CA1132044A - OPSONISING SURFACE-BINDING .alpha..SUB.2-GLYCOPROTEIN BY DIRECT AGGLUTINATION - Google Patents
OPSONISING SURFACE-BINDING .alpha..SUB.2-GLYCOPROTEIN BY DIRECT AGGLUTINATIONInfo
- Publication number
- CA1132044A CA1132044A CA340,404A CA340404A CA1132044A CA 1132044 A CA1132044 A CA 1132044A CA 340404 A CA340404 A CA 340404A CA 1132044 A CA1132044 A CA 1132044A
- Authority
- CA
- Canada
- Prior art keywords
- glycoprotein
- opsonising
- binding
- determination
- process according
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/557—Immunoassay; Biospecific binding assay; Materials therefor using kinetic measurement, i.e. time rate of progress of an antigen-antibody interaction
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Peptides Or Proteins (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Analysing Materials By The Use Of Radiation (AREA)
Abstract
ABSTRACT
The present invention provides a process for the determination of an opsonising, surface-binding .alpha.2-glycoprotein, wherein a buffered solution of antibodies against the opsonising, surface-binding .alpha.2-glycoprotein is mixed with the sample to be tested and the turbidity formed is determined at definite intervals of time turbidimetrically or nephelometrically.
The present invention provides a process for the determination of an opsonising, surface-binding .alpha.2-glycoprotein, wherein a buffered solution of antibodies against the opsonising, surface-binding .alpha.2-glycoprotein is mixed with the sample to be tested and the turbidity formed is determined at definite intervals of time turbidimetrically or nephelometrically.
Description
~3ZO~
'I
The present invention is concerned wlth a process for the determination of an opsonising, surface-binding ~2-glycoprotein, which is hereinafter referred to as a~-SB-glycoprotein, and especially for the determination thereof in body fluids.
a2-SB-Glycoprotein is a substance which is found in blood and organ stroma. The serum protein has the - electrophoretic motility of an ~2-globulin and a sediment-ation coefficient of 12 to 14 S. Its molecular weight is about ~00,000 to 440,000. It consists of t~o subunits, which are joined together by disulphide bridges. ~he concentration thereof in the plasma of healthy persons is 300 to ~00 ~g./ml.
a2-SB-Glycoprotein is particularLy characterised by speeial adhesion properties and, like an adhesive, is able to bind other substances, for example cell debris.
a2-SB-Glycoprotein is a substrate of the acti-~ated coagulation factor ~III, i.e. of factor ~IIIa (plasma transglutaminase, fibrinoligase). It binds fibrinogen and fibrin, especially in the cold. Since, during the coagulation, it can react with fibrin, serum usually contains smaller concentrations than plasma.
A physiological property of ~2-SB-glycoprotein is its opsonising action. For example, the uptake of particulate material is promoted by the reticuloendo-thelial system (RES). Therefore, a direct correlation is assumed between the ~2-SB-glycoprotein level in the ..
.~k .~ .
, ~' ' ; ' ~3;2Q~
~! -3-plasma and the activity of the RES.
A decrease of the a2-SB-glycoprotein level occurs in certain diseases. Increased values have been observed in diseases of the connective tissue system and in metastasing tumours in advanced stages.
It is assumecl that the removal of damaged auto-logous tissue and of circulating colloidal components, for example soluble fibrin, by non-immunoLogical opson-ation is an important physiological process. For this reason, a considerable decrease of a2-SB-glycoprotein eould lead to organ failure in severely diseased patients. Therefore, the determination of a2-SB-glyco-protein is of diagnostic interest~ -Processes are kno~n for the qualitative and quantitative determination of this protein. Saba et al.
~J. Reticuloendothel~ Soc., 3, 398/1966) described a radiological ~hagocytosis test for the semi~uantitative determination of an opsonising serum factor which stim-ulates the phagocytosis of particulate material in the Kupffer cells of the liver. In this test system, the uptake of radioactiv-labelled, gelatine-coated lipid micelles in liver sections is measured. ~n immunological detection process is described hy F.A. Blumenstock et al.
