Grouping of monoclonal antibodies to adenovirus hexons by their cross-reactivity

Ádám, Éva; Lengyel, A; Takács, Mária; Erdei, J.; Fachet, J; Nász, I

doi:10.1007/bf01310543

Cited by 11 publications

(6 citation statements)

References 14 publications

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“…In the past, either purified virus or purified virus proteins were used for immunization to generate hexon-specific antibodies [ 44 , 66 ]. Since in our hands, purified MAdVs yields were lower compared to HAdVs and purification of MAdV hexon protein from lysates of infected cells was not efficient (not shown), we decided to produce recombinant hexon fragments containing HVRs to induce specific antibodies.…”

Section: Resultsmentioning

confidence: 99%

“…Standard procedures to generate adenovirus hexon-specific antibodies include preparations of purified virions, or viral proteins for the purpose of immunization [ 44 , 66 ]. However, not all adenoviruses can be amplified efficiently in cell culture.…”

Section: Discussionmentioning

confidence: 99%

“…Hexon antigenic determinants and antibodies raised against hexon will not only be influenced by the nature of antigen used for immunization, e.g., purified virus, or hexon or partial fragment, but also by the immunization procedure which can include use of alum precipitation, adjuvants, and terminal boost with virus only. Thus, generation of mouse monoclonal antibodies resulted in cross-reactive genus-, species- or type-specific antibodies [ 44 , 66 ]. Type-specific and neutralizing epitopes have been suggested to be conformation dependent [ 27 , 34 ], whereas non-neutralizing and cross-reactive hexon antibodies with common species- and genus-specific reactivity have been suggested to recognize sites that are accessible on the purified hexon but partially masked in the intact virion and outside of the HVRs [ 27 , 44 , 45 ].…”

Section: Discussionmentioning

confidence: 99%

“…In contrast to these findings, other groups reported generation of neutralizing antibodies using small HVR peptides only [ 28 , 43 ]. Non-neutralizing and cross-reactive hexon antibodies with common species- and genus-specific reactivity have been suggested to recognize sites that are accessible on the purified hexon but partially masked in the intact virion and outside of the HVRs [ 27 , 44 ]. Such antibodies were characterized using complement fixation, immunodiffusion, immunoprecipitation and immunoblot assays [ 45 – 47 ].…”

Section: Introductionmentioning

confidence: 99%

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Small-size recombinant adenoviral hexon protein fragments for the production of virus-type specific antibodies

Pačesa

Hendrickx

Bieri

et al. 2017

Virol J

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BackgroundAdenoviruses are common pathogens infecting animals and humans. They are classified based on serology, or genome sequence information. These methods have limitations due to lengthy procedures or lack of infectivity data. Adenoviruses are easy to produce and amenable to genetic and biochemical modifications, which makes them a powerful tool for biological studies, and clinical gene-delivery and vaccine applications. Antibodies directed against adenoviral proteins are important diagnostic tools for virus identification in vivo and in vitro, and are used to elucidate infection mechanisms, often in combination with genomic sequencing and type specific information from hyper-variable regions of structural proteins.ResultsHere we describe a novel and readily useable method for cloning, expressing and purifying small fragments of hyper-variable regions 1-6 of the adenoviral hexon protein. We used these polypeptides as antigens for generating polyclonal rabbit antibodies against human adenovirus 3 (HAdV-B3), mouse adenovirus 1 (MAdV-1) and MAdV-2 hexon. In Western immunoblots with lysates from cells infected from thirteen human and three mouse viruses, these antibodies bound to homologous full-length hexon protein and revealed variable levels of cross-reactivity to heterologous hexons. Results from immuno-fluorescence and electron microscopy studies indicated that HAdV-B3 and MAdV-2 hexon antibodies recognized native forms of hexon.ConclusionsThe procedure described here can in principle be applied to any adenovirus for which genome sequence information is available. It provides a basis for generating novel type-specific tools in diagnostics and research, and extends beyond the commonly used anti-viral antibodies raised against purified viruses or subviral components.Electronic supplementary materialThe online version of this article (doi:10.1186/s12985-017-0822-5) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Small-size recombinant adenoviral hexon protein fragments for the production of virus-type specific antibodies

Pačesa

Hendrickx

Bieri

et al. 2017

Virol J

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“…The monoclonal antibody, 1A3, was prepared against purified human Ad serotype 1 hexon molecule as described previously [20].…”

Section: Production Of the Hexon-specific Monoclonal Antibody 1a3mentioning

confidence: 99%

Screening adenoviruses in stool samples: evaluation of a genus-specific monoclonal antibody based enzyme immunoassay

Bányai

Máté

Ádám

et al. 2003

Acta Microbiologica et Immunologica Hungarica

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To screen fecal samples for adenovirus antigens a genus-specific monoclonal antibody based enzyme immunoassay was developed. In a comparative analysis with commercial latex agglutination test, high sensitivity was demonstrated. The assay did not detect other viruses usually found in faeces suggesting its specificity. One hundred and eighty stool samples collected in Baranya County were tested and 13 (7.2%) of them showed reactivity. The application of our immunoassay combined with other, more sophisticated methods may help us to determine the serotype specificity of these adenovirus isolates and assess the importance of adenoviruses in viral gastroenteritis.

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Preparation and characterization of monoclonal antibodies to enteric adenovirus types 40 and 41

Herrmann

Perron-Henry

Stobbs-Walro

et al. 1987

Archives of Virology

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We have prepared monoclonal antibodies to each of the enteric adenoviruses types 40 and 41. Three different hybridoma cell lines were selected which produced antibody found to react by radioimmunoprecipitation with adenovirus (Ad) hexon antigens. One was specific for Ad 40, another for Ad 41, and a third one reacted with both types. When tested in an enzyme immunoassay against all 41 known human Ad types, the type-specific monoclonal antibody against Ad 40 reacted homotypically, as did the monoclonal antibody against Ad 41. In addition, these monoclonal antibodies neutralized the homologous enteric Ad type. The monoclonal antibody which reacted with both enteric Ad types also showed lower levels of reactivity with the group C adenoviruses types 2, 5, and 6. The monoclonal antibodies produced will provide a definitive means for rapid identification of specific Ad types, and will be useful in defining the relationship of enteric adenoviruses to other types.

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Grouping of monoclonal antibodies to adenovirus hexons by their cross-reactivity

Cited by 11 publications

References 14 publications

Small-size recombinant adenoviral hexon protein fragments for the production of virus-type specific antibodies

Small-size recombinant adenoviral hexon protein fragments for the production of virus-type specific antibodies

Screening adenoviruses in stool samples: evaluation of a genus-specific monoclonal antibody based enzyme immunoassay

Preparation and characterization of monoclonal antibodies to enteric adenovirus types 40 and 41

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