Comparison of three ELISA protocols to measure antibody responses elicited against serogroup C meningococcal polysaccharide in mouse, monkey and human sera

• Published in: Journal of microbiological methods 2006-04, Vol.65 (1), p.135-143
• Main Authors: Guirola, María, Carmenate, Tania, Menéndez, Tamara, Álvarez, Anabel, González, Sonia, Guillén, Gerardo
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Bacterial - biosynthesis; Antibodies, Bacterial - blood; Bacterial Capsules - immunology; Bacteriology; Biological and medical sciences; Capsular polysaccharide; Cercopithecus aethiops; ELISA; Enzyme-Linked Immunosorbent Assay - methods; Female; Fundamental and applied biological sciences. Psychology; Genetics; Humans; Immune response; Meningitis, Meningococcal - immunology; Meningitis, Meningococcal - microbiology; Meningitis, Meningococcal - pathology; Meningococcal Vaccines - immunology; Mice; Mice, Inbred BALB C; Microbiology; Neisseria meningitidis; Neisseria meningitidis, Serogroup C - immunology; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains; Polysaccharides, Bacterial - immunology; Statistics, Nonparametric; Vaccines, Conjugate - immunology
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2005.06.015

In this study we compared the following ELISA protocols to measure antibody levels against serogroup C meningococcal polysaccharide: a traditional protocol using poly- l-Lysine mixed with the polysaccharide as coating antigen, a second protocol coating with a mixture of methylated human serum albumin with the C polysaccharide, a modified protocol coating with derivatized polysaccharide and a modification to the last one, specifically without adding ammonium thiocyanate to the sample buffer. Serum bactericidal activity of mouse, monkey and human sera were measured and correlation coefficients were calculated. For all serum types the modified ELISA protocol showed the highest correlation coefficients while the traditional protocol showed the lower ones. We demonstrated that the traditional protocol measures non-specific antibodies to the C polysaccharide, because no differences were detected between pre-immune and post-immune human sera ( P > 0.05), while the modified protocol detected the highest difference ( P < 0.01).