Bundy Bradley C, Franciszkowicz Marc J, Swartz James R
Department of Chemical Engineering, Stanford University, Stanford, California 94305-5025, USA.
Biotechnol Bioeng. 2008 May 1;100(1):28-37. doi: 10.1002/bit.21716.
Virus-like particles (VLP) have received considerable attention for vaccine, drug delivery, gene therapy and material science applications. Although the number of unique VLP and their applications are rapidly growing, the positive impact of VLP applications is limited by the current diverse, expensive, and typically low-yielding production technologies available. These technologies, when scaled, often result in structurally and compositionally inconsistent products. We present Escherichia coli-based cell-free protein synthesis as a production technology to overcome many of the limitations of current VLP production processes. Using this technique, the MS2 bacteriophage coat protein VLP was produced at a yield 14 times the best published production yield. Also, a C-terminally truncated Hepatitis B core protein VLP was produced at similarly high yields (6 x 10(13) VLP/mL). These VLP were found to have comparable characteristics to those produced in vivo. The scalability of this technology was tested without loss in production yields. To our knowledge, this is the first time a prokaryote-based in vitro transcription/translation system has generated a virus-like particle.
病毒样颗粒（VLP）在疫苗、药物递送、基因治疗和材料科学应用方面受到了广泛关注。尽管独特的VLP数量及其应用正在迅速增长，但VLP应用的积极影响受到当前多样化、昂贵且通常产量较低的生产技术的限制。这些技术在扩大规模时，往往会导致产品在结构和组成上不一致。我们提出基于大肠杆菌的无细胞蛋白质合成作为一种生产技术，以克服当前VLP生产过程中的许多限制。使用这种技术，MS2噬菌体衣壳蛋白VLP的产量是已发表的最佳产量的14倍。此外，C末端截短的乙肝核心蛋白VLP也以同样高的产量（6×10¹³ VLP/mL）生产。这些VLP被发现具有与体内产生的VLP相当的特性。该技术的可扩展性在不损失产量的情况下进行了测试。据我们所知，这是首次基于原核生物的体外转录/翻译系统产生病毒样颗粒。
