SBIR TOPIC 269: DEVELOPMENT OF NOVEL PROTEIN EXPRESSION TECHNOLOGIES FOR GLYCOSY

这个项目的目标是开发和验证新的系统，用于高效表达 癌症相关的人类糖基化蛋白。这种系统的可用性将有助于分析癌症相关的 蛋白质，特别是那些低丰度的体液，并加速发展， 有效的癌症诊断和治疗。 细菌和酵母细胞已被广泛用于生产重组人类蛋白质， 低成本，但缺乏类似于翻译后的修改限制了它们的应用。表达 使用哺乳动物细胞通常依赖于瞬时转染或稳定细胞系的建立， 需要高的试剂成本用于大规模生产或长的开发时间。杆状病毒 使用昆虫细胞的表达载体系统（BEVS）以更低的成本和更快的速度提供了一种替代方案 哺乳动物系统的周转时间。在某些情况下，使用昆虫细胞的主要缺点是， 某些修饰如糖基化不同于哺乳动物细胞， 将开发一个完整的系统，在昆虫和哺乳动物细胞中利用BEVS， 纯化方法和生产糖基化蛋白的灵活性。至少有10种这样的蛋白质 在第一阶段生产。这些蛋白质将由MASS进行分析，并提供给更大的科学家。 社区

The objective of this proposed project is to develop and validate novel systems for efficient expression of cancer-related hUman glycosylated proteins. The availability of such systems will aid analysis of cancerrelated proteins, especially those of low abundance from bodily fluids, and accelerate the development of effective cancer diagnostics and therapeutics. Bacteria and yeast cells have been widely used for producing recombinant human proteins with ease and at low costs, but the lack of mammalian-like post-translation modifications limits their applications. Expression using mammalian cells typically relies on either transient transfection or stable cell line establishment, requiring either high reagent costs for large-scale production or long development time. The baculovirus expression vector systems (BEVS) using insect cells provide an alternative at lower cost and faster turnaround time to mammalian systems. The major drawback of using insect cells in some cases is that certain modifications such as glycosylation differ from mammalian cells, A complete system will be developed to utilize BEVS in both insect and mammalian cells with unique purification method and flexibility to produce glycosylated proteins. At least 10 such proteins will be produced in Phase I. These proteins will be analyzed by MASS and made available to the greater scientific community.