Highly efficient transient expression of functional recombinant antibodies in lettuce
Highly efficient transient expression of functional recombinant antibodies in lettuce
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DOI：
10.1016/j.plantsci.2005.03.031
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发表时间：
2005-08-01
期刊：
影响因子：
5.2
通讯作者：
Wong, HC
中科院分区：
文献类型：
--
作者：
Negrouk, V;Eisner, G;Wong, HC
一种高效、可扩展的瞬时表达系统已被开发出来，并用于在生菜中生产重组抗体。该瞬时表达系统以生菜(Lactuca sativa L.)真空渗入携带抗体基因的根癌农杆菌，克隆到一个或两个单独的表达载体中。生菜可以从商业上获得(杂货店或直接从种植者那里获得)，因此不需要植物生长设施。与之前描述的系统相比，这种瞬时表达系统被广泛修改以降低成本和操作，并提供显著提高的抗体产生水平。优化的农业渗滤程序在不到一周的时间内，每公斤生菜叶片组织产生高达20-80 mg的功能抗体。该方法已被用于生产人源化的IgG1kappa抗组织因子抗体(HoAT)以及其他抗体。结果表明，在生菜中瞬时表达重组抗体是一种非常高效、廉价且易于扩展的方法。因此，它是小规模生产功能活性抗体和其他药用重要蛋白质的有用工具，可用于促进抗体的人源化或最优化，或生产足以进行体内研究的抗体(范围在100毫克范围内)。(C)2005爱思唯尔爱尔兰有限公司。保留所有权利。
An efficient, scalable transient expression system has been developed and characterized for production of recombinant antibodies in lettuce. The transient expression system utilizes lettuce (Lactuca sativa L.) vacuum-infiltrated with Agrobacterium tumefaciens bearing antibody genes cloned into a single or two separate expression vectors. The lettuce can be obtained commercially (grocery stores or directly from growers), so plant growth facilities are not required. Compared to previously described systems, this transient expression system is extensively modified to reduce cost and manipulations and provides for significantly increased levels of antibody production. The optimized agro-infiltration procedure produced as much as 20-80 mg of functional antibody per kilogram of fresh lettuce leaf tissue in less than a week. This procedure has been used to produce a humanized IgG1 kappa anti-tissue factor antibody (hOAT) as well as other antibodies. The results demonstrate that transient expression of recombinant antibodies in lettuce is very efficient, inexpensive and readily scalable. Thus, it is a useful tool for small-scale production of functionally active antibodies and other pharmaceutically important proteins and can be used to facilitate humanization or optimization of antibodies or to produce quantities of antibody (in the range of 100 mg) sufficient for in vivo studies. (c) 2005 Elsevier Ireland Ltd. All rights reserved.