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

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纲要。我们将利用结构和计算生物学的工具来开发探针，以协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元素。此外，这些探针将用于b细胞的选择和分离，以便对其抗体基因位点进行测序，从而能够深入表征分泌抗体。这些能力有望增强我们对感染过程中广泛中和抗体反应如何发展的理解，以及体液免疫系统如何靶向HIV-1 Env糖蛋白的脆弱区域。

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.