title: The use of protein structure/activity relationships in the rational design of stable particulate delivery systems.
abstract: 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 80 degrees C for 20 min). N-Acylation increased its ordered structure by 4% and decreased its beta-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.
authors.affiliation: Laboratório de Microesferas e Lipossomos, Centro de Biotecnologia, Instituto Butantan, São Paulo, SP, Brasil. bdacosta@edu.usp.br
authors.forename: M H B
authors.initials: MH
authors.lastname: Costa
authors.forename: W
authors.initials: W
authors.lastname: Quintilio
authors.forename: O A
authors.initials: OA
authors.lastname: Sant'Anna
authors.forename: A
authors.initials: A
authors.lastname: Faljoni-Alário
authors.forename: P S
authors.initials: PS
authors.lastname: de Araujo
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D001428:Bacterial Vaccines / Q000737:chemistry; D004355:Drug Stability; D004797:Enzyme-Linked Immunosorbent Assay; D006360:Heat-Shock Proteins / Q000032:analysis*; D009166:Mycobacterium leprae / Q000737:chemistry*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D013329:Structure-Activity Relationship; D013696:Temperature
chemical_list: C067864:18 kDa protein, Mycobacterium leprae; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins