生物化学技术

Phage display can be used to generate human antibodies to virtually any antigen. This requires a number of crucial steps that are common to all molecular diversity technologies: the creation of diversity, coupling of phenotype (antibody protein) to genotype (antibody gene), selection, a ...

Central to the rapid growth in the study and application of recombinant antibodies in recent years is the ability to reliably and cost-effectively produce large quantities of functional protein. In contrast to glycosylated whole monoclonal antibodies (MAbs), functional derivati ...

Antibodies are becoming increasingly important for both basic research and clinical applications. With the completion of genome sequencing, antibodies are required for the global analysis and detection of every encoded protein. In therapeutics, a number of antibodies have been us ...

Since the advent of hybridoma technology, mammalian-cell culture has been employed for the expression and high-level production of monoclonal antibodies (MAbs). Recent adaptations in recombinant technology have developed the use of numerous prokaryotic and eukaryotic syst ...

Insect-cell expression systems provide a reliable and effective means for generating biologically active human antibodies. Two methods are available for this purpose. Most notably, the baculovirus expression system is a wellestablished tool for rapidly producing functional ...

The ability to mutate antibodies at the genetic level is a powerful tool in the antibody engineer’s toolbox. Specific properties of these molecules, such as stability, specificity, and in some cases catalysis, can be thoroughly studied using mutagenesis techniques. Harnessed to other te ...

Antibodies bind with great affinity and specificity to their target antigens, allowing them to be exploited in research, medicine, agriculture, and industry (1–3). It is estimated that more than 1,000 antibody-based pharmaceuticals are currently in development, and about 200 of these are ...

Phage display can be used to increase the affinity of antibodies more than 1,000-fold (1,2). The starting point is typically a specific antibody isolated from a phage antibody library (see Chapter 8). To accomplish affinity maturation (increased affinity), the sequence of the antibody is div ...

The immune response to an antigen can be considered to occur in two stages. In the initial stage, low-affinity antibodies are generated from an existing pool of the B-cell repertoires available at the time of immunization. In the second stage, which is driven by antigen stimulation, high-affinity ...

Since the advent of hybridoma technology a quarter century ago, a large number of murine monoclonal antibodies (MAbs) have been developed that are potentially useful clinical reagents against human infectious diseases and cancers. However, the clinical value of murine antibodies is l ...

The provision of an antibody or antibody fragment for use as an immunological reagent necessitates—at the very least—partial, but preferably total, purification of that antibody or antibody fragment from its source fluid. Numerous methods exist for this process, all designed to separa ...

The change in free energy for a reaction, ΔG, can be determined from the dissociation affinity constant, K D , since ΔG is equal to RTlnK D . The affinity constant provides no information on the rate of the reaction or the energy required to reach the transition state of the reaction (Fig. 1). Fig. 1.Energy vs reacti ...

High-affinity equilibrium binding constants for antibody-antigen interactions are inherently difficult to measure. Depending on the method used, difficulties arise from either the loss in signal-to-noise at the low concentrations that are necessary for measuring tight bind ...

The identification of antibody epitopes and their characterization at the amino acid level is extremely important. Understanding antibody specificity at the molecular level provides the key to optimizing their use as research or diagnostic tools as well as their application as ther ...

Fluorescence spectroscopy is a widely used technique for the characterization of molecular interactions in biological systems. It is highly sensitive, and allows measurements at low sample concentrations (down to 10-7 M). It is also available at a reasonably low equipment cost. These fe ...

Monoclonal antibodies (MAbs) are highly selective agents for delivering radiation to tumors both for in vivo imaging (radioimmunoscintigraphy, RIS) and for therapy (radioimmunotherapy, RIT). Antibody-based radiopharmaceuticals can be labeled with iodine nuclides (123I ...

Characterization of the antigen/antibody complex is an important tool in the rational design of antibody-based therapeutics. This can be achieved using several methods. Conventional methods for mapping an antibody-binding region on an antigen rely on synthetic polypeptide lib ...

Cocaine abuse has become a major health problem in recent decades, and the problem has been greatly accentuated by the appearance of crack cocaine, the more potent free base of the natural alkaloid. Cocaine’s primary molecular target is the dopamine reuptake transporter at the nucleus accumb ...

Recombinant immunotoxins are chimeric proteins composed of the Fv portion of a monoclonal antibody (MAb) fused to a portion of a toxin. The Fv replaces the cell-binding domain of the toxin and directs the toxin to cancer cells that express a target antigen. There are several features that make toxins ...

The explosion in genome sequencing, and the DNA array experiments based on those sequences, have provided a wealth of information on gene sequence, organization, and expression. This has led to a desire to carry out similarly broad experiments on all proteins encoded by a genome—the proteome. A ...