Post by Nadica (She/Her) on Oct 13, 2024 23:56:02 GMT
Functional Activity and Binding Specificity of small Ankyron Repeat Proteins against SARS-CoV-2 variants - Preprint posted Oct 11, 2024
Abstract
Effective management of COVID-19 requires clinical tools to treat the disease in addition to preventive vaccines. Several recombinant mAbs and their cocktails have been developed to treat COVID-19 but these have limitations. Here, we evaluate Ankyrons that were generated to bind with high affinity to the SARS-CoV-2 virus. Ankyrons are ankyrin repeat proteins comprised of repetitions a structural module. Each module consists of a beta-turn followed by two antiparallel alpha-helices. The Ankyrons (TM) can be generated synthetically and like antibodies can bind with high affinity to almost any target. We assessed Ankyrons that were generated against the wild-type SARS-CoV-2 and the Delta and Omicron variants in a binding assay. We determined that all Ankyrons were specific in that they did not bind to MERS, a related coronavirus. While all Ankyrons bound with high affinity to the variant they were generated against, some also showed cross-reactivity to all three SARS-CoV-2 variants. Binding assays are useful for screening analytes but do not provide information about clinical effectiveness. Therefore, we used a pseudoviruses based neutralization assay to show that five of the Ankyrons evaluated neutralized all three strains of SARS-CoV-2. We have provided a workflow for the generation and evaluation of Ankyrons against a viral target. Such a workflow could provide the basis for a platform technology to rapidly developing a new class of therapeutic drugs to rapidly combat diverse pathogens when they emerge.
Abstract
Effective management of COVID-19 requires clinical tools to treat the disease in addition to preventive vaccines. Several recombinant mAbs and their cocktails have been developed to treat COVID-19 but these have limitations. Here, we evaluate Ankyrons that were generated to bind with high affinity to the SARS-CoV-2 virus. Ankyrons are ankyrin repeat proteins comprised of repetitions a structural module. Each module consists of a beta-turn followed by two antiparallel alpha-helices. The Ankyrons (TM) can be generated synthetically and like antibodies can bind with high affinity to almost any target. We assessed Ankyrons that were generated against the wild-type SARS-CoV-2 and the Delta and Omicron variants in a binding assay. We determined that all Ankyrons were specific in that they did not bind to MERS, a related coronavirus. While all Ankyrons bound with high affinity to the variant they were generated against, some also showed cross-reactivity to all three SARS-CoV-2 variants. Binding assays are useful for screening analytes but do not provide information about clinical effectiveness. Therefore, we used a pseudoviruses based neutralization assay to show that five of the Ankyrons evaluated neutralized all three strains of SARS-CoV-2. We have provided a workflow for the generation and evaluation of Ankyrons against a viral target. Such a workflow could provide the basis for a platform technology to rapidly developing a new class of therapeutic drugs to rapidly combat diverse pathogens when they emerge.