(J. Reticuloendothel. Soc., 23, 119/1978). In this ease, the a2-SB-glycoprOtein is determined by means of an electro-immune assay (roeket electrophoresis). In ~Ioppe-Seylers Z. Physiol. Chemie, 359, 247/lg78, there .
is described a qu~ntit~tive method for the determin-ation of CIG ~corresponds to ~2-SB-glycoprotein), utilising the a,finity of CIG for 125-I-marked collagen, the complex formed being precipitated out with anti-CIG
antiserum. In Int. J. Cancer, 20, 1/1977, there is described an enzyme immuno assay in which plastic test tubes are coated with gelatine or specific antibodies against fibronectin and, after binding fibronectin (corresponds to a2-SB-glycoprotein) on the walls Oc the test tubes, there follows an incubation with enzyme-labelled anti-fibronectin antibody (sandwich principle). Furthermore, from J. Biol. Chem., 2~5, 5728/1970, there is known an immune-electrophoretic method for carrying out a quantitative determination.
The known methods all suffer from the disadvantage that, on the one hand, they require a relatively high expenditure of labour and, on the other hand, the carry-ing out of the test takes about 20 to 24 hours. Further-more, several working steps are necessary.
Therefore, although it has already been possible for quite a long time (1970) to determine ~2-SB-glyco-protein ~uantitatively, it has previously not been possible to develop for clinical practice a test which is simple and ~uick to carry out. Because of the special physiological properties and the diagnostic consequences proceeding therefrom, a rapid carrying out of the test would, however, be very desirable, a quick result being - ' ' ' ' ~13Z~
..
necessary, especially in acute cases, for the purpose of clinical control and of continuous monitoring.
Therefore, it is an object of the present invention to provide a rocess for the determination of a2-S~-glycoprotein ~hich can be carried out within a few minutes and in as simple a manner as possible and thus, for the first time, to provide the pre-requisite of detecting, ln a short time, changes in the amount of this substance and to make it evaluable for the purposes of diagnosis and treatment.
Thus, according to the present invention, there is provided a process for the determination of an opson-ising, surface-binding a2-glycoprotein, wherein a buffer2d solution of antibodies against the opsonising, surface-binding a2-glycoprotein is mi~ed with the sample to be investigated and the turbidity formed is determined turbidimetrically or nephelometrically at definite intervals of time.
Surprisingly, we have found that it is possible to determine a2-SB-glycoprotein turbidimetrically by an immunological method although, having regard to the adhesive properties of this substance, it wa~s to have been expected that foreign substances would be entrained and thus the proportionality bet~een the amount of the protein to be determined and the resulting turbidity could no longer be guaranteed and, furthermore, that flocculation or agglomeration phenomena would occu-r 1~3-~:044 which would make a ~uantitative determination by turbidity measurement im~ossible. This is also expressed by the term "opsonising' which literally means "preparinq for eating". This is to e~press the idea that this protein serves, as it were, for the recognition of su~stances to be removed and, by its adhesion to the cells of the reticuloendothelial system, bring about the "eating up"
of these substances with the opsonising protein adhering thereto.
The process according to the present invention is preferably carried out at a pH value of from 5 to 9.5 and especially of from 6.5 to 8.5. Substances buffer-ing in this range can be used. Buffers are preferred with a low ion strength of up to about 100 mmolar.
However, buffer solutions with greater ion strength can also be used.
In a preferred embodiment of the present invention, a polyethylene glycol is added, the water-soluble types of polyethylene glycol being preferred. An especially preferred amount is from 1 to 5fi by weight of poly-ethylene glycol, referred to the test solution.
The process is preferably carried out at a definite temperature of from 15 to 35C. and more prefer-ably of 20 to 30C In the case of lower temperatures, precipitation takes place considerably more slowly and at higher temperatures a dissociation of the immune complex ta~ses place again.
1132~
The measurement ltself can he carried out at any desired wavelength ~hich is known for turbidimetric determinations, short wavelengths, for example 334, 360 and 405 nm., being preferr~d. A nephelometric determination can aLso be used.
Measurement is carried out at definite intervals of time, for exam~le after 1 minute and after 10 minutes, the extinction di~ference of the measured values being directly proportional to the content of a2-SB-glyco-protein.
