WO1998058259A1 - Stabilization of polypeptides for use in immunoassay procedures - Google Patents
Stabilization of polypeptides for use in immunoassay procedures Download PDFInfo
- Publication number
- WO1998058259A1 WO1998058259A1 PCT/IL1998/000278 IL9800278W WO9858259A1 WO 1998058259 A1 WO1998058259 A1 WO 1998058259A1 IL 9800278 W IL9800278 W IL 9800278W WO 9858259 A1 WO9858259 A1 WO 9858259A1
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- WIPO (PCT)
- Prior art keywords
- buffer
- sucrose
- sample
- serum
- kit
- Prior art date
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Classifications
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- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
-
- 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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
- G01N33/56927—Chlamydia
Definitions
- the present invention relates to the stabilization of immobilized polypeptides, to the improvement of immunoassay procedures.
- Immunosorbent assays have largely replaced the use of agglutination- based assays in detection of immunologically active compounds, e.g. antigens of pathogens or antibodies against them. Due to the fact that in most infections, antibodies occur in much higher concentration than the antigen, and are easily obtained in non-invasive procedures such as sampling body fluids, most diagnostic methods determine the amount of antibody to a given antigen in order to find out whether an infection with the agent that carries the antigen is ongoing. Such assays necessitate the use of antigen to capture the antibody.
- the antigen can be prepared in three major ways: a) by modifying intact pathogenic organisms, e.g.
- hepatitis B virus One example of such a situation are the various genotypes of hepatitis B virus, some of which respond well to interferon treatment while others do not.
- Another example are the two bacterial species Chlamydia pneumoniae and Chlamydia trachomatis. Prevalence for the former is high, with about 60% of the population having been infected at one time, but most infections cured, while the prevalence for the latter is less than 10%, with the possibihty of silent infections going on that may develop into chronic infections. In both situations it is desirable to find out which organism caused a present symptom before starting treatment.
- Chlamydia are gram-negative intracellular parasitic bacteria that cause acute and chronic disease in mammalian and avian species.
- the genus Chlamydia is comprised of four species: C. trachomatis, C. pneumoniae, C. precorum and C. psittaci.
- C. trachomatis is divided into 15 serovars, which may cause different diseases such as trachoma, lymphogranuloma venereum or sexually transmitted disease.
- the three different species of Chlamydia are serologically highly cross-reactive.
- serological diagnostic assays for Chlamydia use either purified elementary bodies (microimmunofluorescence, MIF and ELISA tests), lipopolysaccharide, LPS or purified major outer membrane protein, MOMP (ELISA tests) as antigens.
- Genus-specific epitopes are present in all the above antigens, resulting in low species specificity of the available tests.
- C. pneumoniae with no clinical signs
- the prevalence of anti-Chlamydia antibodies is very high. Therefore, the differentiation between C. pneumoniae and C. trachomatis specific antibodies using conventional serological screening tests (MIF, ELISA, EIA etc.) is deficient.
- sucrose as a stabilizing agent, e.g. to preserve growth- enhancing properties of serum (WO9418310), enzyme activity (JP06113847), or drug activity of interferon-beta (EP 551535).
- sucrose has not been reported for immobilized proteins or in peptide-based immunoassays.
- body fluid comprises fluids sampled from within the body, such as blood or lymph, local secretions, such as tears, semen, urine, sweat, sputum etc., samples obtained by washing (e.g. bronchiolar lavage) or swabbing, including cervical smears, and the like.
- peptide comprises peptides obtained by chemical synthesis, or by cleavage, either by chemical means or by using proteolytic enzymes, of a larger peptide or protein.
- the invention is directed to a method for improving the stability of reagents for detection of antibodies in a sample, comprising:
- Said detergent is preferably a detergent other than Tween-20.
- the first buffer is a blocking solution in an immunoassay, said blocking solution containing about 0.05 to 4 % (w/v) gelatin, about 1 to 30 % (v/v) normal rabbit serum, 1 to 20% (w/v) sucrose in PBS; and wherein the second buffer is a serum diluent containing about 0.05 to 4% (w/v) gelatin, about 1 to 30% (v/v) normal rabbit serum, and about 0.01 to 2 % (v/v) Triton-X 100 in PBS.
