Standardized guidelines for Africanized honeybee venom production needed for development of new apilic antivenom

ABSTRACT

Africanized bees have spread across the Americas since 1956 and consequently resulted in human and animal deaths attributed to massive attacks related to exposure from Argentina to the USA. In Brazil, more than 100,000 accidents were registered in the last 5 years with a total of 303 deaths. To treat such massive attacks, Brazilian researchers developed the first specific antivenom against Africanized honey bee sting exposure. This unique product, the first of its kind in the world, has been safely tested in 20 patients during a Phase 2 clinical trial. To develop the antivenom, a standardized process was undertaken to extract primary venom antigens from the Africanized bees for immunization of serum-producing horses. This process involved extracting, purifying, fractionating, characterizing, and identifying the venom (apitoxin) employing mass spectrometry to generate standardized antigen for hyperimmunization of horses using the major toxins (melittin and its isoforms and phospholipase A2). The current guide describes standardization of the entire production chain of venom antigens in compliance with good manufacturing practices (GMP) required by regulatory agencies. Emphasis is placed upon the welfare of bees and horses during this process, as well as the development of a new biopharmaceutical to ultimately save lives.

KEYWORDS:

• Africanized honeybee
• Apis mellifera
• good manufacturing practices
• hyperimmunization
• standard operating procedures

Acknowledgments

The authors convey special thanks to the Center for the Study of Venoms and Venomous Animals (CEVAP/UNESP), College of Veterinary Medicine and Animal Sciences (FMVZ-UNESP), and Butantan and Vital Brazil Institutes for providing the resources that contributed to this work.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Authorship contributions

All authors are responsible for the writing, content, and conclusions in this article.

Ethics approval

All animal procedures and regular infrastructure of the Center for the Study of Venoms and Venomous Animals (CEVAP/UNESP) were approved by the Animal Use Ethics Committee of Biotechnology Institute (IBTEC), São Paulo State University (UNESP), Botucatu, São Paulo, Brazil, Proc. No. 18/2018.

Additional information

Funding

This study was supported by the National Council of Technological and Scientific Development (CNPq) under Grants No. 563582/2010-3 (BB), and 441463/2019-3 (BB), Coordination for the Improvement of Higher Education Personnel (CAPES - AUX-PE Toxinology) under Grants No. 23038.006285/2011-21 and No. 23038.008557/2010 (BB), and The São Paulo Research Foundation (FAPESP) under Grants No. 2021/11936-3 (BB). RSF Jr. and ROO are a CNPq PQ-1D fellow researchers respectively No. 301608/2022-9, and No. 304720/2020-8; BB and DCP are a CNPq PQ-2 fellow researchers respectively No. 306339/2020-0 and No. 301974/2019-5.