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Use of virus vectors for the expression in plants of active full-length and single chain anti-coronavirus antibodies

机译:病毒载体在植物中表达活性全长和单链抗冠状病毒抗体的用途

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To extend the potential of antibodies and their derivatives to provide passive protection against enteric infections when supplied orally in crude plant extracts, we have expressed both a small immune protein (SIP) and a full-length antibody in plants using two different plant virus vectors based on potato virus X (PVX) and cowpea mosaic virus (CPMV). The αSIP molecule consisted of a single chain antibody (scFv) specific for the porcine coronavirus, transmissible gastroenteritis virus (TGEV) linked to the α-CH3 domain from human IgA. To express the full-length IgA, the individual light and heavy chains from the TGEV-specific mAb 6A.C3 were inserted into separate PVX constructs and plants were co-infected with both constructs. Western blot analysis revealed the efficient expression of both the SIP and IgA molecules. Analysis of crude plant extracts revealed that both the plant-expressed αSIP and IgA molecules could bind to and neutralize TGEV in tissue culture, indicating that active molecules were produced. Oral administration of crude extracts from antibody-expressing plant tissue to 2-day-old piglets showed that both the αSIP and full-length IgA molecules can provide in vivo protection against TGEV.
机译:为了扩大抗体及其衍生物在以粗植物提取物口服时提供被动保护以抵抗肠道感染的潜力,我们使用两种不同的植物病毒载体在植物中表达了小免疫蛋白(SIP)和全长抗体感染马铃薯X病毒(PVX)和cow豆花叶病毒(CPMV)。 αSIP分子由对猪冠状病毒,与人IgA的α-CH3结构域相连的可传播性胃肠炎病毒(TGEV)特异的单链抗体(scFv)组成。为了表达全长IgA,将来自TGEV特异性mAb 6A.C3的各个轻链和重链插入单独的PVX构建体中,并用这两种构建体共同感染植物。蛋白质印迹分析揭示了SIP和IgA分子的有效表达。粗植物提取物的分析表明,植物表达的αSIP和IgA分子均可以结合并中和组织培养物中的TGEV,表明产生了活性分子。将来自表达抗体的植物组织的粗提取物口服给药至2天大的仔猪表明,αSIP和全长IgA分子均可在体内提供针对TGEV的保护。

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