The antibody or antiserum can be ~roduced in the usual manner by injecting the a2-SB-glyco~rotein serving as antigen intradermally or intramuscularly, with re~eated administrations, into suitable experimental animals, for exam~le rabbits, sheep or goats. ~dministration can be carried out with conventional immunological adjuvants, for example Freund's adjuvant.
a2-S~-Glycoprotein is removed from human plasma by affinity chromatography and the plasma then cross-lin]~ed with glutardialdehyde to give a gel. The gel is homogenised and, using the homogenate, the antisera obtained from the experimental animals are absorbed up to immunologica~ monospecificity (tested by immune diffusion and lmmune electrophoresis). 'rhe antibodies against the a2-SB-glycoprotein not absorbed in this manner are used in th~ process.
The antiserum thus produced is diluted wlth the ., 4~
above-descri'~ed ~uffer to the desired concen.tration, optionally mixed with polyethylene glycol and stabilis-ing agents and stored in liquid or lyophilised lorm.
.~xam~les oF sta'~ilising agents which can be used include, for e~am~le, mannitol, alkali metal azides, serum albumin and the ll~ie.
The process according to the present inventlon can be used ~ot only for carrying out a manual test but also in automatic analysers. This is e~plained in more detail, with reference to the accompanying drawings, in which:
~ig.l illustrates a standard curve l~roduced wit'n a manual test;
Fig.2 illustrates a standard curve produced on an automatic analyser (ABA-100); and Fig.3 shows the correlation ~etween the determin-ati~n o a2-SB-~lycoprotein in human plasma according to the present invention, using a mamlal test, with an immune electrophoretic method (roc]cet electrophoresis).
r~he deterrnination is carried out by comparison with a standard solution o F known a2-SB-glycoproteinconcentration.
The ~rocess according to the present invention permits a rapid and e~act determination of a2-S~-glyco-protein, which only re~uires a ew minutes and can be carried out very simply with conventional photometric devices. In the sc~me way, it can also be used for auto-matic analysers.
1~32~
g Tne following ~xample i5 given for the purpose of illust~ating the pres2nt invention:
~3~ainl?1e .
A. ?roduction of the antiserum.
a2-S3-Glycoprotein was isolated by the method descri"ed in 3iochem. 3iophys. Acta, 534, 210/197~ by affinity chromatography on gelatine, bound on to cross-linked agarose.
Rahbits, sheep and goats were i~munised according to the following scheme:
, _ . .
day ¦ ~mount mode of administration ~ adjuvant 1 500 ~g.intradermally at several Freund places 7 100 ~g.intramuscularly Freund 21 100 ~g.intramuscularly Freund 49 100 ~g.intramuscularly Freund 100 ~g.intramuscularly Freund *) L~t monthly intervals.
The follo~7ing procedure was used for purification of the antisera obtained from the experimental animals 100 ml. human plasma were freed, by affinity chromato-graphy, from a2-SB-glycoprotein and then cross-linked to give a gel using glutardialdehyde(by the process described in Immunochemistry, 6, 53/1969)~ the gel obtained then being homogenised. Antiserum obtained ~-~32049~
from the experimental animals was filtered through the gel. ~fter passage through the gel, the remaining anti-serum displayed i~munological monospecificity.
- ~0 Car~yin~ out of the test --A standard solution of x2-SB-glycoprotein was produced by dissolving definite amounts thereof in 0.15 mol/litre phosphate-buffered sodium chloride sol-ution containing 1% by weight bovine serum albumin and 0.1~ by weight sodium azide.
Before use, the antiserum was brought to the desired dilution with 0.66 ~ol/litre phosphate ouffer (pH 7.4) ~hich contained 2.5,~, by ~eight polyethylene glycol and O.lC' by weight sodium azide.
For carrying out the test, 500 ~1. of the diluted antibody were introduced into a cuvette of 1 cm. layer thickness and mixed with 10 ~l. of sample solution or standard and then, at 20 to 25C., the extinction was measured against air at T~g 334 nm in a commercially available photometer 1 minute and 10 minutes after mixing. The content of a2-SB-glycoprotein was obtained by comparison of the extinction difference with a standard curve obtained in the same manner.
