Structural characterisation, stability and antibody recognition of chimeric NHBA-GNA1030: An investigational vaccine component against Neisseria meningitidis

• Published in: Vaccine Vol. 30; no. 7; pp. 1330 - 1342
• Main Authors: Martino, Angela, Magagnoli, Claudia, De Conciliis, Giuseppe, D’Ascenzi, Sandro, Forster, Mark J, Allen, Lauren, Brookes, Charlotte, Taylor, Stephen, Bai, Xilian, Findlow, Jamie, Feavers, Ian M, Rodger, Alison, Bolgiano, Barbara
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 08.02.2012
• Subjects: Adhesins, Bacterial - genetics; Adhesins, Bacterial - immunology; Adhesins, Bacterial - metabolism; Allergy and Immunology; Animals; Antibodies, Bacterial - biosynthesis; Antibodies, Bacterial - immunology; antigens; Antigens, Bacterial - genetics; Antigens, Bacterial - immunology; Antigens, Bacterial - metabolism; chimerism; chromatography; Circular Dichroism; electrophoresis; Equilibrium unfolding; fluorescence emission spectroscopy; Fusion protein; Heparin - immunology; Heparin - metabolism; Heparin binding antigen; Hot Temperature; immune response; Immunoglobulin G - biosynthesis; Immunoglobulin G - immunology; Meningococcal Vaccines - genetics; Meningococcal Vaccines - immunology; Mice; Mice, Inbred C57BL; Models, Molecular; Neisseria meningitidis; Neisseria meningitidis, Serogroup B - immunology; Protein Folding; Protein Stability; Protein Structure, Secondary; Proteolysis; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - immunology; serotypes; Spectrometry, Fluorescence; Spectrometry, Mass, Electrospray Ionization; tryptophan; vaccines; CDA; fusion protein (NHBA-FP); porin A; size-exclusion chromatography; PorA; λ ex; circular dichroism; neisserial heparin-binding antigen; bicinchoninic acid; ESI-Q-TOF; SBA; serum bactericidal antibody; NHBA-GNA1030; Rh; fluorescence emission wavelength maximum; Q-ELS; Fusion protein; NHBA; GuHCl; Neisseria meningitidis serogroup B; F max; BCA; CD; MALLS; multiangle laser light scattering; M w; d n/d c; complement deposition assay; Heparin binding antigen; MenB; Neisseria meningitidis; polydispersity; weight average molar mass; excitation wavelength; hydrodynamic radius; guanidine hydrochloride; SEC; differential index of refraction; quasi elastic light scattering; Equilibrium unfolding; RI; genome-derived neisserial antigen; GNA; refractive index; electrospray ionisation-quadrupole time of flight; M w/ M n; Mw; Mw/Mn; dn/dc; λex; Circular dichroism; Fmax
• ISSN: 0264-410X; 1873-2518
• DOI: 10.1016/j.vaccine.2011.12.066

Highlights ► We describe the biophysical characteristics of the NHBA-GNA1030 vaccine antigen. ► The fusion protein has folded structure in the NHBA C-terminal domain and in GNA1030. ► Thermal stress causes little change in protein secondary structure. ► Exposure at 37 °C for up to 15 days results in protein degradation and aggregation. ► Maintenance of the intact, non-fragmented state preserves functional immunogenicity.