免疫学技术

The evaluation of meningococcal vaccines in humans is a challenging task. Issues of safety, and benefit to the individual patient and to the community may raise difficult ethical problems. The inherent variability of human responses, the rarity of clinical disease, age-dependence in the i ...

Meningococcus is an aerobic, fastidious, Gram-negative diplococcus that is found only in humans. Its cell wall has a cytoplasmic membrane, a peptidoglycan layer, and an outer membrane. Meningococci isolated from the bloodstream or the spinal fluid are almost always encapsulated. The ca ...

For the most part, the relationship between the pathogen, Neisseria meningitidis, and humans is uneventful. Colonization of the human nasopharynx at various times during life is an almost universal experience but clinically overt disease is unusual except during epidemics. This ove ...

The use of bacterial capsular polysaccharides as immunoprophylactic agents in human diseases caused by encapsulated bacteria is now firmly established (1). However, despite their many advantages, they do have serious limitations. First, they induce an inadequate immune response ...

Serogroup B Neisseria meningitidis remains a major world health problem and currently there is no fully efficacious vaccine available. The poor immunogenicity of the group B meningococcal polysaccharide both in adults and infants prevents the formulation of a comprehensive poly ...

The major outer membrane proteins (OMPs) from Neisseria meningitidis, which are expressed at high levels, are subdivided in five classes based on molecular weight (1,2) (see Table 1). Table 1 Major Meningococcal Outer-Membrane Proteins

Alternative strategies exist for prevention of group B Neisseria meningitidis (meningococcal) disease through vaccination (see Chapters 5, 8, 13, 14 in this volume). However, the most promising approach to date has been the use of outer-membrane vesicle (OMV) vaccines for induction of ba ...

An understanding of the molecular basis for the recognition of outer-membrane proteins (OMPs) by antibody is an important goal in the development of a more rational approach to vaccine design. X-ray crystallography has been outstandingly successful in delineating the detailed chemi ...

To obtain iron, a necessary nutrient, meningococci and several other human and veterinary pathogens have iron-acquisition systems, which are expressed in vivo during infection. One target of iron-acquisition systems is transferrin (Tf), which is the major glycoprotein responsib ...

The isothiocyanate derivative of the fluorophore, fluorescein or rhodamine, is coupled to the amino groups of IgG antibody in a one-step procedure and excess label is removed by gel filtration.

The detection of virus in samples using electron microscopy can be enhanced by the use of specific antibodies to trap particles. Antibodies can further be used to label immobilized particles on grids to aid their identification.

Antibodies can be conjugated to biotin by a number of chemical means. They can then be used in immunochemical procedures in conjunction with secondary reagents coupled to biotin-binding protein proteins such as avidin.

Competitive enzyme-linked immunosorbent assay provides an alternative to dual-antibody sandwich enzyme-linked immunosorbent assay and is widely used for the measurement of substances in biological liquids.

Antibodies can be used to measures levels of molecules in solution using the dual-antibody sandwich enzyme-linked immunosorbent assay. Two antibodies are used that become bridged in the presence of the substance of interest, leading to the production of a colored product by a reporter enzy ...

Immunoblotting techniques use antibodies (or other specific ligands in related techniques) to identify target proteins among a number of unrelated protein species. They involve identification of protein target via antigen-antibody (or protein-ligand) specific reactions. ...

Immunoaffinity purification is a highly specific, reversible technique that has the potential to be used for the one-step isolation of an analyte from many complex matrices. Application of an immunoaffinity column to isolate and concentrate an analyte may decrease the amount of solvent ...

Antibody capture of viruses can be used as a preparatory step in nucleic acid amplification techniques. Immunocapture of virus particles can be used to streamline and/or optimize the concentration, purification and specificity requirements of polymerase chain reaction assays.

Antibodies reacting to specific cell surface markers can be bound to magnetic beads and used to specifically capture cells exhibiting the marker. This approach allows the selective enrichment of specific cell subpopulations and as the methods are amenable to use in blood, tissue fluids and ...

Cell populations can be enriched using a number of physicochemical methods based on characteristics, such as density, size, and volume. Centrifugation using density gradients is typically applied to isolate populations with similar characteristics. Cells obtained using these ...

The generation of polyclonal antibodies to an antigen of interest is an important technique applicable to many areas of biological research. In this chapter, we describe a basic immunization procedure designed to generate polyclonal antisera in rabbits and two methods that are commonly ...