Next Article in Journal
Antipruritic Effects of Botulinum Neurotoxins
Next Article in Special Issue
Antivenom Production against Bothrops jararaca and Bothrops erythromelas Snake Venoms Using Cross-Linked Chitosan Nanoparticles as an Immunoadjuvant
Previous Article in Journal
Dose-Response Modelling of Paralytic Shellfish Poisoning (PSP) in Humans
Article Menu
Issue 4 (April) cover image

Export Article

Open AccessFeature PaperArticle
Toxins 2018, 10(4), 142;

Camelid Single-Domain Antibodies (VHHs) against Crotoxin: A Basis for Developing Modular Building Blocks for the Enhancement of Treatment or Diagnosis of Crotalic Envenoming

Fundação Oswaldo Cruz, Fiocruz Rondônia, Porto Velho, 76812-245 Rondônia, Brazil
Instituto Federal de Educação, Ciência e Tecnologia de Rondônia, IFRO, Guajará-Mirim, 76850-000 Rondônia, Brazil
Universidade Federal Fluminense, UFF, Rio de Janeiro, 24220-900 Rio de Janeiro, Brazil
Embrapa Agroindústria Tropical, Fortaleza, 60020-181 Ceará, Brazil
Universidade Federal de Rondônia, UNIR, Porto Velho, 76801-974 Rondônia, Brazil
Centro Universitário São Lucas, UniSL, 76812-245 Porto Velho-RO, Brazil
Plataforma Bi-institucional de Pesquisa em Medicina Translacional, Fiocruz-USP, Ribeirão Preto, 14049-900 São Paulo, Brazil
Departamento de Medicina da Universidade Federal de São Carlos, Demed-UFSCAR, São Carlos, 13565-905 São Paulo, Brazil
Authors to whom correspondence should be addressed.
Received: 1 February 2018 / Revised: 11 March 2018 / Accepted: 16 March 2018 / Published: 29 March 2018
(This article belongs to the Special Issue Discovery of Antibodies and Novel Antivenoms against Envenoming)
Full-Text   |   PDF [39312 KB, uploaded 3 May 2018]   |  


Toxic effects triggered by crotalic envenoming are mainly related to crotoxin (CTX), composed of a phospholipase A2 (CB) and a subunit with no toxic activity (CA). Camelids produce immunoglobulins G devoid of light chains, in which the antigen recognition domain is called VHH. Given their unique characteristics, VHHs were selected using Phage Display against CTX from Crotalus durissus terrificus. After three rounds of biopanning, four sequence profiles for CB (KF498602, KF498603, KF498604, and KF498605) and one for CA (KF498606) were revealed. All clones presented the VHH hallmark in FR2 and a long CDR3, with the exception of KF498606. After expressing pET22b-VHHs in E. coli, approximately 2 to 6 mg of protein per liter of culture were obtained. When tested for cross-reactivity, VHHs presented specificity for the Crotalus genus and were capable of recognizing CB through Western blot. KF498602 and KF498604 showed thermostability, and displayed affinity constants for CTX in the micro or nanomolar range. They inhibited in vitro CTX PLA2 activity, and CB cytotoxicity. Furthermore, KF498604 inhibited the CTX-induced myotoxicity in mice by 78.8%. Molecular docking revealed that KF498604 interacts with the CA–CB interface of CTX, seeming to block substrate access. Selected VHHs may be alternatives for the crotalic envenoming treatment. View Full-Text
Keywords: crotoxin; CB; VHH; Crotalus durissus terrificus crotoxin; CB; VHH; Crotalus durissus terrificus

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

Share & Cite This Article

MDPI and ACS Style

Luiz, M.B.; Pereira, S.S.; Prado, N.D.R.; Gonçalves, N.R.; Kayano, A.M.; Moreira-Dill, L.S.; Sobrinho, J.C.; Zanchi, F.B.; Fuly, A.L.; Fernandes, C.F.; Zuliani, J.P.; Soares, A.M.; Stabeli, R.G.; Fernandes, C.F.C. Camelid Single-Domain Antibodies (VHHs) against Crotoxin: A Basis for Developing Modular Building Blocks for the Enhancement of Treatment or Diagnosis of Crotalic Envenoming. Toxins 2018, 10, 142.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics



[Return to top]
Toxins EISSN 2072-6651 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top