抗体抗原实验

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 ...

The use of synthetic peptide immunogens as a means to generate specific immunological reagents for a variety of purposes has increased markedly in recent years. In this chapter, we outline some of the salient factors to be considered when designing peptide immunogens and describe basic meth ...

Hybridomas are derived from the fusion of spleen and myeloma cells and produce monoclonal antibodies. Each hybridoma cell line produces an antibody with a unique specificity allowing the production of highly defined reagents that can be used in many branches of immunochemistry.

Hybridomas can be grown in a number of ways to produce stocks of monoclonal antibodies. Smaller volumes may be produced in static flask culture, but if larger amounts are required, a bioreactor may be used.