目前常用的抗体制备技术是杂交瘤技术,但是噬菌体展示技术由于其独特的优势,已有后来者居上的趋势了,那么应该选择哪种技术才是合适的呢?本文将会做一个简单对比介绍。
杂交瘤技术
杂交瘤技术, 它是在体细胞融合技术基础上发展起来的。1975年克勒(Kohler)和米尔斯坦(Milstein)证明, 将骨髓瘤细胞与免疫的动物脾细胞融合,形成能分泌针对该抗原的均质的高特异性的抗体—单克隆抗体。从发明至今,杂交瘤即使早已成为制备单抗的主要技术,现在仍然在免疫组化(immunohistochemistry)和各种不同的细胞生物学技术中发挥着重大的作用。
杂交瘤抗体制备技术流程
许多研究机构也建立杂交瘤核心的实验室来一次性处理多组样品。选择正确的技术取决于你所需的抗体的用途,假如你需要的高产量的抗体,杂交瘤技术或许是正确的选择。但是它的问题在于长时间的制备过程和不完全的抗原表位的识别。
杂交瘤抗体制备优缺点
噬菌体抗体库技术
噬菌体抗体库技术就是将抗体重链可变区和轻链可变区基因与噬菌体的外壳蛋白随机重组,继而感染大肠杆菌,噬菌体感染大肠杆菌后,单链DNA在宿主内复制,并且噬菌体重新组装,分泌到培养基中去,同时不会裂解宿主大肠杆菌。噬菌体与靶标抗原孵育,结合上去的噬菌体分离后,扩增,纯化。这种方法能轻易的筛选大量的克隆。基因工程重组技术的发展带来了噬菌体抗体库技术,尤其在高端抗体应用领域成果丰硕。
噬菌体展示技术简易流程
总的来说,噬菌体抗体库技术在技术上要求比较高。对于技术的掌握和发展都有很高的要求。义翘神州经过近十年的探索和积累,打造出属于自己的噬菌体抗体库技术生产平台,熟练地掌握了基于噬菌体抗体库的鼠单抗、兔单抗的制备技术。它在方法上避开了人工免疫和杂交瘤技术,克服了杂交瘤细胞不稳定的特点。方法简单易行,节约时间,可以通过发酵来大规模生产;筛选容量大,可在数周内筛选100万-1亿个克隆,可获得高亲和力的人源化抗体(在免疫组化实验室效果明显);通过模拟体内免疫过程和天然全套抗体库,可随意克隆任何抗原的抗体。
噬菌体抗体展示技术优缺点
不可否认,噬菌体抗体库技术展示出强劲的发展势头,它的价值也不仅仅局限在抗体制备,它强大的筛选能力已经被应用到免疫学、细胞生物学、药理学、蛋白质工程。当然人们需要进一步解决的问题就是构建库容大、多样性好的抗体库,相信随着此项技术的不断完善,将会有一个广阔的前景。如果你的实验需要利用到抗体,无论是杂交瘤技术还是噬菌体抗体库技术来制备的抗体,都有各自的一些长处和局限,希望以上的介绍能给您的选择做出一丁点的参考。
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