Design of Antigenically-Specific Probes for Sera Analysis and MAb Isolation

用于血清分析和单克隆抗体分离的抗原特异性探针的设计

• 批准号: 8556160
• 负责人: Peter D Kwong
• 金额: $ 9.9万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8556160
• 关键词: Adsorption; Antibodies; Antibody Formation; Antigens; B-Lymphocytes; Binding; Bioinformatics; Biological; Biological Assay; Computational Biology; Development; Elements; Epitopes; Evaluation; Genes; Goals; HIV-1; Immune system; Individual; Infection; Patients; Play; Process; Proteins; Role; Serologic tests; Serum; Solutions; Specificity; Structure; Surface; Techniques; Therapeutic; Vaccines; Variant; base; computerized tools; design; env Glycoproteins; flexibility; insight; neutralizing antibody; next generation; novel; physical property; research study; response; structural biology; tool

An understanding of the serum responses of both HIV-1-infected individuals and vaccines is necessary for the development of an effective HIV-1 vaccine. Perhaps the most important biological aspect of a serum response is its neutralizing activity, with a close second being a compendium of its antigenic recognition. We will utilize the tools of structural and computational biology to develop probes to assist in the evaluation of the neutralizing activity of sera, and to decipher the HIV-1 elements recognized by both binding and neutralizing antibodies. Further, these probes will be used for the selection and isolation of B-cells so that their antibody gene loci can be sequenced, enabling the in-depth characterization of secreted antibodies. These capabilities are expected to enhance our understanding of how a broadly neutralizing antibody response develops during the course of infection and also of how the humoral immune system targets vulnerable regions on the HIV-1 Env glycoprotein. The tools of computational design allow for the manipulation of both the surface of a protein as well as its interior. Surface manipulation allows for a precise control of antigenicity, whereas interior manipulation allows for physical properties of flexibility and stability to be altered, thereby modulating surface antigencity. Together these tools should allow for a precise understanding of elicited serum responses. Such an understanding should facilitate the iterative structure-based improvement of immunogens. The next-generation sequencing technique, 454 pyrosequencing, allows hundreds of thousands of antibody sequences to be obtained directly from patient sera, thus enabling quantitative analysis of serum response, better understanding of antibody maturation process, and identification of broadly neutralizing antibodies. Due to the natural variation of antibody sequences and inherent sequencing errors, novel bioinformatics techniques have to be developed for antibodyome analysis. These techniques will play a critical role in sera analysis and antibody identification. Identified antibodies can represent natural solutions to antibody optimization, which has potential implications for therapeutic and/or passive transfer studies.

了解HIV-1感染者和疫苗的血清反应对于开发有效的HIV-1疫苗是必要的。也许血清反应最重要的生物学方面是它的中和活性，其次是它的抗原识别概要。我们将利用结构生物学和计算生物学的工具来开发探针，以辅助评估血清的中和活性，并破译结合抗体和中和抗体识别的HIV-1元件。此外，这些探针将用于B细胞的选择和分离，以便对它们的抗体基因位点进行测序，从而能够深入表征分泌的抗体。这些能力有望增强我们对感染过程中如何产生广泛的中和抗体反应，以及体液免疫系统如何针对HIV-1包膜糖蛋白上的脆弱区域的了解。 计算设计的工具允许操控蛋白质的表面和内部。表面操纵允许精确控制抗原性，而内部操纵允许改变柔韧性和稳定性的物理属性，从而调节表面抗原性。这些工具结合在一起，应该可以精确地理解引起的血清反应。这样的理解应该有助于免疫原基于迭代结构的改进。 新一代测序技术454焦磷酸测序技术允许直接从患者血清中获得数十万个抗体序列，从而能够对血清反应进行定量分析，更好地了解抗体成熟过程，并识别广泛的中和抗体。由于抗体序列的自然变异和固有的测序错误，必须开发新的生物信息学技术来进行抗体组分析。这些技术将在血清分析和抗体鉴定中发挥关键作用。识别的抗体可以代表抗体优化的自然解决方案，这对治疗性和/或被动转移研究具有潜在的意义。