- the invention is further directed to a stabilizing solution for the stabilization of a polypeptide or protein attached to a solid support, comprising about 1-20% sucrose in a buffer.
- a stabilizing solution for the stabilization of a polypeptide or protein attached to a solid support comprising about 1-20% sucrose in a buffer.
- An illustrative and non- limitative example of such a solution is a solution containing about 0.03% (w/v) gelatin, about 10% (v/v) normal rabbit serum, and about 10% (w/v) sucrose in PBS.
- the invention is also directed to a buffer solution for the dilution of an antibody-containing sample to be reacted with an immobilized polypeptide, comprising about 0.05% (w/v) of a detergent other than Tween-20.
- a suitable solution comprises about 0.03% (w/v) gelatin, about 10% (v/v) normal rabbit serum, and about 0.05% Triton-X 100, in PBS.
- kits comprising the stabilization solution and/or the dilution solution of the invention. Such kits can be useful for a variety of tests, e.g. for ELISA. Practically important pathogens to be tested by the invention are Chlamydia trachomatis, or Chlamydia pneumoniae, although the invention is of course in no way limited to the testing of any specific pathogen.
- the invention also encompasses a method for detecting an infection in a mammal, said method comprising:
- the present invention provides improvements over the prior art of immunoassay using immobilized antigen. Such assays are carried out by
- IFDTTTLNPTIASGAGDVK herein designated as Ct4A (SEQ. ID NO. 1)
- VDITTLNPTIAGCGSVA herein designated as Ct4B (SEQ. ID NO. 2)
- VFDVTTLNPTIAGAGDVK herein designated as Ct4C (SEQ. ID NO. 3)
- LDVTTNATIAGKGTW herein designated as CtVDIV (SEQ. ID NO. 6)
- the following peptides designed according to the sequence of homologous MOMP domains of the related bacteria Chlamydia pneumoniae were synthesized and used in the immunoassay:
- AFPLPTDAGVATATGTKS herein designated as CpVDIII (SEQ. ID NO.
- VKGTTVNANELPNVSLSNGK herein designated as Cp2A (SEQ. ID NO.
- ELISA plates (Maxisorb, Nunc) were coated with peptides (0.3 ⁇ g/well) dissolved in 0.05M carbonate buffer pH 9.5 for 1 h at 37°C. Alternatively, coating was done overnight at room temperature or at 4°C. The peptide solutions were removed, the wells filled with blocking solution (0.03% gelatin, 10% normal rabbit serum (NRS) in 0.15 M NaCl, 0.05 M NaP0 4 , pH 7.5 [PBS]). After incubation for lh at 37°C the plates were either used immediately or after drying and incubation for various times at 37 °C as indicated in Table 1.
- blocking solution 0.03% gelatin, 10% normal rabbit serum (NRS) in 0.15 M NaCl, 0.05 M NaP0 4 , pH 7.5 [PBS]
- Sera were diluted in 1/21 in diluent (0.03% gelatin, 10% NRS, 0.05% Triton-X 100), added to the coated wells and incubated for 1 h at 37°C. After rinsing six times, with PBS/0.05% Tween-20 (PBS/Tween), anti-human IgG-conjugated HRP (diluted 1:15,000 in blocking solution) was added and incubated for 1 h at 37 °C. After rinsing as before, substrate was added and the reaction quantified by measuring OD 450.
- diluent 0.03% gelatin, 10% NRS, 0.05% Triton-X 100
- Table 2 shows an example of such a stabihty test.
- incubation of the dried coated plates at 37°C led to a drastic reduction in antibody binding (see e.g. Ct2A, Ct4A).
- other peptides like Ct4B, CplA, or Cp2A, were relatively stable even after 6 days incubation at 37 °C.
