Immunobiological role of llama heavy-chain antibodies against a bacterial β-lactamase

Ferrari, Alejandro; Rodríguez, María Margarita; Power, Pablo; Weill, Federico S.; Simone, Emilio A. De; Gutkind, Gabriel; Leoni, Juliana

doi:10.1016/j.vetimm.2007.03.003

Cited by 12 publications

(13 citation statements)

References 25 publications

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“…This issue has been partially addressed by Lauwereys et al (1998), who succeeded in demonstrating enzyme modulation by rVHH fragments for two enzymes (lysozyme and a-amylase); however, it is not clear whether these results may be extrapolated to all the other enzymes, or are a consequence of a special feature of the selected antigens. The relevance of this matter is reinforced by our own preliminary reports concerning b-lactamase activation by pHCAbs from a single serum sample of a llama (Lama glama; Ferrari et al, 2007).…”

Section: Introductionmentioning

confidence: 88%

“…Cloning, expression and purification were performed as previously described (Power et al, 2005;Ferrari et al, 2007).…”

Section: Methodsmentioning

confidence: 99%

“…Animals immunized with APM were inoculated once a week, and serum samples were collected at days 0 (preimmune), 8, 29 and 42. Animals immunized with CTX-M-2 were inoculated every 2 weeks and serum samples were collected at days 0 (preimmune), 60, 75, 90, 150, 170 and 215. Serum titration by ELISA The titration of total serum Igs was performed as previously described (Ferrari et al, 2007). Briefly, 96-well microplates (Nunc Maxisorp, Roskilde, Denmark) were coated with 0.1 mg of purified APM or CTX-M-2 per well, and blocked with 3% non-fat milk diluted in PBS.…”

Section: Methodsmentioning

confidence: 99%

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Activity modulation of microbial enzymes by llama (Lama glama) heavy-chain polyclonal antibodies during in vivo immune responses

Ferrari

Weill

Paz

et al. 2012

Animal

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Since they were first described in 1993, it was found that recombinant variable fragments (rVHHs) of heavy-chain antibodies (HCAbs) from Camelidae have unusual biophysical properties, as well as a special ability to interact with epitopes that are cryptic for conventional Abs. It has been assumed that in vivo raised polyclonal HCAbs (pHCAbs) should behave in a similar manner than rVHHs; however, this assumption has not been tested sufficiently. Furthermore, our own preliminary work on a single serum sample from a llama immunized with a b-lactamase, has suggested that pHCAbs have no special ability to down-modulate catalytic activity. In this work, we further explored the interaction of pHCAbs from four llamas raised against two microbial enzymes and analyzed it within a short and a long immunization plan. The relative contribution of pHCAbs to serum titer was found to be low compared with that of the most abundant conventional subisotype (IgG 1 ), during the whole immunization schedule. Furthermore, pHCAbs not only failed to inhibit the enzymes, but also activated one of them. Altogether, these results suggest that raising high titer inhibitory HCAbs is not a straightforward strategy -neither as a biotechnological strategy nor in the biological context of an immune response against infection -as raising inhibitory rVHHs.

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Section: Introductionmentioning

confidence: 88%

“…Cloning, expression and purification were performed as previously described (Power et al, 2005;Ferrari et al, 2007).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Activity modulation of microbial enzymes by llama (Lama glama) heavy-chain polyclonal antibodies during in vivo immune responses

Ferrari

Weill

Paz

et al. 2012

Animal

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“…Muchos autores han descripto que los dominios VHH de los HCAbs tienen importantes ventajas en comparación con las Igs convencionales, sin embargo las propiedades fisiológicas de la molécula de HCAbs en su totalidad no parece tener alguna ventaja cuando se compara con las Igs convencionales, (Ferrari et al, 2007). Además nosotros no fuimos capaces de demostrar funciones efectoras como fijación de complemento por parte de los HCAbs, cuando las Igs convencionales tienen sitios de fijación para C1q, (Saccodossi et al, 2012).…”

Section: Conclusiónunclassified

Caracterización de IgM, IgG Total, IgG1 y anticuerpos de cadena pesada en calostro de llamas ("Lama glama") mediante Elisa

