The use of protein structure/activity relationships in the rational design of stable particulate delivery systems

Autor(es)

M.H.B. Costa ^[1] ; W. Quintilio ^[2] ; O.A. Sant'Anna ^[3] ; A. Faljoni-Alário ^[4] ; P.S. de Araujo ^[5]

Afiliação do(s) autor(es)

^[1] Instituto Butantan. Centro de Biotecnologia. Laboratório de Microesferas e Lipossomos

^[2] Instituto Butantan. Centro de Biotecnologia. Laboratório de Microesferas e Lipossomos

^[3] Instituto Butantan. Laboratório de Imunogenética - Brasil

^[4] Universidade de São Paulo. Instituto de Química - Brasil

^[5] Universidade de São Paulo. Instituto de Química - Brasil

• Tipo de documento: Artigo Científico
• Fonte: Brazilian Journal of Medical and Biological Research; v. 35, n. 6, p. 727-730, 2002-06-00.
• Área do conhecimento: Ciências Biológicas - Biofísica
• Assunto(s): Biofísica Mycobacterium leprae Vacinas Epitopos

Resumo

The recombinant heat shock protein (18 kDa-hsp) from Mycobacterium leprae was studied as a T-epitope model for vaccine development. We present a structural analysis of the stability of recombinant 18 kDa-hsp during different processing steps. Circular dichroism and ELISA were used to monitor protein structure after thermal stress, lyophilization and chemical modification. We observed that the 18 kDa-hsp is extremely resistant to a wide range of temperatures (60% of activity is retained at 80ºC for 20 min). N-Acylation increased its ordered structure by 4% and decreased its ß-T1 structure by 2%. ELISA demonstrated that the native conformation of the 18 kDa-hsp was preserved after hydrophobic modification by acylation. The recombinant 18 kDa-hsp resists to a wide range of temperatures and chemical modifications without loss of its main characteristic, which is to be a source of T epitopes. This resistance is probably directly related to its lack of organization at the level of tertiary and secondary structures. (AU)

• Processo FAPESP: 00/14228-3 - Lipossomos e outros complexos lipídicos no estudo de vetorização de vacinas
• Beneficiário: Maria Helena Bueno da Costa
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 00/10970-7 - Microencapsulamento de macromoléculas em polímeros biodegradáveis
• Beneficiário: Maria Helena Bueno da Costa
• Modalidade de apoio: Auxílio à Pesquisa - Regular