- peptide Ct4C was found to be highly unstable when Ct4C-coated and dried plates were incubated at 37 °C. Since this peptide contains a cysteine residue at its N-terminal end, and cysteine is known to be sensitive to e.g. oxidizing conditions, a modified version of peptide Ct4C lacking the cysteine residue was tested for stability and for binding to different sera. Although the modified peptide had much improved stabihty, it reacted much weaker than the original Ct4C peptide (55% less signal in immunoassay) with certain sera. It was therefore desirable to include the cysteine-containing peptide Ct4C in the immunoassay in order to maximize sensitivity.
- sucrose was added and Tween-20 omitted from the blocking solution so that the composition was 0.03% gelatin, 10% NRS, 20% sucrose in PBS. All other reagents were as in Table 2.
- the table shows the results of ELISA tests (OD 450) using the indicated peptides for coating and the indicated sera as samples.
- Serum #11 was a negative control
- serum #92 is positive for both C. trachomatis and C. pneumoniae.
- the serum mix contained 4 sera of different patients that cover most of the serovar reactivities for C. trachomatis.
- Results obtained for C. trachomatis peptides - Tables 5-16 show the results of ELISA assays carried out with immobihzed peptides stored dry for the indicated times and at the indicated temperatures. The assay was carried out as described for Table 2, with 10% sucrose and/or 0.05% Tween-20 ("PBS-T”) added to the blocking buffer where indicated.
- PBS-T 10% sucrose and/or 0.05% Tween-20
- C. trachomatis peptides In order to determine whether the presence of sucrose in the blocking buffer only affects C. trachomatis peptides (all of the tested peptides were from the VDIV region of the MOMP protein and therefore homologous in their sequence), or also affects the stabihty of other peptides, various peptides derived from the MOMP protein sequence of C. pneumoniae were tested for stability as described above for C. trachomatis peptides.
- the above experiments show that the C. trachomatis and C. pneumoniae peptides used herein display several epitopes, that these epitopes are independently affected by storage without sucrose and by the addition of Tween into the blocking solution. They further show that storage without sucrose may lead to the exposure or formation of epitopes that bind unspecifically, giving rise to high background levels and therefore raising the likelihood to obtain a false-positive output in the assay. In very rare cases, the addition of Tween-20 may help to stabilize a specific epitope, as in the case of reactivity of Cp2A to H163 (compare Tables 28 and 27).
- the composition of the serum diluent must assure specific and stable binding of the antibody to the coated antigen, while at the same time minimizing background resulting from non-specific binding. It is therefore desirable to provide the components of the diluent which achieve a maximum signal-to-noise ratio. Another requirement of the diluent is an enhanced stabihty so it can be used as component of a kit.
- the influence of detergent on antibody binding and on the stabihty of the serum and conjugate diluents was determined as follows: three sera were tested for their IgG binding activity on plates coated with the C.t.
- CD conjugate diluent
- SD serum diluent
- Tween-20 was added as indicated when preparing the diluent; for diluent stored for two weeks at the indicated temperatures, Tween was added after storage, just prior to use to both SD and CD.
- Triton-X 100 or NP-40 were able to replace Tween-20 in freshly prepared SD.
- the activity of Triton-X 100 or NP-40 was not lost after incubation of SD for 5 days at 37°C (Table 33, 37°C). Since the use of Triton-X 100 gave shghtly better reactivities after 37°C incubation, it was chosen as optimal detergent for the formulation of stable SD.
- a further test with longer periods of storage proved Triton-X 100 to be stable for ten days at 37°C (Table 34).
- Example 3 Use of the enhanced stability of immobilized peptides and serum diluent in diagnostic kits.
- kits described herein are essentially improved Enzyme- Linked Immunosorbent Assay (ELISA) kit designed for the diagnosis of C. trachomatis infections.
- ELISA Enzyme- Linked Immunosorbent Assay
- Two basic kits are exemplified, both being manipulatable to vary various of the constituents thereof to meet the users' needs.
- One kit is intended for the determination of specific C. trachomatis IgG antibodies in human sera, this being designated herein as the "IgG kit”.
- the second kit is intended for the determination of specific C. trachomatis IgA antibodies in human sera, this being designated herein as the "IgA kit”.
