Development of Cys38 knock-out and humanized version of NbAahII10 nanobody with improved neutralization of AahII Scorpion toxin

Rahma Ben Abderrazek, Cécile Vincke, Issam Hmila, Dirk Saerens, Naima Abidi, Mohamed El Ayeb, Serge Muyldermans, Balkiss Bouhaouala-Zahar

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

During scorpion envenoming, highly toxic small polypeptides of the venom diffuse rapidly within the victim, causing serious medical problems. Nanobodies (Nbs), the recombinant single-domain antigen-binding fragments of camel-specific heavy-chain only antibodies, offer special advantages in therapy over classic antibody fragments due to their robustness and smaller size, matching the size of the scorpion toxins. Recently, a potent AahII scorpion toxin-neutralizing Nb was identified. However, this NbAahII10 contains a single Cys in its first antigen-binding loop, leading to Nb dimerization upon prolonged storage. In this work, we first investigate the efficacy of NbAahII10 variants in which this Cys was substituted by Ala, Ser or Thr. Second, the NbAahII10 Cys/Ser mutant displaying the best functional properties is subsequently humanized. It is demonstrated that the maximally humanized version of NbAahII10 Cys/Ser maintains its high affinity for the antigen without conceding much on expression yield and stability. More importantly, its neutralizing capacity is preserved as all mice survive injections of seven LD50 and 50 of mice survived nine LD50 of the scorpion toxin. Thus, this humanized Nb is the best candidate to develop a therapy in human against the most toxic venom compound of one of the most dangerous scorpions.

Original languageEnglish
Pages (from-to)727-735
Number of pages9
JournalProtein Engineering, Design and Selection
Volume24
Issue number9
DOIs
Publication statusPublished - 1 Sep 2011
Externally publishedYes

Fingerprint

Single-Domain Antibodies
Scorpions
Antigens
Poisons
Venoms
Antibodies
Lethal Dose 50
Immunoglobulin Fragments
Dimerization
Polypeptides
Medical problems
Camelus
Peptides
Injections
Therapeutics

Keywords

  • anti-venom
  • humanization
  • nanobody
  • scorpion toxin
  • single-domain antibody

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Bioengineering
  • Molecular Biology

Cite this

Development of Cys38 knock-out and humanized version of NbAahII10 nanobody with improved neutralization of AahII Scorpion toxin. / Ben Abderrazek, Rahma; Vincke, Cécile; Hmila, Issam; Saerens, Dirk; Abidi, Naima; El Ayeb, Mohamed; Muyldermans, Serge; Bouhaouala-Zahar, Balkiss.

In: Protein Engineering, Design and Selection, Vol. 24, No. 9, 01.09.2011, p. 727-735.

Research output: Contribution to journalArticle

Ben Abderrazek, Rahma ; Vincke, Cécile ; Hmila, Issam ; Saerens, Dirk ; Abidi, Naima ; El Ayeb, Mohamed ; Muyldermans, Serge ; Bouhaouala-Zahar, Balkiss. / Development of Cys38 knock-out and humanized version of NbAahII10 nanobody with improved neutralization of AahII Scorpion toxin. In: Protein Engineering, Design and Selection. 2011 ; Vol. 24, No. 9. pp. 727-735.
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