Caggiano¹,

Saccodossi

Gentile

et al. 2014

Revista Complutense De Ciencias Veterinarias

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RESUMENObjetivos: determinar los niveles de IgM, IgG total y Anticuerpos de Cadena Pesada (HCAbs; por su sigla en inglés Heavy Chain Antibodies) (IgG2 e IgG3) en calostro de llamas y evaluar la concentración de HCAbs en relación a la IgG total y al isotipo convencional IgG1.Métodos: en este estudio se utilizaron 15 llamas preñadas, que fueron ordeñadas dentro de las primeras 24 horas post-parto. Se diseñaron ELISAs Sandwich para la cuantificación de IgM total, IgG total e IgG1. La concentración de HCAbs fue calculada mediante la diferencia entre IgG total e IgG1. (HCAbs =IgG total-IgG1).Resultados: los niveles encontrados fueron: IgM=17.02 mg/ml (DS=9.85) IgG total= 42.54 mg/ml (DS=27.79), IgG1=24.34 mg/ml (DS=13.96) y HCAbs (IgG total-IgG1)= 18.19 mg/ml (DS=15.49). Los resultados de IgG fueron consistentes en relación a lo descripto en otras especies con similar tipo de placentación. Los HCAbs representan el 43% de IgG calostral y la concentración de IgM presentó valores muy altos (dos y tres veces más elevados) que los descriptos en las especies con similar tipo de placentación.

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“…Other structural features of heavy-chain antibodies, which differentiate them from conventional antibodies, include a higher proportion of hydrophilic amino acids in variable domains (VHH/VNAR), the presence of disulfide bonds stabilizing the antigen-binding site, and an unusually high number of mutational hot spots responsible for structural variability of heavychain antibodies (Harmsen and de Haard 2007). In addition to structural studies, recent research has been focused also on the evolution of heavychain antibodies (Nguyen et al 2002;Flajnik et al 2011), their possible function in the immune system (Ferrari et al 2007;Saccodossi et al 2012), and molecular mechanisms of germline gene segment rearrangement (Nguyen et al 2000). Some investigators have also used heavy-chain antibodies as a suitable alternative to conventional polyclonal antibodies, mainly for diagnostic purposes (Anderson and Goldman 2008;Torigoe et al 2012).…”

Section: Heavy-chain Antibodies Of Camelids and Sharksmentioning

confidence: 99%

Single-domain antibody fragments derived from heavy-chain antibodies: a review

Eyer¹,

Hruška²

2012

Vet. Med.

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Single-domain antibody (sdAb) fragments derived from heavy-chain antibodies of camelids and cartilaginous fish represent a new generation of therapeutic agents and immunoreagents. Due to their unique characteristics, such as low molecular weight, high physical-chemical stability, good water solubility, and the ability to bind antigens inaccessible to conventional antibodies, they could potentially act as a substitute for conventional therapeutic drugs in the treatment of serious human diseases, and, moreover, could be broadly used in analyses and diagnostics. In this review article, an analysis of 826 publications oriented to heavy-chain antibodies and their sdAb fragments indexed in the Web of Science ® database since 1993 has been carried out. Attention has predominantly been paid to papers published from 2010 to June 2012. Key publications are presented in tables and are characterised by descriptive words, abstracts and references. The presented publications have been sorted according to seven basic criteria: review articles and monographs, heavy-chain antibodies of camelids and sharks, production of sdAb fragments using recombinant technology, characteristic properties of sdAb fragments, application of sdAb fragments in therapy, application of sdAb fragments in diagnostic and immunoanalytical methods and other prospective uses of sdAb fragments. This review article should highlight the typical properties of heavy-chain antibodies and sdAb fragments which differentiate them from conventional antibodies and other available recombinant fragments, and also emphasize their extremely broad application potential, mainly in human disease therapy. At the same time it allows an easy and rapid orientation in numerous publications written on this subject, and facilitates the search for the required data.

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Immunobiological role of llama heavy-chain antibodies against a bacterial β-lactamase

Cited by 12 publications

References 25 publications

Activity modulation of microbial enzymes by llama (Lama glama) heavy-chain polyclonal antibodies during in vivo immune responses

Activity modulation of microbial enzymes by llama (Lama glama) heavy-chain polyclonal antibodies during in vivo immune responses

Caracterización de IgM, IgG Total, IgG1 y anticuerpos de cadena pesada en calostro de llamas ("Lama glama") mediante Elisa

Single-domain antibody fragments derived from heavy-chain antibodies: a review

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