Crowd on a Chip: Label-Free Human Monoclonal Antibody Arrays for Serotyping Influenza

Zhang, Hanyuan; Henry, Carole; Anderson, Christopher; Nogales, Aitor; DeDiego, Marta L.; Bucukovski, Joseph; Martínez-Sobrido, Luis; Wilson, Patrick C.; Topham, David J.; Miller, Benjamin L.

doi:10.1021/acs.analchem.8b02479

Cited by 19 publications

(29 citation statements)

References 51 publications

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“…Arrayed Imaging Reflectometry (AIR) has been used to provide information about anti-HA antibody titers in sera against different HA subtypes [38]. More recently, it has been used to demonstrate the multiplex capability of antibody arrays for live influenza virus characterization [39]. Here, we use AIR as a high-throughput screening approach to evaluate the cross-reactivity of a panel of recombinant hmAbs cloned from plasmablasts of volunteers vaccinated with different H3N2 strains (A/Perth/16/2009, A/Wisconsin/67/2005, A/Texas/50/ 2012, and A/Uruguay/716/2007) [39] against H3N2 and H3N8 CIVs and antigenically similar (HK68) or distinct (A/Wisconsin/67/2005, WI05) human H3N2 viruses.…”

Section: Protecting Naïve Populationsmentioning

confidence: 99%

Characterizing Emerging Canine H3 Influenza Viruses

et al. 2020

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Section: Protecting Naïve Populationsmentioning

confidence: 99%

Characterizing Emerging Canine H3 Influenza Viruses

et al. 2020

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“…Numbers on the left indicate the size of molecular markers for proteins (in kilodaltons). these technologies could also be used to assess the blocking capacity of NAbs for the treatment of viral infections or to test the efficacy of novel vaccine approaches (35,36,55,78), including universal influenza vaccine studies. In this study, we demonstrated the feasibility of using BIRFLU to evaluate the therapeutic value of antiviral drugs (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Neutralizing antibodies (NAbs) are the desired immunological outcome for induction of protective immunity after influenza vaccination (53)(54)(55)(56)(57)(58). However, the majority of assays to test for antibody-mediated protection efficacy in virus neutralization assays usually involve secondary methods to detect the presence of the virus.…”

Section: Birflu Reporter Gene Expression Levels Correlate With Dose-dmentioning

confidence: 99%

A Novel Fluorescent and Bioluminescent Bireporter Influenza A Virus To Evaluate Viral Infections

Nogales

Ávila‐Pérez

Rangel-Moreno

et al. 2019

J Virol

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Studying influenza A virus (IAV) requires the use of secondary approaches to detect the presence of virus in infected cells. To overcome this problem, we and others have generated recombinant IAV expressing fluorescent or luciferase reporter genes. These foreign reporter genes can be used as valid surrogates to track the presence of virus. However, the limited capacity for incorporating foreign sequences in the viral genome forced researchers to select a fluorescent or a luciferase reporter gene, depending on the type of study. To circumvent this limitation, we engineered a novel recombinant replication-competent bireporter IAV (BIRFLU) expressing both fluorescent and luciferase reporter genes. In cultured cells, BIRFLU displayed growth kinetics comparable to those of wild-type (WT) virus and was used to screen neutralizing antibodies or compounds with antiviral activity. The expression of two reporter genes allows monitoring of viral inhibition by fluorescence or bioluminescence, overcoming the limitations associated with the use of one reporter gene as a readout. In vivo, BIRFLU effectively infected mice, and both reporter genes were detected using in vivo imaging systems (IVIS). The ability to generate recombinant IAV harboring multiple foreign genes opens unique possibilities for studying virus-host interactions and for using IAV in high-throughput screenings (HTS) to identify novel antivirals that can be incorporated into the therapeutic armamentarium to control IAV infections. Moreover, the ability to genetically manipulate the viral genome to express two foreign genes offers the possibility of developing novel influenza vaccines and the feasibility for using recombinant IAV as vaccine vectors to treat other pathogen infections. IMPORTANCE Influenza A virus (IAV) causes a human respiratory disease that is associated with significant health and economic consequences. In recent years, the use of replication-competent IAV expressing an easily traceable fluorescent or luciferase reporter protein has significantly contributed to progress in influenza research. However, researchers have been forced to select a fluorescent or a luciferase reporter gene due to the restricted capacity of the influenza viral genome for including foreign sequences. To overcome this limitation, we generated, for the first time, a recombinant replication-competent bireporter IAV (BIRFLU) that stably expresses two reporter genes (one fluorescent and one luciferase) to track IAV infections in vitro and in vivo. The combination of cutting-edge techniques from molecular biology, animal research, and imaging technologies brings researchers the unique opportunity to use this new generation of reporter-expressing IAV to study viral infection dynamics in both cultured cells and animal models of viral infection. RESULTSGeneration of a BIRFLU expressing NLuc and Venus. To generate replicationcompetent bireporter IAV (BIRFLU) (Fig. 1), the sequence of NLuc and the porcine teschovirus 1 (PTV-1) 2A autoproteolytic cleavage site were cloned in front ...

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“…For example, both the H1N1 A/WSN/1933 and H3N2 A/Fujian/411/2002 strains show identical neutralization profiles against the six HA head antibodies and consequently have similar coordinates near (-0.6,1.2) in the HA head map ( Figure S2). While rare antibodies can inhibit both H1N1 and H3N2 viruses (18,20), if an antibody exists that neutralizes these two strains differently, it would split up the locations of these two viruses.…”

Section: Discussionmentioning

confidence: 99%

Using Neutralization Landscapes to enumerate Antibody Behavior and Decompose Antibody Mixtures

Einav

Creanga

Kanekiyo

2020

Preprint

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Antibodies constitute a key line of defense against invading pathogens. Although monoclonal antibodies can be readily quantified in terms of their binding affinity, neutralization potential, and antigenic site, there has been limited success in characterizing the individual antibodies within polyclonal sera. Dissecting the behavior of individual antibodies would improve our ability to compare sera — in both cross-sectional and longitudinal studies — and clarify how they differ. Using the neutralization data of monoclonal antibodies against the influenza virus, we develop a new form of antigenic cartography called a neutralization map that characterizes all possible antibody-virus interactions for the hemagglutinin head and stem. With these maps, the collective neutralization of polyclonal mixtures can be decomposed to determine the number of head and stem antibodies as well as their individual neutralization profiles. Using this decomposition, we find that over 70% of ferrets infected with a single strain of influenza elicit a polyclonal antibody response, and we demonstrate that serum titers against a subset of viruses can predict the titers of the remaining viruses. Looking forward, we anticipate that this framework can be used to both visually and quantitatively assess changes in the neutralization capacity of polyclonal serum elicited by natural infection or vaccination.

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Crowd on a Chip: Label-Free Human Monoclonal Antibody Arrays for Serotyping Influenza

Cited by 19 publications

References 51 publications

Characterizing Emerging Canine H3 Influenza Viruses

Characterizing Emerging Canine H3 Influenza Viruses

A Novel Fluorescent and Bioluminescent Bireporter Influenza A Virus To Evaluate Viral Infections

Using Neutralization Landscapes to enumerate Antibody Behavior and Decompose Antibody Mixtures

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