Labcode 6B- HIV neutralization Protocol PBMC based HIV-1 Neutralization Assay

This document contains a detailed protocol on detection of neutralizing activity of monoclonal antibodies (mAb) as well as sera/plasma obtained from HIV infected individuals using a PBMC assay. In this assay an extended (24 hr) incubation period of virus and neutralizing agent prior adding the PHA/IL-2 stimulated PBMC is used. Antibody is assessed by measuring the p24 in the culture supernatant using an ‘in-house’ quantitative ELISA.本文件详细阐述了一套实验方案，可通过外周血单个核细胞（PBMC, Peripheral Blood Mononuclear Cell）检测法，对单克隆抗体（mAb）以及HIV感染者血清/血浆的中和活性进行检测。该检测流程中，在添加经植物血凝素（PHA, Phytohemagglutinin）与白细胞介素2（IL-2, Interleukin-2）刺激的外周血单个核细胞前，需将病毒与中和试剂进行24小时的延长孵育。抗体水平通过采用实验室自建（in-house）定量酶联免疫吸附试验（ELISA, Enzyme-Linked Immunosorbent Assay）检测培养上清液中的p24抗原进行评估。