- the above kits of the invention will comprise at least some of the following constituents:
- a C. trachomatis antigen-coated microtiter plate with a plate cover usually of the standard multiwell type having 96 wells per plate arranged in the form of 12 columns and 8 rows, i.e., 8 wells per column for a total of 96 wells. With such plates, there will be provided 12 removable 8-well strips coated with the C. trachomatis antigen.
- trachomatis antigen will preferably be a mixture of new peptides as set forth in the aforementioned copending patent application, incorporated herein by reference, the most preferred mixtures being those designated in said apphcation as "MIX 1" (Ct2A, Ct4A, Ct4B and Ct4C) and "MIX 2" (Ct4A, Ct4B, Ct4C, Ct4D).
- MIX 1 Ct2A, Ct4A, Ct4B and Ct4C
- MIX 2 Ct4A, Ct4B, Ct4C, Ct4D
- a concentrated wash buffer usually being a concentrated PBS- Tween buffer of the standard type well known in the art.
- a conjugate diluent usually in the form of a ready-to-use buffer solution.
- a negative control which is usually a C. trachomatis IgG or IgA negative human serum in a ready-to-use form.
- a positive control which is usually a C. trachomatis IgG or IgA positive human serum in a ready-to-use form.
- a concentrated HRP-conjugate usually in the form of horseradish peroxidase (HRP) conjugated to anti-human IgG or anti- human IgA (gamma chain specific).
- HRP horseradish peroxidase
- TMB-substrate usually in the form of 3,3 ⁇ 5,5'-te-ramethyl-benzidine (DMSO) as chromagen and urea hydrogen peroxide as substrate for peroxidase (HRP).
- TMB 3,3 ⁇ 5,5'-te-ramethyl-benzidine
- DMSO dimethylsulfoxide
- HRP peroxidase
- each strip is an antigen-coated 8-well strip, with a possibility of 12 such strips per microtiter plate when using 96-well plates).
- Washing step Fill each well with wash buffer and discard the hquid; repeat this step six times.
- TMB-Substrate 1/10 in DDW Dilute the concentrated (usually lOx concentrated) TMB-Substrate 1/10 in DDW.
- ready-to-use (RTU)-TMB substrate may be used, and the dilution step omitted.
- kits more "user friendly” 1) Making the kits more "user friendly”: a. Reducing the number of washing steps. b. Adding different colors to the serum diluent and the conjugate diluent. c. Stabilizing the kit for longer shelf life, e.g. by using the improvements as set forth in example 1.
- the wash buffer that enabled only three washing steps was the one that contained non-ionic detergent, this being the above-noted PBS- Tween buffer (see constituent no. (ii) in the above hst). Further, it was found that this PBS-Tween buffer could be readily prepared in a preferred concentration of 20x concentrated, which 20x concentrated wash buffer was stable for one month at 37° C and for 11 months at 4°C
- the foUowing was carried out: The following colors were added either to the serum diluent or to the conjugate diluent and tested: violet powder, evans blue, mocca brown powders and from the food colors: blue brilliant, yellow sunset and their combination (green color).
- the serum diluent is provided with blue color and the conjugate diluent is provided with green color, thereby providing for optimal distinction between the two diluents on a color basis, while at the same time, these colors do not interfere with the assay.
- peptide mix (0.3 ⁇ g/well) was dissolved in 0.05 M phosphate buffer, pipetted into the wells and left to adsorb to the plastic for 1 hr at 37°C The unbound peptide solution was then removed and the wells filled with improved blocking solution as described in example 1. After blocking for 1 h at 37°C, the blocking solution was removed and the plates dried overnight at room temperature.
- a serum diluent composition as described in Example 1 was used (PBS containing 0.03% gelatin, 10% normal rabbit serum, 0.05% Triton-X 100).
- the IgG and IgA kits are able to detect both IgG and IgA levels in sera from C. trachomatis-infected individuals as determined by MIF MRL.
- the sensitivity and specificity of the peptide assay are high and were 94% and 90%, respectively, for IgG and 95% and 90%, for IgA.
