Chemistry: analytical and immunological testing – Biospecific ligand binding assay – Utilizing isolate of tissue or organ as binding agent
Patent
1984-05-30
1987-05-26
Nucker, Christine M.
Chemistry: analytical and immunological testing
Biospecific ligand binding assay
Utilizing isolate of tissue or organ as binding agent
436518, 436520, 436521, 436528, G01N 33567, G01N 33543, G01N 33555, G01N 33556
Patent
active
046686371
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to the field of biology and more particularly to the detection, by erythroadsorption, of a biological substance immobilised on a support.
It relates in particular to improvements to the process for detection and determination of a biological substance by erythroadsorption described in patent application FR No. 80/15,293.
In this patent application FR No. 80/15,293 a process for detecting and determining a biological substance by erythroadsorption has already been described. This process employs, as a reagent for determination, the product of coupling a specific ligand with a ligand capable of reacting with erythrocytes, and erythrocytes as a developer.
This process for detection and determination of a biological substance by erythroadsorption consists in:
(1) immobilising on a support a substance having a fixing affinity for the biological substance to be determined;
(2) incubating this substance with the liquid medium containing the biological substance to be determined;
(3) incubating, after washing, the resulting reaction medium with the product of coupling of a specific igand with a ligand capable of reacting with erythrocytes.
(4) Adding erythrocytes; and
(5) Determining the erythrocyte adsorption.
This process is suitable for determining, as biological substances, antigens, antibodies, haptens, hormones immunoglobulins and other substances of biological interest.
According to the teaching of this Patent FR No.80 15,293, the determination of erythroadsorption can be carried out in several ways.
For example, it is possible to establish visually that the red cells are adsorbed at the surface of the support on which the substance having a fixing affinity for the biological substance to be determined has been immobilised. In this case, the process makes it possible to identify a particular biological substance in a given biological liquid. If, on the other hand, the biological liquid does not contain the particular biological substance, the erythrocytes are not adsorbed and form a residue at the bottom of the receptacle, for example wells of the microplate. The process for determination by erythroadsorption according to this Patent FR No. 80/15,293 also makes it possible to determine a given biological substance quantitatively. For this purpose, the red cells which have not reacted are removed, for example by suction with a pipette. The adsorbed erythrocytes are then lysed, for example with distilled water, and the substances liberated by the red cells, for example haemoglobin or the substances introduced artificially by the experimenter, are then determined by spectrophotometry.
Haemoglobin can also be determined by an enzymatic reaction. It is possible, for example, to employ one of the peroxidase substrates, such as ortho-dianisidine, or ortho-phenylenediamine. The reading is also carried out by spectrophotometry, at 400 nm for orthodianisidine and 492 nm for ortho-phenylenediamine.
The quantity of substances liberated by the red cells, for example the quantity of liberated haemoglobin, is proportional to the quantity of the substance to be determined, which makes it possible to obtain, for example, the estimate of an antigen or an antibody present in the sample by referring to a standard range of haemolysis of red cells produced under the same conditions.
The quantitative determination of erythroadsorption as defined above therefore requires the removal of red cells which have not reacted and the determination of the substances liberated by the red cells or substances introduced artificially by the experimenter.
In this process, the biological substance to be determined is immobilised on a support by specific fixation, that is to say through the intermediacy of a substance having a fixing affinity for the biological substance to be determined.
It is also possible to determine by erythroadsorption, using the same operating procedure, substances which are fixed on a support by any other means, for example by passive adsorption or by chemical bonding.
It has now been found t
REFERENCES:
patent: 3708572 (1973-01-01), Pegtom et al.
patent: 3987159 (1976-10-01), Spona et al.
patent: 4371515 (1983-02-01), Chu
patent: 4493793 (1985-01-01), Chu
patent: 4526871 (1985-07-01), Avrameas et al.
Pereira, J. Immunol. Methods, 63(1983) 25-34.
Lawny et al., Chemical Abstracts, 90(1979) #70358e.
Avrameas Stratis
Guesdon Jean-Luc
Institut Pasteur
Nucker Christine M.
LandOfFree
Method for detecting and dosing by erythroadsorption a biologica does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Method for detecting and dosing by erythroadsorption a biologica, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Method for detecting and dosing by erythroadsorption a biologica will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-703721