Adapting High-Throughput Screening Methods and Assays for Biocontainment Laboratories

Abstract

High-throughput screening (HTS) has been integrated into the drug discovery process, and multiple assay formats have been widely used in many different disease areas but with limited focus on infectious agents. In recent years, there has been an increase in the number of HTS campaigns using infectious wild-type pathogens rather than surrogates or biochemical pathogen-derived targets. Concurrently, enhanced emerging pathogen surveillance and increased human mobility have resulted in an increase in the emergence and dissemination of infectious human pathogens with serious public health, economic, and social implications at global levels. Adapting the HTS drug discovery process to biocontainment laboratories to develop new drugs for these previously uncharacterized and highly pathogenic agents is now feasible, but HTS at higher biosafety levels (BSL) presents a number of unique challenges. HTS has been conducted with multiple bacterial and viral pathogens at both BSL-2 and BSL-3, and pilot screens have recently been extended to BSL-4 environments for both Nipah and Ebola viruses. These recent successful efforts demonstrate that HTS can be safely conducted at the highest levels of biological containment. This review outlines the specific issues that must be considered in the execution of an HTS drug discovery program for high-containment pathogens. We present an overview of the requirements for HTS in high-level biocontainment laboratories.

Laboratorios de Contención: Importancia en la Investigación Biomédica, Enfermedades Emergentes, y la Gestión en Salud Pública

“…Así mismo, se han comenzado a optimizar protocolos para los ensayos de HTS, inicialmente en laboratorios BSL-3, gracias a la simplificación de los equipos necesarios para dichos estudios. Al igual que en el caso de WGS, esto reduce la manipulación de las muestras, favoreciendo la reproducibilidad de los ensayos, lo cual constituye un aspecto crítico para asegurar la confiabilidad de los resultados HTS del laboratorio [40].…”

Development of a Single-Cycle Infectious SARS-CoV-2 Virus Replicon Particle System for Use in Biosafety Level 2 Laboratories

“…Due to the highly contagious nature of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the lack of sufficient immunity in the population, research on SARS-CoV-2 is permitted only under biosafety level 3 (BSL3) containment, which significantly limits SARS-CoV-2 research, i.e., only to institutions with BSL3 infrastructure. In addition, due to physical limitations, high-throughput screening of antiviral drugs can be impractical under BSL3 containment ( 1 , 2 ). Fortunately, research with attenuated or conditional mutants of BSL3/BSL4 pathogens is allowed under lower containment upon demonstration of attenuation and safety ( 3 ).…”

A SARS-CoV-2 nucleocapsid protein TR-FRET assay amenable to high-throughput screening

“…We developed an AlphaLISA assay for the measurement of SARS-CoV‑2 NP as an indicator of virus replication in infected cells 26 . We used the NP as a target viral to develop a high-throughput screening (HTS) assay that could be used in a biosafety level 3 containment facility to identify promising antiviral drugs targeting SARS-CoV-2 multiplication 27 . Targeting the more conserved and highly expressed NP instead of the S would prevent detection failures due to newly emerging SARS-CoV-2 VoC with mutations S mutations and increase sensitivity, respectively.…”