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On-chip manufacturing of synthetic proteins for point-of-care therapeutics

机译:用于护理点治疗剂的合成蛋白片的片上制造

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Therapeutic proteins have recently received increasing attention because of their clinical potential. Currently, most therapeutic proteins are produced on a large scale using various cell culture systems. However, storing and transporting these therapeutic proteins at low temperatures makes their distribution expensive and problematic, especially for applications in remote locations. To this end, an emerging solution is to use point-of-care technologies that enable immediate and accessible protein production at or near the patients bedside. Here we present the development of Therapeutics-On-a-Chip (TOC), an integrated microfluidic platform that enables point-of-care synthesis and purification of therapeutic proteins. We used fresh and lyophilized materials for cell-free synthesis of therapeutic proteins on microfluidic chips and applied immunoprecipitation for highly efficient, on-chip protein purification. We first demonstrated this approach by expressing and purifying a reporter protein, green fluorescent protein. Next, we used TOC to produce cecropin B, an antimicrobial peptide that is widely used to control biofilm-associated diseases. We successfully synthesized and purified cecropin B at 63ng/l within 6h with a 92% purity, followed by confirming its antimicrobial functionality using a growth inhibition assay. Our TOC technology provides a new platform for point-of-care production of therapeutic proteins at a clinically relevant quantity.
机译:由于它们的临床潜力,疗效蛋白最近得到了越来越长的关注。目前,大多数治疗蛋白质使用各种细胞培养系统在大规模上产生。然而,在低温下储存和运输这些治疗性蛋白质使其分配昂贵和有问题,特别是对于远程位置的应用。为此,新兴解决方案是使用护理点技术,可在患者床边或附近使用立即和可访问的蛋白质生产。在这里,我们介绍了芯片治疗剂(TOC),一种集成的微流体平台,可实现治疗蛋白质的护理点合成和纯化。我们使用新鲜和冻干材料,用于无细胞合成治疗蛋白质的微流体芯片,并应用免疫沉淀,用于高效,片上蛋白质纯化。我们首先通过表达和纯化报告蛋白,绿色荧光蛋白来证明这种方法。接下来,我们使用TOC生产Cecropin B,一种广泛用于控制生物膜相关疾病的抗微生物肽。我们在6小时内成功合成并纯化了6小时内的Cecropinb,纯度为92%,然后使用生长抑制测定来确认其抗微生物功能。我们的TOC技术为临床相关数量提供了治疗蛋白的护理点生产的新平台。

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