Crystallization and preliminary X-ray diffraction analysis of antigen-binding fragments which are specific for antigenic conformations of sialic acid homopolymers.
Meningococcal meningitis is a severe childhood disease which often results in significant disability or death. Two major etiological agents of meningitis are the group B meningococci and capsular type K1 E. coli. The virulence of these organisms is attributable to structural mimicry between their common alpha(2-8)-polysialic acid capsular polysaccharide and human tissue antigens, which allows the bacteria to evade immune surveillance. There is currently no effective vaccine to protect against this infection. It has been demonstrated that the capsular polysaccharide of the bacteria can adopt a unique 'antigenic conformation'. This antigenic conformation has formed the basis for the development of an N-propionylated polysialic acid vaccine. Immunization trials in mice with this vaccine show the production of two groups of antibodies, of which only N-propionylated polysialic acid-specific were protective. Knowledge of the structure of the antigen-binding site which recognizes the protective epitope is essential to determining the antigenic conformation of the polysaccharides, and is a critical aspect in understanding and improving the action of potential vaccines. The antigen-binding fragments (Fab) of one protective (13D9) and one non-protective (6B9) monoclonal antibody specific for the capsular polysaccharides of group B meningococci have been crystallized and have undergone preliminary X-ray diffraction analysis. Both crystals are observed to scatter X-rays to approximately 1.7 A resolution at the A1 station at the Cornell High-Energy Synchrotron Source. 13D9 has an orthorhombic unit cell with a = 41.8, b = 102.3, c = 134.7 A, with space group P212121. Fab 6B9 has an orthorhombic unit cell with a = 89.6, b = 132.0 and c = 36.9 A, with space group P21212.
Department of Biochemistry, University of Ottawa, 451 Smyth, Ottawa, Ontario K1H 8M5, Canada.
S I
SI
Patenaude
S M
SM
Vijay
Q L
QL
Yang
H J
HJ
Jennings
S V
SV
Evans
D000818:Animals; D000907:Antibodies, Bacterial / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D000942:Antigens, Bacterial / Q000737:chemistry / Q000276:immunology*; D016667:Bacterial Capsules; D001666:Binding Sites, Antibody; D002236:Carbohydrate Conformation; D003460:Crystallization; D018360:Crystallography, X-Ray; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008814:Mice, Inbred NZB; D019158:N-Acetylneuraminic Acid / Q000737:chemistry / Q000276:immunology*; D009345:Neisseria meningitidis / Q000737:chemistry / Q000276:immunology*; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000276:immunology*; D011487:Protein Conformation
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D011135:Polysaccharides, Bacterial; C039455:capsular polysaccharide, meningococcal group B; D019158:N-Acetylneuraminic Acid