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丁香实验推荐阅读
Gene Transfer and Drug Delivery by Electronic Pulse Delivery: A Nonviral Delivery Approach

Tremendous progress has been made in the understanding of human diseases and medical problems at a molecular level. This has led to the development of molecular medicine, that is, gene therapy and intracellular immunization. Among the major steps involved in the modification of target cells ...

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Effective Gene Transfer Using Viral Vectors Based on SV40

To date, virus-mediated gene transfer has focused principally on murine leukemia virus and adenovirus as delivery vehicles (1,2). It has become clear that these viral delivery systems are not adequate for the range of potential therapeutic needs, and other vehicles are being developed for t ...

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Strategies for Improving the Frequency and Assessment of Homologous Recombination

Gene therapy is a rapidly emerging field that holds much promise for the treatment of inherited disorders and acquired diseases. A major goal in most human clinical trials has been to produce long-term expression of therapeutic genes by integration of the DNA sequences encoding these genes in ...

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Rapid Generation of Isogenic Mammalian Cell Lines Expressing Recombinant Transgenes by Use of Cre Recombinase

Mammalian cell lines expressing a recombinant gene of interest are conventionally generated by random genomic integration of exogenous DNA containing the recombinant gene and a selectable marker. In contrast, site-specific recombination targets the exogenous DNA to a presele ...

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Site-Directed Alteration of Genomic DNA by Small-Fragment Homologous Replacement

The site-directed alteration of genomic sequences by homologous replacement (1,2), psoralen mutation (3), or site-directed repair-mediated correction (4)is a technology that can be used to achieve a number of different ends, including:the introduction of specific mutations into ...

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Mutation Correction by Homologous Recombination with an Adenovirus Vector

Gene targeting, the designed alteration of genomic information by homologous recombination, has provided a powerful means of genetic analysis of mammalian systems (1). Its wider application, especially to human gene therapy, is, however, hampered by its low level of efficiency. Only a ve ...

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Site-Specific Targeting of DNA Plasmids to Chromosome 19 Using AAV Cis and Trans Sequences

Adeno-associated virus (AAV) is of the genus Dependovirus and a member of the family Parvoviridae (1). AAV is unique, in that it requires co-infection of a second helper virus (i.e., adenovirus or herpesvirus) to undergo productive infection. In the absence of a helper virus, AAV will integrate pref ...

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What Is a B-Cell Epitope

The antigenicity of proteins resides in different types of antigenic determinants known as continuous and discontinuous epitopes, cryptotopes, neotopes, and mimotopes. All epitopes have fuzzy boundaries and can be identified only by their ability to bind to certain antibodies. An ...

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Structural Basis of AntibodyAntigen Interactions

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 ...

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A Solid-Phase Mutual Inhibition Assay with Labeled 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 ...

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Epitope Mapping of AntibodyAntigen Complexes by Nuclear Magnetic Resonance Spectroscopy

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 ...

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Epitope Mapping by Surface Plasmon Resonance

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 ...

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Epitope Mapping by Proteolysis of AntigenAntibody Complexes

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 ...

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Antibody Epitope Mapping Using SPOT Peptide Arrays

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 ...

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Linear B-Cell Epitope Mapping Using Enzyme-Linked Immunosorbent Assay for Libraries of Overlapping Synthetic Peptides

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 ...

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Peptide Microarrays for Profiling of Modification State-Specific Antibodies

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 ...

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Epitope Mapping Using Phage Display Peptide Libraries

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 ...

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Antibody Epitope Mapping Using De Novo Generated Synthetic Peptide Libraries

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 ...

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Antibody Specificity Profiling on Functional Protein Microarrays

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 ...

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Epitope Mapping Using Randomly Generated Peptide Libraries

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 ...

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