免疫学技术

Various methods have been devised to detect antibody-producing cells in culture and also determine the isotype of the immunoglobulin produced. We have found the ELISPOT assay to be the method of choice for detecting human monoclonal antibodies (MAbs) produced by heterohybridomas (see ...

One of the easiest and most sensitive methods to screen for, to quantify, and to determine the quality of antibodies in the serum of mice, the spleens of which may be used for fusions, is by solid-phase enzyme-linked immunoassays (EIA) (1,2). In these procedures, the wells of 96-well plates or strip plates are ...

Immunohistochemistry (IHC) or immunocytochemistry is a method for the localization of specific antigens in tissues or cells using antibody probes. Because it provides detailed and specific information on the expression of cell markers, the IHC technique has wide application in the i ...

Immunoelectron microscopy (IEM) techniques, usually in conjunction with light microscopy techniques, are used for the localization of antigenic molecules that may be employed for vaccine development, organelle characterization, or other biologically relevant applica ...

Protein blotting involves the immobilization of proteins on a support medium. The most usual format is the electrophoretic transfer of proteins resolved on a sodium dodecyl sulfate (SDS)-polyacrylamide gel to a membrane, which is known as Western blotting (1). Polyacrylamide gel elect ...

Recent advances in the design of flow cytometers and the development of associated software have facilitated the use of flow cytometry as a primary method for identifying and characterizing monoclonal antibodies (MAbs) reactive with molecules expressed on one or more lineages of leuk ...

Mammalian cells from every tissue have many different types of proteins on their extracellular membranes that maintain their functional integrity. These molecules participate in communication, in metabolism, in locomotion, or in adherence. Antibodies developed against the ...

Until recently, the primary method available for the production of large quantities of monoclonal antibodies (MAbs) has been growing hybridomas in syngeneic or nude mice. This has presented a number of problems. The first has been the introduction of contaminating proteins. The second has ...

Thiophilic adsorption chromatography was first introduced by Porath and Belew (1) and was described as a one-step method for purification of monoclonal antibodies (MAbs) (2). As such, the method seems very attractive. Moreover, it operates with lyotropic salts, which are most agreeable r ...

Antibodies of the IgM class are elaborated by many common murine hybridoma cell lines and often define important antigens. In contrast to IgG antibodies, however, IgM molecules exhibit little or no affinity for bacterial Ig binding proteins, such as protein A and protein G, which are almost univ ...

In the past, isolation of a pure protein demanded many hours to develop a purification protocol, which included one or more cycles of time-consuming chromatography, only to have a final product that was enriched rather than purified. The development of a purification procedure had to be repeat ...

Until recently, the primary methods available for preparing immunoaffinity columns with both polyclonal and monoclonal antibodies (MAbs) have required purification of the antibodies from serum, ascites, or tissue-culture medium prior to coupling to the affinity matrix beads or ...

The avidin-biotin bond is the strongest known biological interaction between a ligand and a protein (K d = 1.3 � 10−15 M at pH 5) (1). The affinity is so high that the avidin-biotin complex is extremely resistant to any type of denaturing agent (2). Biotin (Fig. 1) is a small, hydrophobic molecule that functions as a c ...

Immunofluorescence analysis has been greatly aided by the use of monoclonal antibodies (MAbs) modified by derivatization with fluorescent labels (1). Improvements of known fluorophores and development of new ones with a broader range of colors have paralleled the production of new d ...

As an outgrowth of the experience obtained from the chemical modification of proteins, a series of techniques to crosslink different proteins chemically has been developed in the last two decades. The conjugation of enzymes to antibodies, particularly to monoclonal antibodies (MAbs ...

Sorting heterogeneous populations of cells into specific subpopulations has been greatly facilitated by the advent of monoclonal antibodies (MAbs) in combination with magnetic particles (1–35). MAbs provide the specificity necessary to distinguish one cellular subpopula ...

The use of magnetic particles in combination with monoclonal antibodies (MAbs) has greatly simplified sorting heterogeneous populations of cells into specific subpopulations (1–35). MAbs which recognize particular cell surface markers provide the specificity necessary to ...

In vitro immunization involves the exposure of spleen cells to antigen in tissue culture rather than the antigenic stimulation of spleen cells via immunization of mice. The production of monoclonal antibodies (MAbs) to highly conserved molecules, such as enzymes (1,2), is possible using in ...

Efforts to refine the methods of producing monoclonal antibodies (MAbs) of known specificity (1) have revealed there are many variables that affect the growth of hybridomas generated by the fusion of myeloma cell lines with spleen cells (reviewed in refs. 2 and 3). These include the cell-cycle st ...

In many circumstances, it is advantageous to have a continuous source of human antibody of a given specificity and immunoglobulin isotype. Reliance on human volunteers as a source of such antibody is problematic. Therefore, it has been a goal of investigators to establish immortal cell lines t ...