微生物学技术

Several approaches have been used to identify antibody (Ab)-defined human tumor-associated antigens (TAA) that can be used for passive and/or active specific immunotherapy of malignant diseases. For example, hybridoma technology has been successfully used to identify TAA recog ...

Molecular evolution approaches to developing molecules with characteristics particularly suited for specific applications have become important tools in biomedicine and biotechnology. Not only is it possible to identify molecules with specificities that cannot easily ...

Cell and/or tissue-specific phage antibodies (Abs) are usually generated using single-cell suspensions as a starting material for selection. However, isolation of Abs specific for certain cells and/or tissue components may be hampered because the natural architecture of the tiss ...

Antibodies (Abs) displaying an agonist or antagonist activity are powerful tools for mimicking or blocking physiological functions in the cell. A number of applications of Abs in diagnosis and therapy require multivalent reagents, either because biological activity depends on the ...

The potential of Fvs as magic bullets for targeting therapeutic drugs or imaging agents is well-documented. However, the conversion of whole antibodies (Abs) into Fvs (scFvs or dsFvs) is often associated with a drastic reduction in antigen (Ag)-binding affinity. For efficient use of Fvs as tar ...

A two-step strategy for changing the specificity of antibodies (Abs) is presented, which we have used to change the specificity of an Ab from 11-deoxycortisol (11-DOC) to cortisol (CS). Two kinds of in vitro mutagenesis are utilized in this protocol: first, mutations are introduced at restricted ...

Despite the power of antibody (Ab) phage-display technology, a problem which can be commonly encountered is the recovery of Abs of low affinity for the antigen (Ag) of interest. Two general strategies can be applied to increase affinity: mutations can be scattered randomly throughout the gene ...

The use of antibody (Ab) molecules and their fragments in research, diagnosis, and therapy has prompted the development of methods to improve their affinity and stability to increase their expression levels and to change or improve their specificity. This is easier to carry out on Ab fragments ( ...

Combinatorial libraries and selection of variants from such libraries have proven to be a successful approach for identifying molecules with novel or improved properties. The importance of antibody (Ab) molecules in basic and applied research, as well as the extensive knowledge of how t ...

In the past few years, some of the limitations of monoclonal antibodies (MAbs) as therapeutic agents have been addressed by genetic engineering. Such an approach is particularly suitable because of the domain structure of the Ab molecule, where functional domains carrying antigen (Ag)-b ...

Fab libraries, in which light-chain (LC) and heavy-chain (HC) variableregion genes are cloned into a phagemid vector and subsequently displayed on the surface of the filamentous phage particle, have been widely used for the isolation of antibodies (Abs) with specificity for haptens, fore ...

Since the advent of hybridoma technology 25 years ago, monoclonal antibodies (Abs) have revolutionized many aspects of biological research and health care. After some initial setbacks, Abs are also beginning to make an impact as therapeutic agents in the clinic (1). In the last decade, novel se ...

Molecular techniques for inhibiting the expression of specific genes represent a highly refined approach to the analysis and manipulation of microbial and cellular pathways. The specific and high affinity binding properties of antibodies (Abs), combined with their ability to be st ...

The use of Saccharomyces cerevisiae as a production host for heterologous proteins has several advantages compared to the use of a bacterial host. First, S. cerevisiae has proven to be a productive host capable of expressing many heterologous proteins at high levels. Second, because S. cerevi ...

scFvs have considerable therapeutic potential against antigens (Ags) involved in disease processes (1 ,2), either as proteins synthesized ex vivo for passive administration, or introduced by gene therapy for in vivo expression. Their small size confers many pharmacological adva ...

Since the generation of the first human antibodies (Abs) by phage display (1,2), technology has evolved to allow the creation of large, nonimmunized fully human scFv repertoires that yield Abs with comparable affinities to those obtained using hybridoma technology (3,4). Using a variety of s ...

The technologies described in this volume enable the isolation of recombi- nant antibodies (Abs) from libraries of variable regions (Fvs) or Fabs displayed on the surface of filamentous phage by fusion to a structural protein (1 -7). Production of selected Abs in sufficient quantity for furt ...

In recent years, a number of single-pot antibody (Ab) libraries have been described, which permit the rapid isolation of high-affinity Abs against large panels of antigens (Ags). Na�ve libraries have been generated by tapping the natural primary (unselected) immune repertoire via cloni ...

The production of monoclonal antibodies (MAb) through the immortalization of B lymphocytes has generally had little impact beyond human and murine immunology. This can be explained by the lack of appropriate myeloma lines or transforming viruses for species outside this select group a ...

Polyclonal antibody libraries (PCALs) are standardized mixtures of antibodies (Abs) specific for an antigen (Ag) or multi-Ag target (a poly-Ag). As the immunoglobulin (Ig) genes are cloned, the mixtures can be perpetuated, amplified, and modified as desired. Poly-Ags of special interest ...