McLean Michael D, Almquist Kurt C, Niu Yongfing, Kimmel Rhonda, Lai Zengzu, Schreiber John R, Hall J Christopher
Department of Environmental Biology, University of Guelph, Guelph, Ontario, N1G 2W1 Canada.
Antimicrob Agents Chemother. 2007 Sep;51(9):3322-8. doi: 10.1128/AAC.00366-07. Epub 2007 Jul 2.
The production of a recombinant human IgG1 in transgenic tobacco was examined to determine whether a plant-derived antibody could recruit immune system effector function against a bacterial pathogen. A plant transformation vector was engineered to contain genes for a human kappa light chain and a human gamma-1 heavy chain with V(H) and V(L) sequences from a previously identified human IgG2 monoclonal antibody (MAb) that specifically binds to and opsonizes Pseudomonas aeruginosa serotype O6ad. Unique NcoI and NotI restriction sites were incorporated to flank these variable sequences, resulting in a plant transformation vector that could be engineered for expression of any other human IgG1 antibody, requiring only the substitution of other V(H) and V(L) antigen-binding coding sequences. The plant-produced IgG1 was determined to have high-mannose glycan content and to be capable of mediating opsonophagocytosis of P. aeruginosa serotype O6ad in vitro using human complement and human polymorphonuclear leukocytes. Thus, MAbs produced in plants from this vector could provide human IgG1 MAbs for targeting other pathogens that require the recruitment of immune system effector functions.
研究了转基因烟草中重组人IgG1的产生,以确定植物源抗体是否能募集免疫系统效应功能来对抗细菌病原体。构建了一个植物转化载体,其中包含人κ轻链和人γ-1重链的基因,其V(H)和V(L)序列来自先前鉴定的特异性结合并调理铜绿假单胞菌血清型O6ad的人IgG2单克隆抗体(MAb)。引入了独特的NcoI和NotI限制性酶切位点位于这些可变序列两侧,从而得到一个植物转化载体,该载体可被设计用于表达任何其他人类IgG1抗体,只需替换其他V(H)和V(L)抗原结合编码序列即可。经测定,植物产生的IgG1具有高甘露糖聚糖含量,并且能够在体外使用人补体和人多形核白细胞介导铜绿假单胞菌血清型O6ad的调理吞噬作用。因此,从该载体在植物中产生的单克隆抗体可为靶向其他需要募集免疫系统效应功能的病原体提供人IgG1单克隆抗体。