基因表达差异显示实验

Antibody molecules can be regarded as products of a protein engineering system for the generation of a virtually unlimited repertoire of complementary molecular surfaces. This extreme structural heterogeneity is required for recognition of the nearly infinite array of antigen ...

A widely applicable method for the determination of the epitope specificities of a large number of monoclonal antibodies (MAbs) is presented. The method is based on the solid-phase mutual inhibition assay using 96-well plates coated with the respective MAbs, competitor MAbs, biotinyla ...

Nuclear magnetic resonance (NMR) is a very powerful tool for determining the boundaries of peptide epitopes recognized by antibodies. NMR can be used to study antibodies in complexes that exhibit a wide range of binding affinities from very weak and transient to very tight. Choice of the specif ...

Biacore™ systems (Biacore AB) enable label-free detection of molecular interactions in real time using surface plasmon resonance detection. Epitope mapping of antibodies is usually performed in a pairwise fashion where one antibody is used to capture the antigen from solution and the b ...

The ability to accurately characterize an epitope on an antigen is essential to understand the pathogenesis of an infectious material, and for the design and development of drugs and vaccines. Emergence of a new contagious microbial or viral variant necessitates the need for robust identi ...

Information at the amino acid level about the epitopes of proteins recognized by antibodies or antibody fragments is important for their use as biological and diagnostic tools, therapeutic molecules, and for understanding molecular recognition events in general. The use of chemica ...

The aim of this chapter is to provide a strategy for mapping linear antibody epitopes of protein antigens in order to discover candidates for vaccines or diagnostic tests. A set of overlapping peptides was designed and synthesised based upon a known amino acid sequence of the target protein, viru ...

The reversible phosphorylation of serine, threonine, and tyrosine residues is one of the most important intracellular post-translational modifications regulating enzymatic activities and protein/protein interaction in eukaryotic cells. Tools for determining phos ...

Phage libraries displaying millions of peptides with randomized sequences are extremely useful tools for mapping antibody epitopes. In many cases, antibodies are able to select peptides with reasonable affinity for their combining sites (paratopes) from these libraries. Ideal ...

Identification of antibody binding peptides may be based on the primary structure of the protein antigens used to raise the antibodies (knowledge- or sequence-based approach). This involves scanning the entire sequence of the antigen with overlapping peptides (peptide scan), which a ...

Antibodies represent the end product of an exquisitely complex biological process including recombination, somatic hypermutation, affinity maturation, and self-tolerance, culminating in binding reagents directed against a vast repertoire of antigens. The resultant hi ...

Characterizing the immune response towards a pathogen is of high interest for vaccine development and diagnosis. However, the characterization of disease-related antigen–antibody interactions is of enormous complexity. Here, we present a method comprising binding studies of ...

Polyclonal antibodies raised against full-length antigens are often used for localization experiments. Exact knowledge of epitopes in the antigen recognized by the antiserum is important if the target antigen belongs to a large family of proteins which are highly conserved. We have sh ...

High density peptide microarray technologies can be applied in experimental medicine in general and in clinical immunology in particular. Laboratory diagnostics of autoimmune diseases strongly rely on screening human sera for antibodies against known autoantigens. These as ...

With the evolution of peptide synthesis techniques and microarray technology, it is now possible to map and characterize allergenic epitopes on a microarray platform. The peptide microarray-based immunoassay allows simultaneous analysis of thousands of target peptides using ...

Successful adaptation of microarray technology for high-throughput screening of proteins requires a large number of purified recombinant proteins, e.g., antibodies for use as capture molecules. Phage surface display technology has been used for the surface expression of protei ...

With the advance of whole genome sequencing for an increasing number of organisms, it becomes clear that many proteins exist in multiple forms whose overall structures are similar despite subtle sequence and functional differences. Although the biological significance may not be kno ...

We will describe a procedure to map epitopes on a protein against monoclonal IgGs. In this procedure, we amplified and mutagenized the entire or a part of the protein. Then, a DNA region encoding the protein was cut out by a restriction enzyme and ligated into a lambda-gt11 expression vector to construct a ...

The prediction of B-cell epitopes is desirable for designing peptide-based vaccines, or generating antibodies especially if the purified protein is difficult to obtain and immunization has to be performed with protein-derived synthetic peptides. A number of freely available too ...

Phage-display has become a method of choice for epitope mapping and has been successfully used in numerous published studies. Although the inaugural studies were all done with random peptide libraries (see Chapter “Epitope Mapping Using Phage Display Peptide Libraries”), gene- or geno ...