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Development of a surface plasmon resonance assay to measure the binding affinity of wild-type influenza neuraminidase and its H274Y mutant to the antiviral drug zanamivir.
J Mol Recognit. 2015 Feb; 28(2):87-95.JM

Abstract

Influenza is one of the most common infections of the upper respiratory tract. Antiviral drugs that are currently used to treat influenza, such as oseltamivir and zanamivir, are neuraminidase (NA) inhibitors. However, the virus may develop resistance through single-point mutations of NA. Antiviral resistance is currently monitored by a labelled enzymatic assay, which can be inconsistent because of the short half-life of the labelled product and variations in the assay conditions. In this paper, we describe a label-free surface plasmon resonance (SPR) assay for measuring the binding affinity of NA-drug interactions. Wild-type (WT) NA and a histidine 274 tyrosine (H274Y) mutant were expressed in High Five™ (Trichoplusia ni) insect cells. A spacer molecule (1,6-hexanediamine) was site-specifically conjugated to the 7-hydroxyl group of zanamivir, which is not involved in binding to NA, and the construct was immobilized onto a SPR sensor Chip to obtain a final immobilization response of 431 response units. Binding responses obtained for WT and H274Y mutant NAs were fitted to a simple Langmuir 1:1 model with drift to obtain the association (ka) and dissociation (kd) rate constants. The ratio between the binding affinities for the two isoforms was comparable to literature values obtained using labelled enzyme assays. Significant potential exists for an extension of this approach to test for drug resistance of further NA mutants against zanamivir and other antiviral drugs, perhaps paving the way for a reliable SPR biosensor assay that may replace labelled enzymatic assays.

Authors+Show Affiliations

Biomolecular Interaction Centre, University of Canterbury, Private Bag 4800, Christchurch, New Zealand, 8140; Department of Chemical and Process Engineering, University of Canterbury, Private Bag 4800, Christchurch, New Zealand, 8140.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25599664

Citation

Somasundaram, Balaji, et al. "Development of a Surface Plasmon Resonance Assay to Measure the Binding Affinity of Wild-type Influenza Neuraminidase and Its H274Y Mutant to the Antiviral Drug Zanamivir." Journal of Molecular Recognition : JMR, vol. 28, no. 2, 2015, pp. 87-95.
Somasundaram B, Fee CJ, Fredericks R, et al. Development of a surface plasmon resonance assay to measure the binding affinity of wild-type influenza neuraminidase and its H274Y mutant to the antiviral drug zanamivir. J Mol Recognit. 2015;28(2):87-95.
Somasundaram, B., Fee, C. J., Fredericks, R., Watson, A. J., & Fairbanks, A. J. (2015). Development of a surface plasmon resonance assay to measure the binding affinity of wild-type influenza neuraminidase and its H274Y mutant to the antiviral drug zanamivir. Journal of Molecular Recognition : JMR, 28(2), 87-95. https://doi.org/10.1002/jmr.2417
Somasundaram B, et al. Development of a Surface Plasmon Resonance Assay to Measure the Binding Affinity of Wild-type Influenza Neuraminidase and Its H274Y Mutant to the Antiviral Drug Zanamivir. J Mol Recognit. 2015;28(2):87-95. PubMed PMID: 25599664.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of a surface plasmon resonance assay to measure the binding affinity of wild-type influenza neuraminidase and its H274Y mutant to the antiviral drug zanamivir. AU - Somasundaram,Balaji, AU - Fee,Conan J, AU - Fredericks,Rayleen, AU - Watson,Andrew J A, AU - Fairbanks,Antony J, Y1 - 2015/01/20/ PY - 2014/02/28/received PY - 2014/07/02/revised PY - 2014/08/10/accepted PY - 2015/1/21/entrez PY - 2015/1/21/pubmed PY - 2015/9/26/medline KW - antiviral drugs KW - binding kinetics KW - drug resistance KW - influenza neuraminidase KW - surface plasmon resonance KW - zanamivir SP - 87 EP - 95 JF - Journal of molecular recognition : JMR JO - J Mol Recognit VL - 28 IS - 2 N2 - Influenza is one of the most common infections of the upper respiratory tract. Antiviral drugs that are currently used to treat influenza, such as oseltamivir and zanamivir, are neuraminidase (NA) inhibitors. However, the virus may develop resistance through single-point mutations of NA. Antiviral resistance is currently monitored by a labelled enzymatic assay, which can be inconsistent because of the short half-life of the labelled product and variations in the assay conditions. In this paper, we describe a label-free surface plasmon resonance (SPR) assay for measuring the binding affinity of NA-drug interactions. Wild-type (WT) NA and a histidine 274 tyrosine (H274Y) mutant were expressed in High Five™ (Trichoplusia ni) insect cells. A spacer molecule (1,6-hexanediamine) was site-specifically conjugated to the 7-hydroxyl group of zanamivir, which is not involved in binding to NA, and the construct was immobilized onto a SPR sensor Chip to obtain a final immobilization response of 431 response units. Binding responses obtained for WT and H274Y mutant NAs were fitted to a simple Langmuir 1:1 model with drift to obtain the association (ka) and dissociation (kd) rate constants. The ratio between the binding affinities for the two isoforms was comparable to literature values obtained using labelled enzyme assays. Significant potential exists for an extension of this approach to test for drug resistance of further NA mutants against zanamivir and other antiviral drugs, perhaps paving the way for a reliable SPR biosensor assay that may replace labelled enzymatic assays. SN - 1099-1352 UR - https://www.unboundmedicine.com/medline/citation/25599664/Development_of_a_surface_plasmon_resonance_assay_to_measure_the_binding_affinity_of_wild_type_influenza_neuraminidase_and_its_H274Y_mutant_to_the_antiviral_drug_zanamivir_ L2 - https://doi.org/10.1002/jmr.2417 DB - PRIME DP - Unbound Medicine ER -