High throughput camelid antibody screening as a drug discovery platform

高通量骆驼抗体筛选作为药物发现平台

• 批准号: 9746340
• 负责人: Hiep T Tran
• 金额: $ 17.03万
• 依托单位: ABZYME THERAPEUTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9746340
• 关键词: Affinity; Africa; Agreement; Animal Model; Animals; Antibodies; Antibody Formation; Antibody Therapy; Antigens; Asia; Binding; Biological Assay; Bispecific Antibodies; CD19 gene; CD3 Antigens; California; Catalytic Antibodies; Cell Line; Cities; Clinical Research; Clinical Trials; Collaborations; Communicable Diseases; Contracts; Development; Diagnostic; Disease; Disease Outbreaks; Disease model; ERBB2 gene; Emerging Communicable Diseases; Epidermal Growth Factor Receptor; Epitope Mapping; Escape Mutant; Fees; Funding; Future; Generations; Genes; Goals; Government; Hemagglutinin; Human; Immunization; Immunoglobulin G; In Vitro; Individual; Infection; Influenza; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H5N1 Subtype; Influenza A Virus, H7N9 Subtype; Influenza A virus; Institution; Libraries; Licensing; MDCK cell; Malignant Neoplasms; Monoclonal Antibodies; Mutation; Names; Neuraminidase; Passive Immunization; Pharmacologic Substance; Phase; Plaque Assay; Privatization; Production; Research; Secure; Specificity; Surface; System; T-Lymphocyte; Technology; Testing; Therapeutic; Therapeutic antibodies; Time; Toxin; Universities; Utah; Validation; Viral; Virus; Virus Diseases; Vision; Work; Xenograft procedure; Yeasts; Zoonoses; anti-cancer; anti-influenza; antibody libraries; base; cancer therapy; clinical application; commercial application; commercialization; cost effective; drug discovery; experimental study; human disease; humanized antibody; immunogenic; improved; in vitro Assay; in vivo; infectious disease treatment; influenzavirus; nanobodies; new therapeutic target; novel; novel strategies; payment; programs; scale up; screening; success; therapeutic target; tumor

ABSTRACT. Monoclonal antibodies are used for treatment of a wide range of diseases from cancer to infectious diseases. However, development of antibodies with high affinity and specificity is still time- consuming, labor-intensive and unpredictable, especially to low immunogenic or toxic targets. A new approach for rapid development of antibodies against a variety of diseases, especially against emerging diseases is much needed. Abzyme Therapeutics LLC is actively developing animal-free antibody discovery platforms to accelerate generation of diagnostic and therapeutic antibodies. Phase I objectives have been successfully completed. Specifically, we have created a so-called yeast surface display camelid Self-Diversifying VHH Antibody Library or SDALib as a monoclonal antibody generating system providing a diverse array of antibodies in vitro, without using in vivo immunization. The main feature of the system is the ability to self-generate a library with diversified antibodies by means of hypermutation induction in antibody encoding genes. As proof-of-principle, we have successfully used the platform to isolate a set of single-domain antibodies against N1 neuraminidase (NA) derived from influenza H5N1 virus. Obtained VHH antibodies have been shown to express efficiently in a bacterial system providing an opportunity for accelerated scale-up of antibody production for therapeutic application. Anti-NA VHH antibodies have been purified and characterized in in vitro assays. As proof of principle one (Flu27-8) out of several anti-NA VHH antibodies, that is reactive with N1 NAs of both H5N1 and H1N1 influenza viruses, has been humanized preserving the full anti-NA activity of the parental molecule. Bi- valent anti-NA antibodies that have been produced in a bacterial system as linear fusions showed a significant increase in antibody activity in in vitro assays. Subsequent efforts of Phase II consist of three specific aims that will be performed simultaneously: (i) advance the N1 anti-NA antibodies isolated in Phase I toward clinical application by demonstrating both in vitro and in vivo activity against H5N1 and H1N1 influenza viruses; (ii) expand the application of the platform for developing antibodies targeting both hemagglutinin and NAs of other influenza virus subtypes, e.g. H3N2 and H7N9 which have unusual zoonotic potential and aiming for antibodies cross-reacting with several different neuraminidases (H5N1, H1N1, H3N2 and H7N9); (iii) utilize the antibody discovery platform for developing VHH antibodies against a set of cancer targets followed by production of novel IgG-like VHH-based bispecific humanized antibodies. Phase II efforts are to further extend the application of the technology for rapid development of novel antibodies to therapeutic targets and to advance humanized anti-influenza antibodies isolated in phase I into clinical studies.

摘要。单克隆抗体用于治疗从癌症到癌症的广泛疾病。 传染病然而，开发具有高亲和力和特异性的抗体仍然是时间- 消耗、劳动密集和不可预测，特别是对低免疫原性或毒性靶标。一种新的方法 对于快速开发针对各种疾病，特别是针对新兴疾病的抗体， 非常需要。 Abzyme Therapeutics LLC正在积极开发无动物抗体发现平台，以加速 诊断和治疗抗体的产生。第一阶段的目标已经顺利完成。 具体来说，我们已经创建了一个所谓的酵母表面展示骆驼自我多样化VHH抗体库 或SDALib作为单克隆抗体产生系统，在体外提供多种抗体阵列， 使用体内免疫。该系统的主要特点是能够自行生成一个库， 通过抗体编码基因的超突变诱导使抗体多样化。作为原则证明， 我们已经成功地使用该平台分离出一组抗N1神经氨酸酶的单域抗体 (NA)来源于H5 N1流感病毒。已显示获得的VHH抗体在细胞中有效表达。 细菌系统提供了加速扩大抗体生产的机会， 应用程序.已经在体外测定中纯化和表征了抗NA VHH抗体。就证明了 几种抗NA VHH抗体中的主要一种（Flu 27 -8），其与H5 N1和H5 N2的N1 NA反应， H1N1流感病毒，已经被人源化，保留了亲本分子的全部抗NA活性。双- 已经在细菌系统中作为线性融合物产生的10价抗NA抗体显示出显著的 体外测定中抗体活性增加。 第二阶段的后续工作包括三个同时进行的具体目标： 通过体外和体内试验证明，在I期临床应用中分离的N1抗NA抗体 （ii）扩大该平台的应用范围， 开发靶向血凝素和其他流感病毒亚型（例如H3 N2）的NA的抗体， H7N9具有不寻常的人畜共患病潜力，其目标是与几种不同的抗体交叉反应， 神经氨酸酶（H5 N1、H1N1、H3 N2和H7N9）;（iii）利用抗体发现平台开发 针对一组癌症靶标的VHH抗体，随后产生新的IgG样基于VHH的双特异性抗体。 人源化抗体。第二阶段的工作是进一步扩大该技术的应用， 开发针对治疗靶点的新型抗体并改进人源化抗流感抗体 在I期临床研究中分离。