Hoffmeister Series Ions Protect Diphtheria Toxoid from Structural Damages at Solvent/Water Interface

Materials

Jocimara A.M. Namur³  Célia S Takata¹  Pedro S. de Araujo² 

[1] Divisão de Desenvolvimento Tecnológico e Produção-I. Butantan Av. Vital Brasil, 1500 (05503-900) Butantan, São Paulo, SP, Brasil; E-Mail:;I. de Química, (05508-000) Universidade de São Paulo Av. Lineu Prestes 748, São Paulo, SP, Brasil; E-Mails:;Laboratório de Microesferas e Lipossomas – Centro de Biotecnologia- I. Butantan, Av. Vital Brasil, 1500, 05503-900 - Butantan, São Paulo, SP, Brasil

关键词: Hoffmeister series ions;    protein solubilization;    protein stabilization;    interaction protein/organic solvent;    protein microencapsulation;    adjuvant particulate;    adjuvant;   

DOI  :  10.3390/ma2030765

来源: mdpi

【 摘 要 】

During the W₁/O phase (in the W₁/O/W₂ process) of protein microencapsulation within poly-lactide-co-glycolide (PLGA), hydrophobic interfaces are expanded where interfacial adsorption occurs followed by protein unfolding and aggregation. Spectroscopic and immunological techniques were used to ascertain the effects of the Hoffmeister series ions on Diphtheria toxoid (Dtxd) stability during the W₁/O phase. A correlation was established between salts used in aqueous solutions and the changes in Dtxd solubility and conformation. The Dtxd α-helical content was quite stable thus leading to the conclusion that encapsulation was followed by protein aggregation, with minor exposition of hydrophobic residues and a small change at the S-S dihedral angle. Dtxd aggregation is 95% avoided by the chaotropic SCN^-. This was used to prepare a stable Dtxd and immunologically recognized/PLGA formulation in the presence of 30 mM SNC^-. The recovery increased by 10.42% or 23.2% when microencapsulation was within the -COOMe or -COOH (12kDa) PLGA, respectively. In conclusion, the aim of this work was achieved, which was to obtain the maximum of Dtxd stability after contact with CH₂Cl₂ to begin its PLGA microencapsulation within ideal conditions. This was a technological breakthrough because a simple solution like salt addition avoided heterologous proteins usage.

【 授权许可】

CC BY   
© 2009 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland.