Effect of thiomersal on dissociation of intact (146S) foot-and-mouth disease virions into 12S particles as assessed by novel ELISAs specific for either 146S or 12S particles

Harmsen, Michiel M.; Fijten, H.P.D.; Westra, Douwe F.; Coco-Martin, J.M.

doi:10.1016/j.vaccine.2011.01.069

Cited by 28 publications

(29 citation statements)

References 33 publications

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“…The radiolabeled virus-based method we had used so far to quantify stability presents technical inconveniences when many mutants need to be tested. Thus, we have used instead an immunochemical method based on an engineered antibody, VHH2 M3ggsVI-4 Q6E (Harmsen et al, 2008), for the specific quantification of pentameric subunits produced on dissociation of FMDV particles (Harmsen et al, 2011). We have compared this method with the radioactivitybased procedure.…”

Section: Effects On Fmdv Infectivity and Thermostability Of Capsid Mumentioning

confidence: 99%

“…This assay is based on the specific detection of free FMDV pentamers in ELISA (Harmsen et al, 2011) by a genetic fusion of two single-domain antibody fragments (VHH2) termed M3ggsVI-4 Q6E (Harmsen et al, 2008). One of the VHHs, M3, binds FMDV pentamers, whereas the second VHH, VI-4 Q6E , binds to porcine immunoglobulin (this latter VHH has no function in this assay).…”

Section: Thermal Dissociation Assays Using Immunodetectionmentioning

confidence: 99%

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Identification of the Structural Basis of Thermal Lability of a Virus Provides a Rationale for Improved Vaccines

et al. 2014

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Virus stability and dynamics play critical roles during infection. Some viruses, including foot-and-mouth disease virus (FMDV), are surprisingly prone to thermal dissociation outside the cell. The structural bases and functional implications of this distinctive trait were essentially unknown. This study (1) uncovers the structural determinants of FMDV thermolability, (2) investigates the relationship between virus thermolability and infectivity, and (3) provides a structure-based rationale for engineering thermostable virus particles to develop improved vaccines and nanocontainers. The results reveal that negatively charged residues close to protein-protein interfaces exert electrostatic repulsions between capsid subunits and mediate the sensitivity of the virion to thermal dissociation, even at neutral pH. Based on these results, a series of fully infectious virions of increased thermostability were engineered by individually removing different carboxylates involved in intersubunit repulsions. The implications for virus biology and the design of thermostable vaccines are discussed.

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Section: Effects On Fmdv Infectivity and Thermostability Of Capsid Mumentioning

confidence: 99%

Section: Thermal Dissociation Assays Using Immunodetectionmentioning

confidence: 99%

Identification of the Structural Basis of Thermal Lability of a Virus Provides a Rationale for Improved Vaccines

et al. 2014

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“…They were suitable for detection of A or O serotype strains ( 15 , 16 ). We have recently developed two DAS ELISAs using two recombinant llama single-domain antibody fragments (VHHs) that bind specifically to either 146S particles or 12S particles of strain O 1 Manisa ( 17 , 18 ). In each ELISA, the same VHH was used for coating as well as for detection of captured antigen using biotinylated VHH.…”

Section: Introductionmentioning

confidence: 99%

“…The DAS ELISA employing 12S-specific VHH M3 could detect FMDV antigen of several A, O, and Asia 1 strains but not SAT2 strain. We were also able to measure 146S particles of A and Asia 1 serotype strains employing the M3 DAS ELISA of heated and untreated samples ( 17 ). However, this latter ELISA approach is not suitable for detecting 146S particles in the presence of higher concentrations of 12S particles.…”

Section: Introductionmentioning

confidence: 99%

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Isolation of Single-Domain Antibody Fragments That Preferentially Detect Intact (146S) Particles of Foot-and-Mouth Disease Virus for Use in Vaccine Quality Control

et al. 2017

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Intact (146S) foot-and-mouth disease virus (FMDVs) can dissociate into specific (12S) viral capsid degradation products. FMD vaccines normally consist of inactivated virions. Vaccine quality is dependent on 146S virus particles rather than 12S particles. We earlier isolated two llama single-domain antibody fragments (VHHs) that specifically recognize 146S particles of FMDV strain O1 Manisa and shown their potential use in quality control of FMD vaccines during manufacturing. These 146S-specific VHHs were specific for particular O serotype strains and did not bind strains from other FMDV serotypes. Here, we describe the isolation of 146S-specific VHHs against FMDV SAT2 and Asia 1 strains by phage display selection from llama immune libraries. VHHs that bind both 12S and 146S particles were readily isolated but VHHs that bind specifically to 146S particles could only be isolated by phage display selection using prior depletion for 12S particles. We obtained one 146S-specific VHH—M332F—that binds to strain Asia 1 Shamir and several VHHs that preferentially bind 146S particles of SAT2 strain SAU/2/00, from which we selected VHH M379F for further characterization. Both M332F and M379F did not bind FMDV strains from other serotypes. In a sandwich enzyme-linked immunosorbent assay (ELISA) employing unlabeled and biotinylated versions of the same VHH M332F showed high specificity for 146S particles but M379F showed lower 146S-specificity with some cross-reaction with 12S particles. These ELISAs could detect 146S particle concentrations as low as 2.3–4.6 µg/l. They can be used for FMD vaccine quality control and research and development, for example, to identify virion stabilizing excipients.

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Structural and molecular basis for foot-and-mouth disease virus neutralization by two potent protective antibodies

et al. 2021

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Effect of thiomersal on dissociation of intact (146S) foot-and-mouth disease virions into 12S particles as assessed by novel ELISAs specific for either 146S or 12S particles

Cited by 28 publications

References 33 publications

Identification of the Structural Basis of Thermal Lability of a Virus Provides a Rationale for Improved Vaccines

Identification of the Structural Basis of Thermal Lability of a Virus Provides a Rationale for Improved Vaccines

Isolation of Single-Domain Antibody Fragments That Preferentially Detect Intact (146S) Particles of Foot-and-Mouth Disease Virus for Use in Vaccine Quality Control

Structural and molecular basis for foot-and-mouth disease virus neutralization by two potent protective antibodies

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