'
'I
The present invention is concerned wlth a process for the determination of an opsonising, surface-binding ~2-glycoprotein, which is hereinafter referred to as a~-SB-glycoprotein, and especially for the determination thereof in body fluids.
a2-SB-Glycoprotein is a substance which is found in blood and organ stroma. The serum protein has the - electrophoretic motility of an ~2-globulin and a sediment-ation coefficient of 12 to 14 S. Its molecular weight is about ~00,000 to 440,000. It consists of t~o subunits, which are joined together by disulphide bridges. ~he concentration thereof in the plasma of healthy persons is 300 to ~00 ~g./ml.
a2-SB-Glycoprotein is particularLy characterised by speeial adhesion properties and, like an adhesive, is able to bind other substances, for example cell debris.
a2-SB-Glycoprotein is a substrate of the acti-~ated coagulation factor ~III, i.e. of factor ~IIIa (plasma transglutaminase, fibrinoligase). It binds fibrinogen and fibrin, especially in the cold. Since, during the coagulation, it can react with fibrin, serum usually contains smaller concentrations than plasma.
A physiological property of ~2-SB-glycoprotein is its opsonising action. For example, the uptake of particulate material is promoted by the reticuloendo-thelial system (RES). Therefore, a direct correlation is assumed between the ~2-SB-glycoprotein level in the ..
.~k .~ .
, ~' ' ; ' ~3;2Q~
~! -3-plasma and the activity of the RES.
A decrease of the a2-SB-glycoprotein level occurs in certain diseases. Increased values have been observed in diseases of the connective tissue system and in metastasing tumours in advanced stages.
It is assumecl that the removal of damaged auto-logous tissue and of circulating colloidal components, for example soluble fibrin, by non-immunoLogical opson-ation is an important physiological process. For this reason, a considerable decrease of a2-SB-glycoprotein eould lead to organ failure in severely diseased patients. Therefore, the determination of a2-SB-glyco-protein is of diagnostic interest~ -Processes are kno~n for the qualitative and quantitative determination of this protein. Saba et al.
~J. Reticuloendothel~ Soc., 3, 398/1966) described a radiological ~hagocytosis test for the semi~uantitative determination of an opsonising serum factor which stim-ulates the phagocytosis of particulate material in the Kupffer cells of the liver. In this test system, the uptake of radioactiv-labelled, gelatine-coated lipid micelles in liver sections is measured. ~n immunological detection process is described hy F.A. Blumenstock et al.
(J. Reticuloendothel. Soc., 23, 119/1978). In this ease, the a2-SB-glycoprOtein is determined by means of an electro-immune assay (roeket electrophoresis). In ~Ioppe-Seylers Z. Physiol. Chemie, 359, 247/lg78, there .
is described a qu~ntit~tive method for the determin-ation of CIG ~corresponds to ~2-SB-glycoprotein), utilising the a,finity of CIG for 125-I-marked collagen, the complex formed being precipitated out with anti-CIG
antiserum. In Int. J. Cancer, 20, 1/1977, there is described an enzyme immuno assay in which plastic test tubes are coated with gelatine or specific antibodies against fibronectin and, after binding fibronectin (corresponds to a2-SB-glycoprotein) on the walls Oc the test tubes, there follows an incubation with enzyme-labelled anti-fibronectin antibody (sandwich principle). Furthermore, from J. Biol. Chem., 2~5, 5728/1970, there is known an immune-electrophoretic method for carrying out a quantitative determination.
The known methods all suffer from the disadvantage that, on the one hand, they require a relatively high expenditure of labour and, on the other hand, the carry-ing out of the test takes about 20 to 24 hours. Further-more, several working steps are necessary.
Therefore, although it has already been possible for quite a long time (1970) to determine ~2-SB-glyco-protein ~uantitatively, it has previously not been possible to develop for clinical practice a test which is simple and ~uick to carry out. Because of the special physiological properties and the diagnostic consequences proceeding therefrom, a rapid carrying out of the test would, however, be very desirable, a quick result being - ' ' ' ' ~13Z~
..
necessary, especially in acute cases, for the purpose of clinical control and of continuous monitoring.
Therefore, it is an object of the present invention to provide a rocess for the determination of a2-S~-glycoprotein ~hich can be carried out within a few minutes and in as simple a manner as possible and thus, for the first time, to provide the pre-requisite of detecting, ln a short time, changes in the amount of this substance and to make it evaluable for the purposes of diagnosis and treatment.
