Recombinant Hyperimmune Gammaglobulin for Pneumococcal Disease

 描述(由申请人提供)：用于肺炎球菌疾病的重组高免疫丙种球蛋白组织：GigaGen Inc.PI：David S.Johnson，Ph.D.这项SBIR第一阶段项目的具体目标是制造和测试一批用于治疗和预防肺炎球菌感染的天然重组肺炎链球菌免疫球蛋白(RSpIg)，或“超免疫”。体液原发免疫缺陷(PID)是一类以反复感染为特征的疾病，包括常见的可变型免疫缺陷(CVID)和X连锁无丙种球蛋白血症(XLA)。免疫学家使用预防性静脉免疫球蛋白(IVIg)治疗体液性早泄，IVIg是从数千名捐赠者的血清中分离出来的蛋白质池。静脉注射免疫球蛋白可将CVID和XLA患者的肺炎球菌感染减少至少75%(Busse等人，2002年；Bayrakci等人，2005年；Lucas等人，2010年)。据推测，由于IVIg中有0.1%的抗体具有抗肺炎球菌的活性(Mikolajczyk等人，2004年)，许多PID患者需要更高的IVIg剂量来降低肺炎球菌感染率(Orange等人，2010年；Tuerlinck等人，2014年)。靶向肺炎球菌超免疫，即富含抗肺炎球菌抗体的丙种球蛋白，可能具有更高的疗效，而不需要昂贵的剂量。此前，我们开发了GigaLink(Tm)(Johnson等人，2013年)，它使用微流体和多重PCR从抗体库中构建DNA文库，天然的重链和轻链免疫球蛋白切割完好无损。我们将这项技术作为大型制药公司的研究服务进行营销。这一阶段的SBIR将专门改造GigaLink(Tm)，以创造一种重组肺炎球菌超免疫。我们的技术创新是在稳定的CHO表达系统中表达GigaLinkTM天然人类谱系DNA文库。为了制造rSpIg，我们将首先使用GigaLinkTM从最近接种了肺炎病毒(R)23的捐赠者身上捕获B细胞。然后，我们将在CHO中稳定地表达抗体序列，以产生抗肺炎链球菌高免疫蛋白产品。稳定的CHO可以传代并重复使用，以产生数以千计的rSpIg蛋白制剂。由此产生的肺炎球菌超免疫，或rSpIg，将首先使临床创新成为可能，这些创新将帮助免疫缺陷或其他免疫受损的患者。第一阶段将证明我们可以生产一批具有体外活性的rSpIg。在……里面 第二阶段，我们将采取措施建立cGMP生产方案，并对cGMP rSpIg进行毒理学、药代动力学和疗效研究。首先，在慢性和急性情况下，rSpIg将在主要体液缺乏患者中充当传统IVIg的肺炎球菌增强剂。我们还设想儿科医生将使用rSpIg治疗对抗生素无效的儿童的特异性抗体缺陷(SAD)(Sorensen&Moore，2000)。最后，专家告诉我们，我们的技术也将有助于开发病毒的超免疫丙种球蛋白，即用于埃博拉的快速反应。

 DESCRIPTION (provided by applicant): Recombinant Hyperimmune Gammaglobulin for Pneumococcal Disease Organization: GigaGen Inc. PI: David S. Johnson, Ph.D. The Specific Aim of this SBIR Phase I project is to make and test a pilot batch of natural repertoire recombinant Streptococcus pneumonia immunoglobulin (rSpIg), or "hyperimmune", for treatment and prevention of pneumococcal infections. Humoral primary immune deficiency (PID) is a diverse family of disorders, including common variable immune deficiency (CVID) and X-linked agammaglobulinemia (XLA), characterized clinically by recurrent infections. Immunologists treat humoral PID with prophylactic intravenous immunoglobulin (IVIg), which is a pool of proteins isolated from the sera of thousands of donors. IVIg reduces pneumococcal infections in CVID and XLA patients by at least 75% (Busse et al., 2002; Bayrakci et al., 2005; Lucas et al., 2010). Presumably because <0.1% of the antibodies in IVIg have activity against pneumococcus (Mikolajczyk et al., 2004), many PID patients require higher IVIg doses to reduce rates of pneumococcal infections (Orange et al., 2010; Tuerlinckx et al., 2014). A targeted pneumococcal hyperimmune, i.e., a gammaglobulin enriched for anti- pneumococcal antibodies, might have even higher efficacy without requiring costly doses. Previously, we developed GigaLink(tm) (Johnson et al., 2013), which uses microfluidics and multiplexed PCR to build DNA libraries from antibody repertoires, with native heavy and light chain immunoglobulin paring intact. We market the technology as a research service for big pharma. This Phase I SBIR will adapt GigaLink(tm) specifically to create a recombinant pneumococcal hyperimmune. Our technical innovation is to express GigaLinkTM natural human repertoire DNA libraries in a stable CHO expression system. To make rSpIg, we will first use GigaLinkTM to capture B cells from donors recently vaccinated with Pneumovax(r) 23. We will then stably express the antibody sequences en masse in CHO to produce an anti-pneumococcal hyperimmune protein product. The stable CHO can be passaged and used repeatedly to produce thousands of rSpIg protein preps. The resulting pneumococcal hyperimmune, or rSpIg, will first enable clinical innovations that will help patients with immune deficiency or who are otherwise immunocompromised. Phase I will demonstrate that we can produce a test batch of rSpIg that shows in vitro activity. In Phase II, we will take steps to build a cGMP production protocol and perform toxicology, pharmacokinetic, and efficacy studies on cGMP rSpIg. At first, rSpIg will act as a pneumococcal booster for conventional IVIg in primary humoral deficiency patients, both in chronic and acute settings. We also envision that pediatricians would use rSpIg for specific antibody deficiency (SAD) in children who are refractory to antibiotics (Sorensen & Moore, 2000). Finally, experts tell us that our technology will also be useful to develop viral hyperimmune gammaglobulins, i.e., for Ebola rapid response.