Conformational stability and antibody response to the 18kDa heat-shock protein formulated into different vehicles.
Protein stability is one of the most important obstacles for successful formulation in the development of new-generation vaccines. Here, the 18kDa heat-shock protein (18kDa-hsp) was chemically modified though conjugation with bovine serum albumin or by esterification with N-hydroxysuccinimide ester of palmitic acid. The biologically active conformation of the protein was preserved after chemical modification. The immune responses to the recombinant 18kDa-hsp from Mycobacterium leprae were studied in different presentations: free, copolymerized with bovine serum albumin in aggregates (18kDa-hsp-BSA), and either surface linked to liposomes or entrapped into liposomes. Measuring the antibody production of immunized genetically selected mice has compared the adjuvant effects of liposomes and proteic copolymer. Among the two liposome preparations, the strongest response was obtained with the surface-exposed antigen-liposomes. The copolymer 18kDa-hsp-BSA conferred a high titer of antibody in injected mice, and persisted 70 d after immunization. This approach should prove very useful for designing more effective vaccines by using 18kDa-hsp as carrier protein.
Costa
M H
MH
Laboratório de Microesferas e lipossomos-Centro de Biotecnologia, Butantan, Brasil. mhbcosta@pasteur.fr
Sant'Anna
O A
OA
de Araujo
P S
PS
Sato
R A
RA
Quintilio
W
W
Silva
L V
LV
Matos
C R
CR
Raw
I
I
D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000818:Animals; D000917:Antibody Formation; D001426:Bacterial Proteins; D002417:Cattle; D004355:Drug Stability; D005260:Female; D006360:Heat-Shock Proteins / Q000008:administration & dosage* / Q000737:chemistry / Q000276:immunology*; D008081:Liposomes; D008297:Male; D051379:Mice; D009166:Mycobacterium leprae / Q000737:chemistry / Q000276:immunology; D014677:Pharmaceutical Vehicles; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology; D012710:Serum Albumin, Bovine / Q000008:administration & dosage; D018074:Vaccines, Conjugate / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology
D000276:Adjuvants, Immunologic; D001426:Bacterial Proteins; D006360:Heat-Shock Proteins; D008081:Liposomes; D014677:Pharmaceutical Vehicles; D011994:Recombinant Proteins; D018074:Vaccines, Conjugate; D014614:Vaccines, Synthetic; C081475:hsp18 protein, bacteria; D012710:Serum Albumin, Bovine