Passive immunization of guinea pigs with llama single-domain antibody fragments against foot-and-mouth disease

Harmsen, Michiel M.; Solt, C. B. van; Fijten, H.P.D.; Keulen, Lucien van; Rosalia, Rodney Alexander; Weerdmeester, K.; Cornelissen, A.H.M.; Bruin, M.G.M. de; Eblé, P.L.; Dekker, A.

doi:10.1016/j.vetmic.2006.10.029

Cited by 64 publications

(78 citation statements)

References 29 publications

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“…23,62 Unlike conventional VHs, the hydrophilic mutations in the VHHs contribute to high-expression yields. 26,45 The high refolding capability of VHHs has also been correlated with a high production level in E. coli (up to 10 mg/L). 23,40,61 Despite the satisfactory yields, expression levels of different clones can vary by a factor of 100; 26,45 the use of random molecular evolution using DNA shuffling may improve production.…”

Section: Nb Expression Systemsmentioning

confidence: 99%

“…26,45 The high refolding capability of VHHs has also been correlated with a high production level in E. coli (up to 10 mg/L). 23,40,61 Despite the satisfactory yields, expression levels of different clones can vary by a factor of 100; 26,45 the use of random molecular evolution using DNA shuffling may improve production. 21 Attempts to use filamentous fungi resulted in limited proteolytic degradation of the secreted product, 36 possibly due to the …”

Section: Nb Expression Systemsmentioning

confidence: 99%

“…45 Conjugation to epidermal growth factor receptor for solid tumor inhibition. 46 Specific enzyme inhibitory activity in vivo.…”

mentioning

confidence: 99%

“…68 High yields can be obtained from yeast systems, like Saccharomyces cerevisiae. 38,69 These systems may also coproduce N-glycosylated Nbs with yeastspecific high-mannose oligosaccharides 26,45 which, due to their binding to specific mannose receptors on reticuloendothelial cells, 70 could increase the in vivo immunogenicity and decrease the serum half-life of the moiety.…”

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confidence: 99%

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Nanobodies as novel agents for disease diagnosis and therapy

Siontorou¹

2013

IJN

123

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Section: Nb Expression Systemsmentioning

confidence: 99%

Section: Nb Expression Systemsmentioning

confidence: 99%

“…45 Conjugation to epidermal growth factor receptor for solid tumor inhibition. 46 Specific enzyme inhibitory activity in vivo.…”

mentioning

confidence: 99%

mentioning

confidence: 99%

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Nanobodies as novel agents for disease diagnosis and therapy

Siontorou¹

2013

IJN

123

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“…For instance, VHHs provided full protection of mice challenged with lethal doses of a small (»7 kDa) scorpion toxin, 13 but potent in vitro neutralizing VHHs failed to provide protection when tested in vivo in guinea pigs challenged with food-and-mouth disease virus. 14 Different strategies have been devised to improve the performance of neutralizing VHHs. Commonly, one or more VHHs are combined in tandem to produce bivalent or bispecific constructs.…”

Section: Introductionmentioning

confidence: 99%

Increasing the potency of neutralizing single-domain antibodies by functionalization with a CD11b/CD18 binding domain

Rossotti

González-Techera

Guarnaschelli

et al. 2015

mAbs

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Recombinant single domain antibodies (nanobodies) constitute an attractive alternative for the production of neutralizing therapeutic agents. Their small size warrants rapid bioavailability and fast penetration to sites of toxin uptake, but also rapid renal clearance, which negatively affects their performance. In this work, we present a new strategy to drastically improve the neutralizing potency of single domain antibodies based on their fusion to a second nanobody specific for the complement receptor CD11b/CD18 (Mac-1). These bispecific antibodies retain a small size (»30 kDa), but acquire effector functions that promote the elimination of the toxin-immunocomplexes. The principle was demonstrated in a mouse model of lethal toxicity with tetanus toxin. Three anti-tetanus toxin nanobodies were selected and characterized in terms of overlapping epitopes and inhibition of toxin binding to neuron gangliosides. Bispecific constructs of the most promising monodomain antibodies were built using anti Mac-1, CD45 and MHC II nanobodies. When co-administered with the toxin, all bispecific antibodies showed higher toxin-neutralizing capacity than the monomeric ones, but only their fusion to the anti-endocytic receptor Mac-1 nanobody allowed the mice to survive a 10-fold lethal dose. In a model of delayed neutralization of the toxin, the anti-Mac-1 bispecific antibodies outperformed a sheep anti-toxin polyclonal IgG that had shown similar neutralization potency in the co-administration experiments. This strategy should have widespread application in the development of nanobody-based neutralizing therapeutics, which can be produced economically and more safely than conventional antisera.

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Single‐Domain Antibodies and Their Utility

2013

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Vous avez des questions? Nous pouvons vous aider. Pour communiquer directement avec un auteur, consultez la première page de la revue dans laquelle son article a été publié afin de trouver ses coordonnées. Si vous n'arrivez pas à les repérer, communiquez avec nous à PublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. NRC Publications Record / Notice d'Archives des publications de CNRC:http://nparc.cisti-icist.nrc-cnrc.gc.ca/eng/view/object/?id=e552ed0a-502f-47b3-a6f6-9daf5bd955d3 http://nparc.cisti-icist.nrc-cnrc.gc.ca/fra/voir/objet/?id=e552ed0a-502f-47b3-a6f6-9daf5bd955d3 ABSTRACTEngineered monoclonal antibody fragments have gained market attention due to their versatility and tailor-made potential and are now considered to be an important part of future immunobiotherapeutics. Single-domain antibodies (sdAbs), also known as nanobodies, are derived from VHHs [variable domains (V) of heavy-chain-only antibodies (HCAb)] of camelid heavy-chain antibodies. These nature-made sdAbs are well suited for various applications due to their favorable characteristics such as small size, ease of genetic manipulation, high affinity and solubility, overall stability, resistance to harsh conditions (e.g., low pH, high temperature), and low immunogenicity. Most importantly, sdAbs have the feature of penetrating into cavities and recognizing hidden epitopes normally inaccessible to conventional antibodies, mainly due to their protruding CDR3/H3 loops. In this unit, we will present and discuss comprehensive and step-by-step protocols routinely practiced in our laboratory for isolating sdAbs from immunized llamas (or other members of the Camelidae family) against target antigens using phage-display technology. Expression, purification, and characterization of the isolated sdAbs will then be described, followed by presentation of several examples of applications of sdAbs previously characterized in our laboratory and elsewhere.

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Passive immunization of guinea pigs with llama single-domain antibody fragments against foot-and-mouth disease

Cited by 64 publications

References 29 publications

Nanobodies as novel agents for disease diagnosis and therapy

Nanobodies as novel agents for disease diagnosis and therapy

Increasing the potency of neutralizing single-domain antibodies by functionalization with a CD11b/CD18 binding domain

Single‐Domain Antibodies and Their Utility

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