株叶片中分离了胞外囊泡(EVs),利用质谱技术从EVs中鉴定到598个蛋白。将鉴定的蛋白进行基因注释,结果表明有26%的蛋白属于植物应对生物胁迫和非生物胁迫蛋白;

2,拟南芥经假单胞杆菌侵染或用御敌激素水杨酸处理后,植物产生的外泌体数量增加了2倍,表明拟南芥胞外囊泡参与了植物对生物胁迫的响应。

景杰生物通过整合以组学为导向(包括基因蛋白质组学和组蛋白密码组学)的生物标志物发现、以生物标志物为导向的药物研发、以高质量抗体为基础的诊断试剂盒开发这三个环节,逐步构建起“疾病精准分层”、“精准药物研发”、“疾病精准诊断” 三位一体的精准医疗产业化发展的运作链条,从而为精准医疗产业化开创出一片广阔前景, 并开辟出一条可行路径。(生物谷Bioon.com)

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