Thus, according to the present invention, there is provided a process for the determination of an opson-ising, surface-binding a2-glycoprotein, wherein a buffer2d solution of antibodies against the opsonising, surface-binding a2-glycoprotein is mi~ed with the sample to be investigated and the turbidity formed is determined turbidimetrically or nephelometrically at definite intervals of time.
Surprisingly, we have found that it is possible to determine a2-SB-glycoprotein turbidimetrically by an immunological method although, having regard to the adhesive properties of this substance, it wa~s to have been expected that foreign substances would be entrained and thus the proportionality bet~een the amount of the protein to be determined and the resulting turbidity could no longer be guaranteed and, furthermore, that flocculation or agglomeration phenomena would occu-r 1~3-~:044 which would make a ~uantitative determination by turbidity measurement im~ossible. This is also expressed by the term "opsonising' which literally means "preparinq for eating". This is to e~press the idea that this protein serves, as it were, for the recognition of su~stances to be removed and, by its adhesion to the cells of the reticuloendothelial system, bring about the "eating up"
of these substances with the opsonising protein adhering thereto.
The process according to the present invention is preferably carried out at a pH value of from 5 to 9.5 and especially of from 6.5 to 8.5. Substances buffer-ing in this range can be used. Buffers are preferred with a low ion strength of up to about 100 mmolar.
However, buffer solutions with greater ion strength can also be used.
In a preferred embodiment of the present invention, a polyethylene glycol is added, the water-soluble types of polyethylene glycol being preferred. An especially preferred amount is from 1 to 5fi by weight of poly-ethylene glycol, referred to the test solution.
The process is preferably carried out at a definite temperature of from 15 to 35C. and more prefer-ably of 20 to 30C In the case of lower temperatures, precipitation takes place considerably more slowly and at higher temperatures a dissociation of the immune complex ta~ses place again.
1132~
The measurement ltself can he carried out at any desired wavelength ~hich is known for turbidimetric determinations, short wavelengths, for example 334, 360 and 405 nm., being preferr~d. A nephelometric determination can aLso be used.
Measurement is carried out at definite intervals of time, for exam~le after 1 minute and after 10 minutes, the extinction di~ference of the measured values being directly proportional to the content of a2-SB-glyco-protein.
The antibody or antiserum can be ~roduced in the usual manner by injecting the a2-SB-glyco~rotein serving as antigen intradermally or intramuscularly, with re~eated administrations, into suitable experimental animals, for exam~le rabbits, sheep or goats. ~dministration can be carried out with conventional immunological adjuvants, for example Freund's adjuvant.
a2-S~-Glycoprotein is removed from human plasma by affinity chromatography and the plasma then cross-lin]~ed with glutardialdehyde to give a gel. The gel is homogenised and, using the homogenate, the antisera obtained from the experimental animals are absorbed up to immunologica~ monospecificity (tested by immune diffusion and lmmune electrophoresis). 'rhe antibodies against the a2-SB-glycoprotein not absorbed in this manner are used in th~ process.
The antiserum thus produced is diluted wlth the ., 4~
above-descri'~ed ~uffer to the desired concen.tration, optionally mixed with polyethylene glycol and stabilis-ing agents and stored in liquid or lyophilised lorm.
.~xam~les oF sta'~ilising agents which can be used include, for e~am~le, mannitol, alkali metal azides, serum albumin and the ll~ie.
The process according to the present inventlon can be used ~ot only for carrying out a manual test but also in automatic analysers. This is e~plained in more detail, with reference to the accompanying drawings, in which:
~ig.l illustrates a standard curve l~roduced wit'n a manual test;
Fig.2 illustrates a standard curve produced on an automatic analyser (ABA-100); and Fig.3 shows the correlation ~etween the determin-ati~n o a2-SB-~lycoprotein in human plasma according to the present invention, using a mamlal test, with an immune electrophoretic method (roc]cet electrophoresis).
r~he deterrnination is carried out by comparison with a standard solution o F known a2-SB-glycoproteinconcentration.
The ~rocess according to the present invention permits a rapid and e~act determination of a2-S~-glyco-protein, which only re~uires a ew minutes and can be carried out very simply with conventional photometric devices. In the sc~me way, it can also be used for auto-matic analysers.
