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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的。单克隆抗体用于治疗从癌症到多种疾病传染病。然而，开发具有高亲和力和特异性的抗体仍然需要时间。消耗性、劳动密集型和不可预测性，特别是对于低免疫原性或毒性目标。一种新方法快速开发针对多种疾病，特别是针对新出现疾病的抗体非常需要。 Abzyme Therapeutics LLC 正在积极开发非动物抗体发现平台，以加速诊断和治疗抗体的产生。第一阶段目标已顺利完成。具体来说，我们创建了一个所谓的酵母表面展示骆驼科自我多样化 VHH 抗体库或 SDALib 作为单克隆抗体生成系统，在体外提供多种抗体，无需使用体内免疫。该系统的主要特点是能够自行生成库通过抗体编码基因的超突变诱导使抗体多样化。作为原理证明，我们已成功利用该平台分离出一组针对 N1 神经氨酸酶的单域抗体 (NA) 源自流感 H5N1 病毒。获得的 VHH 抗体已被证明可以在细菌系统为加速扩大治疗用抗体生产提供了机会应用。抗 NA VHH 抗体已在体外测定中纯化和表征。作为证明几种抗 NA VHH 抗体中的原理之一（Flu27-8），与 H5N1 和 H5N1 的 N1 NA 发生反应 H1N1 流感病毒已人源化，保留了亲本分子的全部抗 NA 活性。双- 在细菌系统中作为线性融合产生的价抗 NA 抗体显示出显着的体外测定中抗体活性增加。第二阶段的后续工作包括将同时实现的三个具体目标：（i）推进通过体外和体内验证，在 I 期中分离的 N1 抗 NA 抗体走向临床应用针对 H5N1 和 H1N1 流感病毒的体内活性；（二）拓展平台应用范围开发针对其他流感病毒亚型（例如流感病毒亚型）的血凝素和 NA 的抗体H3N2 和 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发生交叉反应神经氨酸酶（H5N1、H1N1、H3N2 和 H7N9）； (iii) 利用抗体发现平台进行开发针对一组癌症靶点的 VHH 抗体，然后生产基于 VHH 的新型 IgG 样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，以实现快速开发针对治疗靶点的新型抗体并推进人源化抗流感抗体在I期临床研究中分离。