High throughput HIV-1 microneutralization assay

Abstract

Antibodies directed against the HIV-1 Envelope glycoprotein are routinely assessed for in vitro viral neutralization. The most commonly used assay is based on infection of a cell line that expresses luciferase upon infection by HIV-1. Recently, several groups have published techniques for single-cell culture of B cells in 384-well plates. The standard 96 well neutralization assay format is not suitable for screening the large number of low-volume samples produced in such cultures. Therefore, we adapted the assays to a "microneutralization" format, which allows the use of 20 ul of undiluted antibody sample and permits an assessment of the relative potency of each sample. The method can be adapted for assays by hand or using robotic pipetting devices.