Chemistry: electrical and wave energy – Processes and products – Electrostatic field or electrical discharge
Patent
1979-03-14
1982-05-11
Marantz, Sidney
Chemistry: electrical and wave energy
Processes and products
Electrostatic field or electrical discharge
23230B, 424 12, G01N 3354, B01D 5702, G01N 2726
Patent
active
043292135
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to a biological indicator system based on a two-phase system containing a first biologically active component bound to the surface of a solid phase, and a liquid-saturated immobilized phase covering said surface containing a second biologically active component, capable of reacting with an unknown third biologically active component added to the immobilized phase and permitted to diffuse or migrate electrophoretically during formation of a zone containing a precipitate. The first biologically active component is capable of reacting by absorption with said precipitate formed in the immobilized phase. The presence of the third component is visualized in a suitable manner on the surface, after removal of said immobilized phase.
Serologic testing methods have a central position within the diagnostic techniques of medicine and applied biochemistry in general. This is predominantly due to the fact that such methods afford the possibility of determining minute quantities of the specific reactants in question in complex systems. Within serology there is utilized the basic fact that antibodies (ab) can specifically react with antigens (ag).
When indicating ag-ab-reaction, there are used different secondary manifestations capable of being indicated in various ways. One common method of indicating these reactions is the precipitation technique, in which precipitates are formed under certain conditions when soluble antigens and antibodies come into contact with one another. In certain cases, for example when the antigen is in particle form, agglutination reactions can be utilized, these reactions occurring when antibodies come into contact with the antigen.
In recent years, different types of markers have been used for determining ag-ab-reactions. The markers comprise radioactive isotopes, enzymes, or fluorescent substances which are bound either to the antigen or to the antibody. Indication is then effected by means of the markers in the antigen-antibody-complex, subsequent to separating said complex from non-bound antigen or anti-bodies and accompanying markers.
A specific position is held by the biological determination methods, in which a natural biological effector mechanism is utilized to indicate the antigen-antibody reaction. Among these indication methods can be mentioned complement-binding reactions and different types of neutralizing reactions of biologically active antigenic substances by blocking, for example with virus, toxins, and the like.
Antigen-antibody reactions can be quantified in different ways, by using different indication principles. The most simple of these is the end-point titration method, in which one part of an antigen-antibody system is diluted to a limit at which the indicator system can no longer indicate the reaction.
Another method is one in which indication by means of markers is used, it being necessary to separate the antigen-antibody complex. The amount of labelled reactants in the antigen-antibody complex or in the residue of the system separated therefrom is then determined.
According to one conventional method, one of the reactants, in an antigen-antibody system is incorporated, for example, in an agar gel. The other reactant is supplied in wells disposed in the gel. After some time has lapsed, a radial diffusion gradient is formed by the supplied reactant. The diffusion gradient thus formed is indicated, and the obtained quantification is determined by measuring the area of the circular indication thus formed. It is also known to have the known reactant in the ag-ab-system added to a surface below the gel phase. After diffusion and migration the unknown reactant will react with the known reactant on the surface, where it can be visualized after removing the gel phase.
Indication and quantification of antigen, utilizing known antibodies, are carried out within the field of medical diagnostics and in the follow-up various diseases.
The quantification of an unknown antigen with the aid of known antibodies is effected by immunizing a living organism
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H. Elwing et al., Int. Archs. Allergy Appl. Immun., 51, 757-762 (1976).
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