1~32~
g Tne following ~xample i5 given for the purpose of illust~ating the pres2nt invention:
~3~ainl?1e .
A. ?roduction of the antiserum.
a2-S3-Glycoprotein was isolated by the method descri"ed in 3iochem. 3iophys. Acta, 534, 210/197~ by affinity chromatography on gelatine, bound on to cross-linked agarose.
Rahbits, sheep and goats were i~munised according to the following scheme:
, _ . .
day ¦ ~mount mode of administration ~ adjuvant 1 500 ~g.intradermally at several Freund places 7 100 ~g.intramuscularly Freund 21 100 ~g.intramuscularly Freund 49 100 ~g.intramuscularly Freund 100 ~g.intramuscularly Freund *) L~t monthly intervals.
The follo~7ing procedure was used for purification of the antisera obtained from the experimental animals 100 ml. human plasma were freed, by affinity chromato-graphy, from a2-SB-glycoprotein and then cross-linked to give a gel using glutardialdehyde(by the process described in Immunochemistry, 6, 53/1969)~ the gel obtained then being homogenised. Antiserum obtained ~-~32049~
from the experimental animals was filtered through the gel. ~fter passage through the gel, the remaining anti-serum displayed i~munological monospecificity.
- ~0 Car~yin~ out of the test --A standard solution of x2-SB-glycoprotein was produced by dissolving definite amounts thereof in 0.15 mol/litre phosphate-buffered sodium chloride sol-ution containing 1% by weight bovine serum albumin and 0.1~ by weight sodium azide.
Before use, the antiserum was brought to the desired dilution with 0.66 ~ol/litre phosphate ouffer (pH 7.4) ~hich contained 2.5,~, by ~eight polyethylene glycol and O.lC' by weight sodium azide.
For carrying out the test, 500 ~1. of the diluted antibody were introduced into a cuvette of 1 cm. layer thickness and mixed with 10 ~l. of sample solution or standard and then, at 20 to 25C., the extinction was measured against air at T~g 334 nm in a commercially available photometer 1 minute and 10 minutes after mixing. The content of a2-SB-glycoprotein was obtained by comparison of the extinction difference with a standard curve obtained in the same manner.
'
Claims (9)
1. Process for the determination of an opsonising, surface-binding .alpha.2-glycoprotein, wherein a buffered solution of antibodies against the opsonising, surface-binding .alpha.2-glycoprotein is mixed with the sample to be tested and the turbidity formed is determined at definite intervals of time turbidimetrically or nephelometrically.
2. Process according to claim 1, wherein a buffer solution of pH 5 to 9.5 is used.
3. Process according to claim 1, wherein a buffer solution of pH 6.5 to 8.5 is used.
4. Process according to claim 1, 2 or 3, wherein a water-soluble polyethylene glycol is added.
5. Process according to claim 1, 2 or 3, wherein 1 to 5% by weight of polyethylene glycol is added.
6. Process according to claim 1, 2 or 3, wherein the determination is carried out at a definite temperature of from 15 to 35°C.
7. Process according to claim 1, 2 or 3, wherein the determination is carried out at a definite temperature of from 20 to 30°C.
8. Process, according to claim 1, 2 or 3, wherein a water-soluble polyethylene glycol is added and the deter-mination is carried out at a definite temperature of from 15 to 35°C.
