Antibodies antigen injection

To test DC-specific targeting, model antigens such as hen egg lysozyme (HEL) and ovalbumin (OVA) were covalently linked to anti-DEC-205 antibodies and injected into mice. This resulted in antigen loading of the lymph node DCs,... 

To produce monoclonal antibodies, the first antigen injection in mice is followed a few weeks later by a booster of the same antigen. When polyclonal antibodies are detected in the serum of mice a few days after the booster injection,... 

Very few immunosensors are commercially available. The commercial immunosensors are either the detector or bioanalyzer types. The PZ 106 immunosensor from Universal Sensors Inc. (New Orleans, LA) has been used as a detector to measure antibody-antigen reaction. Ohmicron (Newtown, PA) developed a series of pesticide immuno-bioanalyzers that have been used in field tests. Pharmacia Biosensor USA (Piscataway, NJ) recently introduced BIAcore immunodetection system. A combination of a unique flow injection device and surface plasmon resonance (SPR) detection technique provides a real time analysis. A carboxylmethyldextran layer added to plasmon generating gold film is a hydrophobic, activatable, and flexible polymer that provides high antibody and low non-specific bindings. System demonstration at the Institute of Food Technologists (IFT) 1994 meeting in Atlanta drew attention of food scientists. It should easily be adapted for food protein characterization. 

Immunization procedures vary and are dependent on type of antigen to be used, duration of immunization process, and the amounts of immune product needed. The antigen suspension may be administered intravenously, intramuscularly, or subcutaneously. The amount of antigen injected can range from 1 to 200 mg. The quantity is determined by the availability of and the potency of the antigen. The time schedule also varies. Protocols for the three types of immunizations used to produce anti-carbohydrate antibodies are recorded in the following. 

In general, drugs have molecular weights which are too low for them to act as antigens, i.e. to produce antibodies when injected into an animal. It is necessary, therefore, to couple the drug to a larger molecule which is usually a protein. The most commonly used protein is albumin (molecular weight approximately 70 000), taken from a species other than that in which the antibody is to be raised. 

A detailed discussion of antibody production and its control is beyond our purpose. However, minimal appreciation of the events leading to antibody production and a few definitions are necessary if immunochemical techniques are to be used knowledgeably. An antigen may be defined as any compound that (1) can stimulate production of antibodies when injected into a test animal and (2) reacts specifically with the antibodies produced. Both parts of this definition are necessary to distinguish antigens from haptens (see Figure 8-3), which are small molecules that... 

Antigen Injected (mg) Average Intrinsic Association Constants for Binding of c-DNP-Lysine to Anti-DNP Antibodies ... 

Procedures for preparation of the immunizing antigen, injection schedules, and assays for antibody formation are similar to those used for helical polynucleotide-MBSA antigens and are discussed in detail in the following section and in Vol. 12B [174] of this series. 

On the basis of an enzyme thermistor, Mattiasson et al. (1977) developed one of the first immunosensors. Immobilized antibodies against albumin are placed in a column and set into an ET. After injection of an albumin-sample and a known amount of enzyme-labeled albumin, both are separated from the sample matrix by antibody-antigen-interaction. After injection of a substrate, the change in heat is a measure of analyte concentration. The less heat produced means that more albumin has been bound. An elution step regenerates the ELISA. Due to its thermal detection principle, the procedure is called TELISA (thermometric enzyme-linked immunosorbent assay). Figure 3 shows the principle of the TELISA procedure in its sandwich configuration. 

ADP AFP ab as ALAT AP ASAT ATP BQ BSA CEH CK CME COD con A CV d D E E EC ECME EDTA EIA /e FAD FET FIA G GOD G6P-DH HBg HCG adenosine diphosphate a-fetoprotein antibody antigen alanine aminotranferase alkaline phosphatase aspartate aminotransferase adenosine triphosphate benzoquinone bovine serum albumin cholesterol ester hydrolase creatine kinase chemically modified electrode cholesterol oxidase concanavalin A coefficient of variation (relative standard deviation) layer thickness diffusion coefficient enzyme potential Enzyme Classification enzyme-chemically modified electrode ethylene diamine tetraacetic acid enzyme immunoassay enzyme loading factor flavin adenine dinucleotide field effect transistor flow injection analysis amplification factor glucose oxidase glucose-6-phosphate dehydrogenase hepatitis B surface antigen human chorionic gonadotropin... 

Splenic T-cells were cultured in vitro with antibody T-cell receptor T cells from JP-8-exposed mice had reduced proliferative response T-cell-dependent antibody responses to KLH antigen injected in Freund s adjuvant were not altered by exposure to JP-8... 

Fig. 2.4 Monoclonal antibodies. After injecting the antigen and generating several clones of antibodies, the spleen containing B-cells is removed. Hybridoma cells are made by fusing spleen B-ceUs with a myeloma cell culture line. To isolate the individual hybridoma cells producing one clone of antibody, the mixed hybridoma culture is highly diluted and plated in 96-well plates with one cell or less per well...

Loiselem showed that a,a-trehalose is a true antigen. Injection of animals with a,a-trehalose produces antibodies which are incomplete and devoid of precipitating power, but, none-the-less, very specific. Sera produced in this way are able to distinguish between sucrose and a,a-trehalose a viscosity test is used. 

Polyclonal antibodies are the serum product of an immunized animal containing many different antibodies against the various mixtures of antigens injected. The antiserum is the product of many responding clones of cells and is usually heterogeneous at all levels. These levels include the specificity of the antibodies, classes and subclasses, titer, and affinity. The response to individual epitopes may be clonally diverse, and antibodies of different affinities may compete for the same epitope. This variation means that polyclonal antisera cannot be reproduced see Fig. 3). 

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