On-cell screening of mouse hybridomas for improved monoclonal antibody discovery

小鼠杂交瘤的细胞筛选以改进单克隆抗体的发现

• 批准号: 10696698
• 负责人: Rama Devudu Puligedda
• 金额: $ 35万
• 依托单位: OCMS BIO, LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10696698
• 关键词: Address; Advanced Development; Affect; Antibodies; Antibody Specificity; Antibody Therapy; Antigens; B-Lymphocytes; Binding; Biological Assay; Biomedical Research; C-reactive protein; Cell Line; Cell fusion; Cell membrane; Cell secretion; Cell surface; Cells; Clinical; Clinical Research; Clonal Expansion; Clone Cells; Cloning; Comparative Study; Complex; Consumption; Development; Diagnostic; Diagnostics Research; Elements; Engineering; Enzyme-Linked Immunosorbent Assay; Generations; Goals; Human; Human Cloning; Human poliovirus; Hybrid Cells; Hybridomas; Image; Immunize; Immunoglobulin G; Immunoglobulin Genes; Individual; Industrialization; Laboratories; Life; Maintenance; Market Research; Marketing; Methods; Modernization; Monoclonal Antibodies; Mus; N-Methyl-D-Aspartate Receptors; Oryctolagus cuniculus; Phase; Preparation; Process; Property; Rapid screening; Reagent; Reproducibility; Research; Resources; SARS-CoV-2 antigen; Specificity; Splenocyte; Surface; System; Therapeutic; Therapeutic Monoclonal Antibodies; Therapeutic Research; Time; Transgenic Mice; antibody diagnostic; cost; cost effective; fluorescence imaging; human monoclonal antibodies; improved; innovation; novel; process improvement; research and development; screening; success; timeline; tool

SUMMARY OCMS Bio, LLC is seeking to revolutionize monoclonal antibody (mAb) discovery and screening through the development of an on-cell monoclonal antibody (OCMS) screening platform, an innovative cell-based approach to substantially improve the hybridoma method for antibody discovery. Monoclonal antibodies have become essential elements in modern clinical therapeutics, diagnostics, and biomedical research. The hybridoma approach is one of the oldest and most robust methods for generating mAbs. The vast majority of mAbs for clinical therapeutics and diagnostics are generated by hybridoma methods. However, this approach is not without drawbacks, as it is extremely labor intensive and time consuming, because of the need for serial limiting dilutions, clone propagation, and ELISA screening, which substantially limits the total number of mAbs that can be screened. To improve this process, OCMS Bio has developed an approach in which mAbs secreted from OCMS hybridomas are attached to the surface of the cells that secrete them via an innovative anchor-linker system, which enables rapid screening and characterization of expression with standard imaging workflows. This eliminates the need for extensive cloning and ELISA-based screening and is expected to decrease the effort required to screen mAbs by over 80% compared to the traditional hybridoma approach. Furthermore, the OCMS approach supports the discovery of rare mAbs with ideal specificity and developability parameters, which are severely limited in traditional hybridoma approaches due to cloning and screening bottlenecks. OCMS Bio has demonstrated hybridoma formation and the characterization of human mAbs using the OCMS approach in several human systems. OCMS Bio also demonstrated successful hybridoma formation with mouse splenocytes. The goal of this research is to develop a method for using the OCMS approach with mouse splenocytes and rigorously demonstrate 1) reproducible cell fusion for hybridoma formation, 2) stable expression of the OCMS anchor, and 3) that the OCMS approach substantially reduces the time and effort associated with mAb discovery, compared to the traditional hybridoma approach. Success with these goals will have substantial commercial and clinical impact as the OCMS approach will fit within current mAb development workflows and reduce costs, effort, and development timelines for customers to develop novel lifesaving mAb-based therapeutics and diagnostics.

总结 OCMS Bio，LLC正在寻求通过以下方式彻底改变单克隆抗体（mAb）的发现和筛选： 开发细胞上单克隆抗体（OCMS）筛选平台，这是一种基于细胞的创新方法 以实质性地改进用于抗体发现的杂交瘤方法。单克隆抗体已经成为 现代临床治疗学、诊断学和生物医学研究的基本要素。杂交瘤 方法是用于产生mAb的最古老和最稳健的方法之一。绝大多数mAb用于 通过杂交瘤方法产生临床治疗和诊断。不过，这种做法并非没有 缺点是，由于需要连续的有限稀释， 克隆繁殖和ELISA筛选，这基本上限制了可以被克隆的mAb的总数。 筛选出为了改善这一过程，OCMS Bio开发了一种方法，其中从OCMS分泌mAb 杂交瘤通过创新的锚-连接体系统附着于分泌它们的细胞的表面， 其能够用标准成像工作流程快速筛选和表征表达。这 消除了广泛的克隆和基于ELISA的筛选的需要，预计将减少 与传统的杂交瘤方法相比，需要超过80%的筛选mAb。此外，OCMS 该方法支持发现具有理想特异性和可开发性参数的罕见mAb， 由于克隆和筛选瓶颈，在传统杂交瘤方法中受到严重限制。 OCMS Bio已经证明了杂交瘤的形成和使用OCMS的人mAb的表征 在几个人类系统中。OCMS Bio还证明了成功的小鼠杂交瘤形成 脾细胞。本研究的目的是开发一种使用小鼠OCMS方法的方法 并严格证明了1）用于杂交瘤形成可再现的细胞融合，2）稳定的表达 的OCMS锚，以及3）OCMS方法大大减少了与 mAb发现，与传统杂交瘤方法相比。这些目标的成功将产生巨大的影响。 商业和临床影响，因为OCMS方法将适合当前的mAb开发工作流程， 降低成本、工作量和开发时间，帮助客户开发新型救生单克隆抗体 治疗和诊断。