Ultrasensitive Env Detection Assay for Broadly Neutralizing Antibody Screening

用于广泛中和抗体筛选的超灵敏包膜检测分析

基本信息

批准号：
10676393
负责人：
Alberto Bosque
金额：
$ 45.02万
依托单位：
GEORGE WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-12 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10676393
关键词：
Address Aliquot Antibodies Binding Biological Biological Assay CLIA certified Cells Clinical Trials Cloning Combined Modality Therapy Consumption Data Detection Documentation Enrollment Epitopes Equipment HIV HIV Antibodies HIV-1 Hour Immunotherapy In Vitro Individual Infusion procedures Length Measurement Measures Methodology Methods Minority Nature Participant Performance Peripheral Blood Mononuclear Cell Persons Phase Phenotype Plasma Proviruses Publishing Qualifying Quality Control Reagent Reporting Resistance Sampling Signal Transduction Specific qualifier value System Technology Testing Therapeutic Time Variant Viral Virion Virus Virus Replication antibody detection clinical development clinical efficacy clinical predictors clinical trial participant clinically relevant detection assay detector env Gene Products env Genes gag Gene Products improved multidisciplinary neutralizing antibody novel resistant strain screening treatment strategy

项目摘要

PROJECT SUMMARY Broadly neutralizing antibodies (bNAbs) are a promising immunotherapy that can be incorporated in many strategies for the treatment and/or cure of HIV-1. There is a clear association between the neutralization sensitivity of virus and how effective bNAbs are in suppressing virus replication during clinical trials, but the precise nature of this relationship is still not clear. Individual clinical trials have reached different conclusions about the utility of pre-screening participants’ virus for in vitro neutralization sensitivity before enrollment into clinical trials mainly due to two obstacles: length of assay time and difficulty in obtaining the correct samples to test. The Bosque lab has recently published an ultrasensitive method to detect p24 gag protein down to the fg/ml level, and the assay was validated in ex vivo cells from PLWH. Here we will apply this novel methodology to address these 2 obstacles by developing an assay that gives an indirect readout of neutralization sensitivity but in a very short amount of time and directly in cell lysates. We will achieve this ultra-sensitive Env binding assay by developing it in three parts for the first R61 phase: Aim 1 will optimize the capture and detector antibodies by testing a matrix of bNAbs against diverse pseudoviruses with known neutralization sensitivities, Aim 2 will optimize biological matrices by testing Env detection in plasma and cell lysates, and Aim 3 will determine assay precision for ratios of sensitive and resistant virus in a diverse viral quasispecies. We will validate and qualify this ultra-sensitive Env binding assay during the second R33 phase in two parts: Aim 4 will validate the assay through post-hoc testing of well-studied clinical trial samples. Aim 5 will implement correct quality systems for this assay to prepare for CLIA qualification. Our multi-disciplinary team has all the expertise necessary to achieve these aims that, when completed, result in an ultra-sensitive binding assay for Env protein to screen clinical trial participants that is time-efficient, accurate and qualified.

项目摘要广泛中和抗体（BNAB）是一种承诺免疫疗法，可以纳入许多 HIV-1治疗和/或治疗的策略。中和之间有明显的关联病毒的敏感性以及BNAB在临床试验中抑制病毒复制方面的有效性，但是这种关系的确切性质仍然不清楚。个别临床试验得出了不同的结论关于在入学之前，筛查参与者病毒在体外神经敏感性之前的实用性临床试验主要是由于两个障碍：分析时间的长度和难以获得正确样本测试。 Bosque Lab最近发表了一种超敏化方法，可检测到FG/ML的P24 GAG蛋白水平，该测定在PLWH的离体细胞中验证。在这里，我们将把这种新颖的方法应用于通过开发一个间接读数神经抑制的测定法，解决这两个障碍，但在很短的时间内，直接在细胞裂解物中。我们将实现这种超敏感的ENV结合测定通过在第一个R61阶段以三个部分开发它：AIM 1将通过测试具有已知Neurostimuli的潜水假病毒的BNAB矩阵，AIM 2 Will 通过测试血浆和细胞裂解物中的ENV检测来优化生物材料，AIM 3将确定测定法敏感和抗性病毒比率的精度。我们将验证和资格在第二个R33期间，这种超敏感的ENV结合分析分为两个部分：AIM 4将验证该测定法通过事后测试进行良好的临床试验样品。 AIM 5将实施正确的质量系统这项评估为CLIA资格做准备。我们的多学科团队拥有实现所需的所有专业知识这些目的是，完成后，为筛查临床试验提供了一种超敏感的结合测定时间效率，准确和合格的参与者。