Bioprocessing of Plasma Therapeutic Proteins using Sequential Affinity Monolithic

使用顺序亲和整体法对血浆治疗蛋白进行生物处理

• 批准号: 7272450
• 负责人: YOW-PIN LIM
• 金额: $ 17.05万
• 依托单位: PROTHERA BIOLOGICS, LLC
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7272450
• 关键词: Accounting; Address; Affinity; Albumins; Anticoagulants; Binding; Biological Products; Biological Response Modifier Therapy; Blood; Blood Coagulation Factor; Businesses; Class; Clinical; Collection; Complex; Condition; Coupled; Development; Disease; Dyes; Ethanol; Evaluation; Explosion; Extravasation; Fractionation; Future; Generations; Genetic Engineering; Goals; Human; Immobilization; Immunoglobulin Fragments; Immunoglobulin G; Immunoglobulins; Industry; Inflammation; Intravenous Immunoglobulins; Investigation; Laboratories; Ligands; Link; Marketing; Methacrylates; Methods; Numbers; Patients; Peptide antibodies; Peptides; Performance; Pharmaceutical Preparations; Phase; Plasma; Plasma Proteins; Play; Preparation; Prevalence; Process; Production; Protease Inhibitor; Proteins; Protocols documentation; Recombinants; Research; Role; Safety; Science; Secure; Sepsis; Serine Proteinase Inhibitors; Shapes; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Source; Speed; Standards of Weights and Measures; Structure; Technology; Testing; Therapeutic; Therapeutic Uses; Today; Tube; United States; United States Food and Drug Administration; Validation; Vertebral column; World War II; alpha 1-Antitrypsin; base; bioprocess; concept; cost; density; design; expectation; improved; inhibitor/antagonist; innovation; inter-alpha-inhibitor; novel; novel therapeutics; novel virus; optimism; scale up; size; therapeutic protein

DESCRIPTION (provided by applicant): Plasma fractionation is one of the largest industry segments in global therapeutic protein manufacture. The United States plays an important role in this industry since it represents the largest plasma collection and manufacturing base that accounts for more than 50% of the world supply of plasma. Even though more than 22 million liters of plasma are collected each year for fractionation, this amount cannot meet the high demand for plasma derivatives worldwide such as albumin (about 500 tons), FVIII (2 kg) and IgG (40 tons). Consequently, optimal fractionation methods are important to increase the various therapeutic proteins extracted from plasma and to improve the yields of the purified products. In spite of the continued prevalence of protein based blood products in the clinical arena, the methods currently used to obtain them known as fractionation, are outdated and inefficient, a case in point being ethanol fractionation, a purification process developed during World War II that still remains the backbone of most fractionation strategies in use today The recent addition of chromatographic methods for industrial scale plasma fractionation has become a standard approach in the last few years that has played a central role in the development of a new generation of highly purified therapeutic plasma derivatives that includes coagulation factors, protease inhibitors and anticoagulants. Nevertheless, the scarcity of human plasma has created a pressing need for improving the yield of purified plasma proteins obtained using current fractionation technologies. The FDA through the Center of Biologics Evaluation (CDER) has long recognized that the development of biological products is becoming increasingly challenging. The agency has suggested that superior product development sciences are needed to address the challenges of greater efficiency and lower costs, essential goals if the United States is going to remain a leader in the develop of plasma derived therapeutics. In recognition of this need, in this proposal, we will develop a strategy to radically improve the bioprocessing of plasma derived protein therapeutics by using specific affinity ligands immobilized on a new generation of chromatographic supports, an approach that should increase the yield, speed of production and quality of plasma derived products. Specifically, we will make use of Convective Interaction Media (CIM), a recently developed revolutionary chromatographic support based on methacrylate monoliths. In the proposed studies, a sequential chromatographic separation of human plasma on several affinity CIM supports will be designed to streamline the fractionation process of existing plasma derivatives such as immunoglobulins and clotting factors but also of newer classes of plasma derived products such as protease inhibitors. In Specific Aim 1, we will develop affinity supports specific for various plasma proteins by immobilizing antibodies, peptides or synthetic dyes on epoxy-activated CIM columns. A major goal will be to optimize the immobilization conditions by assessing performance, stability, and reusability of ligands coupled under different conditions and densities. In Specific Aim 2, we will develop purification protocols based on serially linked-affinity CIM discs with the goal of obtaining a high level of purity and activity in a single step purification. We will also evaluate the yield and purity following scale-up of CIM separation protocols, a necessary step towards industrial level production that will be facilitated by the identical performance and purification profiles of short monoliths independently from their size or shape (disk, column or tube), The results of these investigations should provide the "proof of concept" for the use of CIM technology in industrial fractionation of human plasma.

描述（由申请人提供）：血浆分馏是全球治疗蛋白制造中最大的行业领域之一。美国在该行业中起着重要的作用，因为它代表了最大的等离子体收集和制造基地，占全球血浆供应的50％以上。即使每年收集超过2200万升的等离子体来分馏，但该数量无法满足全球对等离子体衍生物的高需求，例如白蛋白（约500吨），FVIII（2 kg）和IgG（40吨）。因此，最佳分级方法对于增加从血浆中提取的各种治疗蛋白并提高纯化产物的产量很重要。 In spite of the continued prevalence of protein based blood products in the clinical arena, the methods currently used to obtain them known as fractionation, are outdated and inefficient, a case in point being ethanol fractionation, a purification process developed during World War II that still remains the backbone of most fractionation strategies in use today The recent addition of chromatographic methods for industrial scale plasma fractionation has become a standard approach in the last few years that has played a central role在开发新一代高度纯化的治疗性血浆衍生物中，包括凝结因子，蛋白酶抑制剂和抗凝剂。然而，人类血浆的稀缺性迫切需要提高使用当前分级技术获得的纯化等离子体蛋白的产量。通过生物制剂评估中心（CDER）的FDA长期以来已经认识到，生物产品的发展变得越来越具有挑战性。该机构建议，需要卓越的产品开发科学来应对更高效率和降低成本的挑战，如果美国将继续成为血浆衍生的治疗剂的领导者，那么基本的目标。为了认识到这一需求，在本提案中，我们将通过使用固定在新一代色谱支持的特定亲和力配体来从根本上改善血浆衍生蛋白质疗法的生物处理，这种方法应提高血浆衍生产品的产量，生产速度和质量。具体而言，我们将利用对流相互作用介质（CIM），这是一种基于甲基丙烯酸酯整体的最近开发的革命性色谱支持。在拟议的研究中，将旨在简化现有血浆衍生物的分馏过程，例如免疫球蛋白和凝血因子，以及较新的等离子体衍生产物（如蛋白酶抑制剂），将人血浆在几个亲和力CIM支持上进行顺序色谱分离。在特定的目标1中，我们将通过将抗体，肽或合成染料固定在环氧激活的CIM柱上来开发对各种血浆蛋白的特异性支持。一个主要目标是通过评估在不同条件和密度下耦合的配体的性能，稳定性和可重复使用性来优化固定条件。在特定的目标2中，我们将基于串行连接的亲密CIM盘制定纯化协议，其目的是在单步纯化中获得高水平的纯度和活动。我们还将在CIM分离协议扩大规模后评估产量和纯度，这是迈向工业水平生产的必要步骤，这将由短量的相同性能和纯化概况促进与其大小或形状（磁盘，柱子或管）独立的相同性能和净化概况，这些研究的结果应为CIM技术在工业plasmma of Human Plasma of Hulton Plasma of Human Plasma of Munthrial Plasma of Munthlial Plasmammma of Concept of Coock of Supple。