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

Autor(es)

Namur, Jocimara A. M. ^{[1, 2]} ; Takata, Celia S. ^[3] ; de Araujo, Pedro S. ^[1] ; Bueno-da-Costa, Maria H. ^[2]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, BR-05508000 Sao Paulo - Brazil

^[2] Ctr Biotecnol I Butantan, Lab Microesferas & Lipossomas, BR-05503900 Sao Paulo - Brazil

^[3] Div Desenvolvimento Tecnol & Prod I Butantan, BR-05503900 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: MATERIALS; v. 2, n. 3, p. 765-775, SEP 2009.
• Citações Web of Science: 3

Resumo

During the W(1)/O phase (in the W(1)/O/W(2) 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 W(1)/O phase. A correlation was established between salts used in aqueous solutions and the changes in Dtxd solubility and conformation. The Dtxd a-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 CH(2)Cl(2) to begin its PLGA microencapsulation within ideal conditions. This was a technological breakthrough because a simple solution like salt addition avoided heterologous proteins usage. (AU)

• Processo FAPESP: 00/10970-7 - Microencapsulamento de macromoléculas em polímeros biodegradáveis
• Beneficiário: Maria Helena Bueno da Costa
• Modalidade de apoio: Auxílio à Pesquisa - Regular