A recombinant single‐chain antibody fragment that neutralizes toxin II from the venom of the scorpion <i>Androctonus australis hector</i>

Mousli, Mohamed; Devaux, Christiane; Rochat, Hervé; Goyffon, M.; Billiald, Philippe

doi:10.1016/s0014-5793(98)01647-0

Cited by 58 publications

(51 citation statements)

References 34 publications

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“…Second, animals previously immunized with specific toxins were employed as source of antibodies. Several monoclonal antibodies have been generated by immunization of mice (23,32,33). By means of recombinant DNA technology, different simplified antibody formats (scFv, Fab, dimer, and tandem repeats) have been constructed (34 -38).…”

Section: Discussionmentioning

confidence: 99%

Exploiting Cross-reactivity to Neutralize Two Different Scorpion Venoms with One Single Chain Antibody Fragment

Riaño-Umbarila

Contreras-Ferrat

Olamendi-Portugal

et al. 2011

Journal of Biological Chemistry

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We report the optimization of a family of human single chain antibody fragments (scFv) for neutralizing two scorpion venoms. The parental scFv 3F recognizes the main toxins of Centruroides noxius Hoffmann (Cn2) and Centruroides suffusus suffusus (Css2), albeit with low affinity. This scFv was subjected to independent processes of directed evolution to improve its recognition toward Cn2 (Riaño-Umbarila, L., Juárez-González, V. R., Olamendi-Portugal, T., Ortíz-León, M., Possani, L. D., and Becerril, B. (2005) FEBS J. 272, 2591-2601) and Css2 (this work). Each evolved variant showed strong cross-reactivity against several toxins, and was capable of neutralizing Cn2 and Css2. Furthermore, each variant neutralized the whole venoms of the above species. As far as we know, this is the first report of antibodies with such characteristics. Maturation processes revealed key residue changes to attain expression, stability, and affinity improvements as compared with the parental scFv. Combination of these changes resulted in the scFv LR, which is capable of rescuing mice from severe envenomation by 3 LD 50 of freshly prepared whole venom of C. noxius (7.5 g/20 g of mouse) and C. suffusus (26.25 g/20 g of mouse), with surviving rates between 90 and 100%. Our research is leading to the formulation of an antivenom consisting of a discrete number of human scFvs endowed with strong cross-reactivity and low immunogenicity.

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Section: Discussionmentioning

confidence: 99%

Exploiting Cross-reactivity to Neutralize Two Different Scorpion Venoms with One Single Chain Antibody Fragment

Riaño-Umbarila

Contreras-Ferrat

Olamendi-Portugal

et al. 2011

Journal of Biological Chemistry

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“…The purity of product was analyzed by vertical sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) according to Laemmli [22] using a 10 % polyacrylamide gel under non-reducing conditions. Then, all samples were used at the concentration of 500 µg/ml.…”

Section: Purity Evaluationmentioning

confidence: 99%

Optimization of purification procedure for horse F(ab´)2 antivenom against <i>Androctonus crassicauda</i> (Scorpion) venom

Taherian¹,

Fazilati²,

Moghadam³

et al. 2018

Trop. J. Pharm Res

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“…7. The periplas-* This work was supported in part by grants from the Centre National de la Recherche Scientifique.…”

Section: Bacterial Expression Of Scfv-6h8mentioning

confidence: 99%

scFv Single Chain Antibody Variable Fragment as Inverse Agonist of the β2-Adrenergic Receptor

Peter

Eftekhari²,

Billiald

et al. 2003

Journal of Biological Chemistry

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Antibodies directed against the second extracellular loop of G protein-coupled receptors were shown to possess functional activities. Using a functional monoclonal antibody against the human ␤ 2 -adrenergic receptor, a scFv fragment with high affinity for the target epitope was constructed and produced. The fragment recognized the ␤ 2 -adrenergic receptors on A431 cells, blocked cAMP accumulation induced by the ␤ 2 -agonist salbutamol, and decreased basal cAMP accumulation in the same cells. Their in vitro activity was tested on neonatal rat cardiomyocytes. The antibody fragments blocked the chronotropic activity induced by the ␤ 2 -agonist clenbuterol. They also decreased the in vivo heart beating frequency of mice pretreated with bisoprolol (a ␤ 1 -adrenergic receptor antagonist) for 4 min after injection. The immunological approach presented here may serve as a strategy for the synthesis of a new class of allosteric modulators for G protein-coupled receptors.The G protein-coupled receptor family is one of the main targets of currently used drugs (1). Most of the structural insights into this family of receptors were obtained from studies of the ␤ 2 -adrenergic receptor, the first of this family of neurotransmitter receptors to be cloned and sequenced. This receptor is an integral membrane protein consisting of seven membrane spanning ␣-helices, which form a pharmacophore pocket, linked together by extra-and intracellular loops (2). One of the pharmacological challenges posed by this family of receptors is the presence of multiple subtypes, all recognizing the same endogenous ligands. This suggests a high conservation of the pharmacophore for a particular family of receptors, thus explaining the difficulty to synthesize drugs (agonists or antagonists), specific for one of these subtypes. Autoantibodies directed against cardiovascular G protein coupled receptors, functionally interfering with the target, have been described in several cardiovascular diseases. Most of these autoantibodies are directed against the second extracellular loop and are exquisitely specific for one of the receptor subtypes in view of the highly variable structure of this domain (3). Lebesgue et al. (4) reported the selection of monoclonal antibodies directed against a synthetic peptide whose sequence was derived from the second extracellular loop of G protein-coupled receptors. The selected monoclonal antibody against the ␤ 2 -adrenergic receptor had partial agonist activity as a dimer and antagonist activity as a monovalent Fab fragment (4, 5). This antibody was used to construct a scFv fragment (single chain variable fragment), which was cloned, sequenced, and expressed in Escherichia coli. In this study, we describe the sequence, the immunochemical, pharmacological, and physiological properties of this scFv fragment. These results open the way for the development of new strategies to synthesize molecules, which are highly specific for one of the subtypes of a particular G proteincoupled receptor family and can allosterically modul...

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A recombinant single‐chain antibody fragment that neutralizes toxin II from the venom of the scorpion Androctonus australis hector

Cited by 58 publications

References 34 publications

Exploiting Cross-reactivity to Neutralize Two Different Scorpion Venoms with One Single Chain Antibody Fragment

Exploiting Cross-reactivity to Neutralize Two Different Scorpion Venoms with One Single Chain Antibody Fragment

Optimization of purification procedure for horse F(ab´)2 antivenom against <i>Androctonus crassicauda</i> (Scorpion) venom

scFv Single Chain Antibody Variable Fragment as Inverse Agonist of the β2-Adrenergic Receptor

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