Tailoring the Hydroxyl Radical Foot-Printing Approach to Provide a Solution for the Higher Order Structural Analysis Needs of the Biopharmaceutical Industry

名为“剪裁羟基自由基”的GenNext阶段II SBIR产品 为高阶结构分析提供解决方案的脚印方法 生物制药业的需求“是对已知的 用于高阶结构分析的新的和改进的工具 生物制药公司。与传统药物不同，生物制药是复杂的， 三维生物分子的非均相混合物，其安全性和有效性是 依赖适当的居屋计划。存在具有不正确高阶结构的蛋白质 (HOS)与严重的药物不良反应有关，这提醒了生物制药公司 业界对居者有其屋计划的关键作用，同时确立需要新的和改善的 Hos Analytics。 一种新兴的HOS分析技术是羟基自由基足迹法(HRPF)。 HRPF涉及通过与羟基反应对蛋白质外部进行不可逆标记 自由基与随后的MS分析以确定蛋白质的外部部分。这个 最广泛使用的产生OH自由基的方法使用快速爆发的紫外光， 并被恰当地称为蛋白质的快速光化学氧化(FPOP)。通常，一个 使用强大而昂贵的紫外光激光器。学术实验室已经证明了 FPOP在HOS分析中的应用；然而，在制药领域的采用一直很少 最好的。我们已经确定了限制#年采用HRPF方法的障碍。 生物制药行业。这些障碍包括：1)使用昂贵的 需要大量安全预防措施的激光；以及2)FPOP的不可复现性 由背景清除OH自由基引起的，这使比较复杂并限制了 学习。因此，目前还没有商业化的FPOP样品制备设备 分析，尽管证明需要居屋的分析能力。 GenNext建议创建了一种改进的执行HRPF分析的方法 通过用闪光氧化系统取代昂贵、危险的激光。此外，一个 将包括定制的内部标准(剂量测定)系统，以便于使用和 提高再现性。这些创新的进步将降低 采用HRPF进行居屋分析，并将加快采用 随之而来的对生物制药研发的影响。除了……之外 仪器销售，GenNext将为希望 在评估福克斯居屋计划对他们计划的影响时，限制前期成本和风险。

The GenNext Phase II SBIR submssion entitled “Tailoring the Hydroxy Radical Foot-Printing Approach to Provide a Solution for the Higher Order Structural Analysis Needs of the Biopharmaceutical Industry” is responsive to the ackowledged need for new and improved tools for higher order structural analysis (HOS) of biopharmaceuticals. Unlike conventional drugs, biopharmaceuticals are complex, heterogeneous mixtures of 3-dimensional biomolecules, whose safety and efficacy is reliant upon proper HOS. The presence of proteins with improper higher order structure (HOS) has been linked to severe adverse drug reactions, alerting the biopharmaceutical industry to the critical role of HOS, while establishing the need for new and improved HOS analytics. An emerging HOS analysis technique is hydroxyl radical foot-printing (HRPF). HRPF involves the irreversible labeling of a protein’s exterior by reaction with hydroxyl radicals with subsequent MS analysis to identify the outer portions of the protein. The most widely used method for generating OH radicals employs a quick burst of UV light, and is appropriately called fast photochemical oxidation of proteins (FPOP). Typically, a powerful and expensive UV laser is used. Academic laboratories have demonstrated the utility of FPOP for HOS analysis; however adoption in pharma has been minuscule at best. We have identified barriers that have limited the adoption of the HRPF approach in the biopharmaceutical industry. These impediments include: 1) The use of expensive lasers that demand substantial safety precautions; and 2) the irreproducibility of FPOP caused by background scavenging of OH radicals that complicate and limit comparative studies. As such, there are no commercial sample preparation devices for FPOP analysis, despite the demonstrated need for the HOS analytical power. The GenNext proposal creates an improved means of performing HRPF analysis by replacing expensive, hazardous lasers with a flash oxidation system. Moreover, a custom internal standard (dosimetry) system will be included to facilitate ease of use and improve reproducibility. These innovative advancements will decrease the barrier to adoption of HRPF for HOS analysis and will result in accelerated adoption with concomitant impact on biopharmaceutical research and development. In addition to instrumentation sales, GenNext will offer a fee-for-service option for its clients who wish to constrain upfront cost and risk, as they evaluate FOX HOS impact to their program.