实验基础

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 ...

When attempting to establish libraries of immunoglobulins (Igs) from human subjects during the course of infection or other illness, a number of basic problems present themselves. First, the only source of lymphocytes that can be easily sampled is the peripheral blood in which the represen ...

The development of phage-display technology and the construction of huge libraries of antibody (Ab) fragments have provided an unlimited source of binders to virtually any antigen (Ag) (1). However, it is unlikely that the heavy (VH) and light (VL) chains of the Abs obtained from these libraries r ...

Recombinant antibody (Ab) libraries have been constructed from a wide range of B-lymphocyte sources using a number of different approaches. Sizes of the libraries that have been produced vary considerably, from small libraries of 106 up to large libraries1010. Often an Ab with the desired sp ...

Food, water, pharmaceutical, and medical microbiology ideally need methods of detecting, identifying, and quantifying microorganisms that can give results within a few hours. The many days needed for conventional enrichment, plating, and biochemical/serological methods to g ...

Many areas of analytical microbiology that deal with biological materials (e.g., medicine and the food, water, and pharmaceutical industries) need methods of detecting and identifying microorganisms that can give results in a few hours rather than the many days required for convention ...

To understand the principle of an immunoassay, one must be familiar with the antibody-antigen relationship. An antigen is a molecule that, when injected into an animal, will elicit an antibody response. Antigens may be proteins, polypeptides, or carbohydrates (glycolipids or glycopro ...

The isolation of genomic DNA from a microorganism generally comprises three stages: cultivation of the cells, disruption to release cell contents, and chemical purification of the DNA. Two widely used methods for the preparation of bacterial DNA are those described by Marmur (1) and Kirby (2), ...