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.

对HIV-1感染个体和疫苗的血清反应的理解对于开发有效的HIV-1疫苗是必要的。血清反应中最重要的生物学方面也许是其中和活性，第二个是其抗原识别的纲要。我们将利用结构和计算生物学的工具来开发探针，以帮助评估血清中和活性，并破译通过结合和中和抗体识别的HIV-1元素。此外，这些探针将用于选择和分离B细胞，以便可以对其抗体基因位点进行测序，从而实现分泌抗体的深入表征。预计这些能力将增强我们对在感染过程中广泛中和抗体反应的理解，以及体液免疫系统如何靶向HIV-1 ENV糖蛋白的脆弱区域。 计算设计的工具允许操纵蛋白质的表面及其内部。表面操作可以精确控制抗原性，而内部操作则可以改变灵活性和稳定性的物理特性，从而调节表面抗活性。这些工具共同使对引起的血清反应有准确的理解。这样的理解应促进免疫原料的基于迭代结构的改进。 下一代测序技术，454焦磷酸测序，可以直接从患者血清中获得数十万个抗体序列，从而实现了血清反应的定量分析，更好地了解抗体成熟过程，并鉴定出广泛中和中和抗体的鉴定。由于抗体序列和固有测序误差的自然变化，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识别中起关键作用。鉴定的抗体可以代表抗体优化的自然解决方案，该抗体对治疗和/或被动转移研究具有潜在的影响。