9. Process according to claims 1, 2 or 3, wherein 1 to 5%, by weight of a water soluble polyethylene glycol is added and the determination is carried out at a definite temperature of from 20 to 30°C.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE2901327A DE2901327B1 (en) | 1979-01-15 | 1979-01-15 | Method for determining an opsonizing surface-binding alpha-glycoprotein |
| DEP2901327.2 | 1979-01-15 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1132044A true CA1132044A (en) | 1982-09-21 |
Family
ID=6060586
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA340,404A Expired CA1132044A (en) | 1979-01-15 | 1979-11-22 | OPSONISING SURFACE-BINDING .alpha..SUB.2-GLYCOPROTEIN BY DIRECT AGGLUTINATION |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0013306B1 (en) |
| JP (1) | JPS5596457A (en) |
| AT (1) | ATE737T1 (en) |
| CA (1) | CA1132044A (en) |
| DE (2) | DE2901327B1 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61294366A (en) * | 1985-06-24 | 1986-12-25 | Toubishi Yakuhin Kogyo Kk | Reagent for detecting antibody specific to sugar chain and its use |
| US5371021A (en) * | 1992-07-17 | 1994-12-06 | Beckman Instruments, Inc. | Initial rate photometric method for immunoassay |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IL48804A (en) * | 1975-01-29 | 1979-05-31 | Baxter Travenol Lab | Imminological reagent comprising a mixture of polyethyleneglycol and a nonionic surfactant |
-
1979
- 1979-01-15 DE DE2901327A patent/DE2901327B1/en not_active Withdrawn
- 1979-11-02 EP EP79104289A patent/EP0013306B1/en not_active Expired
- 1979-11-02 AT AT79104289T patent/ATE737T1/en not_active IP Right Cessation
- 1979-11-02 DE DE7979104289T patent/DE2962242D1/en not_active Expired
- 1979-11-22 CA CA340,404A patent/CA1132044A/en not_active Expired
-
1980
- 1980-01-14 JP JP222680A patent/JPS5596457A/en active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| EP0013306B1 (en) | 1982-03-03 |
| DE2901327B1 (en) | 1980-07-24 |
| JPS5596457A (en) | 1980-07-22 |
| ATE737T1 (en) | 1982-03-15 |
| DE2962242D1 (en) | 1982-04-01 |
| EP0013306A1 (en) | 1980-07-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kawasaki et al. | Autoantibodies to glutamic acid decarboxylase in patients with autoimmune thyroid disease: relation to competitive insulin autoantibodies | |
| CA1180274A (en) | Immunoassay of proteins | |
| EP0840126A2 (en) | Marker and immunological reagent for dialysis-related amyloidosis, diabetes mellitus and diabetes mellitus complications | |
| US4722893A (en) | Reagents for enzyme immunoassay for apolipoprotein B | |
| EP0124320B1 (en) | Reagent for detecting antigen | |
| EP0844484B1 (en) | Method for producing a stable troponin preparation and the use thereof as a calibrator/control in immunoassays | |
| Yamada et al. | A rapid method for measuring serum amyloid A protein by latex agglutination nephelometric immunoassay | |
| US4624927A (en) | Reagent for determination of blood coagulation factor XIII | |
| CA2067584C (en) | Agglutinant complex as blood typing reagent | |
| CA1132044A (en) | OPSONISING SURFACE-BINDING .alpha..SUB.2-GLYCOPROTEIN BY DIRECT AGGLUTINATION | |
| Lauf et al. | The M-antigen in HK and LK sheep red cell membranes | |
| Besch et al. | Measurement of insulin in human sera using a new RIA kit. 1. Insulin determination in the absence of insulin antibodies—conventional assay and micro Modification2 | |
| US5811242A (en) | Marker and reagent for diabetes mellitus and diabetes mellitus complication | |
| Trenkle et al. | Effects of chemical treatment on immunochemical properties of human growth hormone and sheep interstitial cell-stimulating hormone | |
| JPS61186855A (en) | Method of measuring combining capacity of globulin forming thyroxine | |
| EP0022669B2 (en) | Method and reagent for blood typing and method for preparing this reagent | |
| CA2088404C (en) | Method for the determination of antigens or antibodies in the presence of an immune complex | |
| Macpherson | Quantitative estimation of the γc-globulin in normal and pathological cerebrospinal fluids by an immunochemical method | |
| CA2327414C (en) | Immunoassay of human medullasin and diagnosis of multiple sclerosis using the same | |
| JPH02193071A (en) | Kit of reagent for measuring haptoglobin-hemoglobin complex and measuring method of haptoglobin-hemoglobin complex using the same | |
| Ghielmi et al. | Methodological effects on the quantitation of serum ferritin by radio-and enzymoimmunoassays | |
| EP0100395B1 (en) | Reagent for determination of human urine kallikrein | |
| EP1256802B1 (en) | Method for examination of feces occult blood | |
| Kurihara et al. | Radioimmunoassay for human pancreatic ribonuclease and measurement of serum immunoreactive pancreatic ribonuclease in patients with malignant tumors | |
| Jaschonek et al. | Platelet prostacyclin binding in smokers |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MKEX | Expiry |