Chemistry: analytical and immunological testing – Involving an insoluble carrier for immobilizing immunochemicals – Carrier is organic
Patent
1990-09-12
1992-12-29
Ceperley, Mary E.
Chemistry: analytical and immunological testing
Involving an insoluble carrier for immobilizing immunochemicals
Carrier is organic
436534, G01N 33546, G01N 33547, G01N 33531
Patent
active
051751127
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates, by way of novel industrial products, to submicron latex particles (i.e. latex particles with a mean diameter of less than 1 micrometer) sensitized by an immunological material, that is to say particles to which an antigen or antibody is bound by covalency or adsorption. These particles are "invisible" or "transparent" when they are exposed to electromagnetic radiation such as visible light and UV, and become "visible" due to agglutination when they are exposed to said radiation after having been brought into contact with an antibody or antigen, respectively.
The present invention further relates to the method of preparing these sensitized particles and to their use in immunoassays involving reactions of the antigen/antibody type.
PRIOR ART
Immunological techniques occupy an important position in the area of biomedical analysis. They are now employed for quantifying or assaying numerous substances, among which proteins, haptens, hormones, drugs and antibodies may be mentioned in particular.
Their principle is based on the chemical binding of an antibody (or antigen) to a particulate support. The complex of particulate support and antibody or antigen constitutes a reagent; this reagent agglutinates rapidly when brought into contact with the corresponding antigen (or antibody). This agglutination is accompanied by a substantial increase in the absorbance of the medium as a function of the particle size. The particle therefore behaves as a passive agglutination-amplifying factor, resulting in a sharp increase in the absorbance.
Of the known techniques, three groups of methods have gained recognition in immunoanalysis.
The first group comprises the methods of turbidimetry and nephelometry. These are methods which are rapid to carry out but have a low sensitivity and can be used only for high protein concentrations, especially in the case of turbidimetry, which entails protein concentrations above 100 .mu.g/ml. Nephelometry is more sensitive (with use concentrations of the order of 20 .mu.g/ml) but requires very "specialized" equipment and prior treatment of the samples, which greatly restrict the possible uses.
The second group comprises the so-called LAURELL and MANCINI methods. These are simple methods but are slow to carry out and cannot be automated. They apply to protein concentrations above 20 .mu.g/ml in the case of the MANCINI method and above 5 .mu.g/ml in the case of the LAURELL method. The analysis times are several hours and often more than 24 hours.
The third group comprises the so-called labeling methods, especially the enzyme immunological methods (EIA) and radioimmunological methods (RIA). The apparatus required to carry them out is expensive, the assays consist of several reaction steps separated by washes, they are still complicated to automate and the analysis times cannot easily be reduced to less than two hours. On the other hand, these methods permit sensitivities of less than 1 ng/ml and apply to any biological substance present at concentrations advantageously in the range from 10 ng/ml to 1 mg/ml.
This situation justifies and explains the research efforts currently being made to develop novel immunoassay methods which overcome the afore-mentioned disadvantages with a view to improving the use conditions of the techniques of the first two groups when the high sensitivity of the EIA and RIA methods is not absolutely necessary.
It is known in particular that, since 1970, authors have been applying themselves to the task of refining the method of detecting immunological reactions of the antigen/antibody type belonging to the first two groups mentioned above. Thus the use of latex microparticles coated with an antigen or antibody has been envisaged for perfecting quantitative assays by turbidimetry, nephelometry and particle counting.
In the field of turbidimetry (by measurement of the light absorbance), the use of submicron latex particles (mean diameter: 0.79 micrometer) coated with anti.beta..sub.2 -microglobulin antibodies for ass
REFERENCES:
patent: 4571382 (1986-02-01), Adachi
patent: 4600698 (1986-07-01), Toth
patent: 4829101 (1989-05-01), Kraemer et al.
Amiral Jean
Laroche Pascale
Ceperley Mary E.
Diagnostica Stago
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