免疫学技术

The construction of very large combinatorial libraries whereby antigen-binding functional domains of antibodies (Abs) are cloned and displayed on the surface of bacteriophage is proving to be a powerful technology in providing reagents for immunotherapy. Combinations of rand ...

Phage display involves expression of a large library of diverse molecules as fusion proteins on the surface of filamentous bacteriophage. In this form, they may be subjected to biological or molecular selection to isolate molecules with the desired binding properties (1) (see review in ref. ...

Apart from cell-cell contacts, signal transduction mechanisms active during immune defense are mainly regulated by cytokines. They accomplish their pleiotropic, paracrine, and/or autocrine biological activities as a result of the contact with a membrane receptor followed by si ...

Monitoring cytokine responses to various stimuli or determining the expression pattern of cytokine mRNAs requires sensitive technologies for cytokine mRNA quantitation where there is a limited quantity of material, for example, when working with biopsies. Owing to their amplif ...

Chemokines, i.e., low-molecular-weight chemotactic cytokines, are generally produced in very small amounts. Hence, liters of conditioned medium from primary cells or cell lines are necessary as start material to obtain microgram amounts of natural chemokine (see Note 1). A substanti ...

In situ hybridization (ISH) involves hybridization of labeled nucleic acid probes with the target mRNA. As cytokines are soluble mediators exported from the cells in which they are produced, ISH is a powerful technique that can be used to identify the exact identity and location of the cells that p ...

Affinity chromatography as it is known today, namely, the concept and the immense power of biorecognition as a means of purification, was introduced in 1968 by Cuatracasas, Wilchek, and Anfinsen (1). This technique is used in 60% of all purification protocols (2). Almost any given biomolecule that ...

Crystallography is the most powerful method for determining the three-dimensional structure of complicated biological molecules. As the technology has improved, larger and more complicated structures involving previously unobtainable proteins have become available. ...

Cytokines trigger growth factor responses through interaction with cell surface receptors. Determining the quantitative properties of cytokine-receptor interactions can help lead to mechanistic understanding of receptor activation as well as strategies to inhibit the i ...

βlactam resistance in clinical isolates of Streptococcus pneumoniae arises by only one route, the reduction of the affinity of the penicillin-binding proteins (PBPs) for βlactams. The pneumococcus possesses five high molecular weight PBPs (PBP1A, 1B, 2A, 2B, and 2X) which are involved in t ...

Fluoroquinolones act by inhibiting the essential type II topoisomerases, DNA gyrase and topoisomerase IV (1). DNA gyrase is a tetramer composed of 2 A subunits and 2 B-subunits encoded for by gyrA and gyrB, respectively. This enzyme is the only enzyme known that is capable of negatively supercoil ...

Penicillin-resistance in Streptococcus pneumoniae is mediated by altered penicillin-target enzymes, the penicillin-binding proteins or PBPs. PBPs interact withβ-lactam antibiotics by forming an active penicilloyl-PBP complex via an active site serine. This complex is en ...

The ability to share genetic information with other bacteria represents one of the most important adaptive mechanisms available to bacteria pathogenic for humans. The exchange of many different types of genetic information appears to occur frequently and exchange of determinants ...

Monoclonal antibodies (MAbs) are essential reagents for the isolation, identification, and cellular localization of specific gene products and for aiding in the determination of their macromolecular structure. They can also help in identifying the function of the protein. Altho ...

Monoclonal antibodies (MAbs) have traditionally been produced in quantity by transfer of the hybridomas to the peritoneal cavity of mice and subsequent recovery of MAb from the ascitic fluid. Legislation in many countries, however, now limits the use of animals for such purposes. Although ...

The production of large quantities of monoclonal antibodies (MAbs) of predetermined specificity has been rendered possible by the pioneering work of K�hler and Milstem (1) These workers have shown that lymphocytes can be immortalized and subsequently cultured after so1atic fusion ...

An increasing problem in cell and molecular biology is the preparation of antibodies specific to proteins that are present in minute quantities within cells or tissues. With the advent of recombinant DNA technology, it is now often possible to deduce the primary amino acid sequence of a polypep ...

The great analytical power of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) makes it one of the most effective tools of protein chemistry and molecular biology. In the past, there have been many attempts to convert the technique from analytical to preparative s ...

Glycosylation is a major posttranslational modification that produces heterogeneity in a protein and results in a group of structurally different species called glycoforms. Protein-associated oligosaccharides are large, structurally diverse molecules, and the biolog ...

Until recently, the role of antibodies in graft failure has been hampered by poor methods of defining specificity. Development of solid phase assays using purified major histocompatibility complex (MHC) molecules has greatly advanced our ability to monitor anti-human leukocyte an ...