- Results The results are summarized in Figure 2, wherein the left-hand, brick-patterned bar represents the number of sera tested positive by culture assay, the cross-hatched bars represent the number of sera tested positive by the C. trachomatis peptide assay, and the right-hand bar represents the number of sera tested negative by the C. trachomatis peptide assay for IgG antibodies.
- the open part of the right-hand bar represents the part of the sera that was tested negative by MIF assay, while the cross-hatched part of this bar represents the remaining sera tested negative by C trachomatis peptide assay for IgG antibodies.
- N represents the number of individual sera.
- Example 8 The Specificity of the IgG kit as Compared to Different MIF Tests
- the specificity of the IgG kit was determined, as compared to different MIF tests (MIFl, MIF2 and SeroFIA tests). AU the MIF-tested sera were C. trachomatis negative (C.t-) and a portion of the sera were also C. pneumoniae positive sera (Ct/C.p+). MIFl and MIF2 are standard MIF tests as noted above, while SeroFIA is a new microimmunofluorescence test for the differential detection of C. trachomatis, C. pneumoniae and C. psittaci.
- Results The results are summarized in Figure 3, from which it is apparent that the IgG kit is highly specific, and showed at least 90% specificity (horizontaUy striped, hght bars in Figure 3), as compared to the various MIF assays (black bar, MIFl assay, obhquely double-striped bar, SeroFIA assay, of Savyon, cross-striped bar, MIF 2 assay).
- the IgG kit did not cross-react with C. pneumoniae positive sera.
- Val Asp lie Thr Thr Leu Asn Pro Thr lie Ala Gly Cys Gly Ser Val 1 5 10 15
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Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
IL13355698A IL133556A0 (en) | 1997-06-19 | 1998-06-15 | Stabilization of polypeptides for use in immunoassay procedures |
EP98925910A EP0995119A1 (en) | 1997-06-19 | 1998-06-15 | Stabilization of polypeptides for use in immunoassay procedures |
JP50407899A JP2002508843A (en) | 1997-06-19 | 1998-06-15 | Stabilizing polypeptides for use in immunoassay procedures |
AU77863/98A AU7786398A (en) | 1997-06-19 | 1998-06-15 | Stabilization of polypeptides for use in immunoassay procedures |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
IL12111697A IL121116A (en) | 1997-06-19 | 1997-06-19 | Stabilization of polypeptides for use in immunoassay procedures |
IL121116 | 1997-06-19 |
Publications (1)
Publication Number | Publication Date |
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WO1998058259A1 true WO1998058259A1 (en) | 1998-12-23 |
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ID=11070279
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/IL1998/000278 WO1998058259A1 (en) | 1997-06-19 | 1998-06-15 | Stabilization of polypeptides for use in immunoassay procedures |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP0995119A1 (en) |
JP (1) | JP2002508843A (en) |
AU (1) | AU7786398A (en) |
IL (1) | IL121116A (en) |
WO (1) | WO1998058259A1 (en) |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009055382A2 (en) * | 2007-10-21 | 2009-04-30 | Yasger, Paul | One-step immunoassays exhibiting increased sensitivity and specificity |
US8173385B2 (en) | 2005-07-20 | 2012-05-08 | Kyowa Medex Co., Ltd. | Method for stabilization of peptides in a biological sample |
US20140017712A1 (en) * | 2011-03-28 | 2014-01-16 | Mitsubishi Chemical Medience Corporation | Immunological measuring method and measuring kit for whole blood sample |
WO2014100853A1 (en) | 2012-12-28 | 2014-07-03 | Cellestis Limited | A cell mediated immune response assay |
US10024937B2 (en) | 2011-09-27 | 2018-07-17 | Koninklijke Philips N.V. | Gradient amplifier with compensation for dead time and forward voltage |
WO2019060607A1 (en) * | 2017-09-20 | 2019-03-28 | Quanterix Corporation | Methods and systems for the detection of analyte molecules |
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EP0140489A1 (en) * | 1983-08-05 | 1985-05-08 | Wako Pure Chemical Industries, Ltd. | Method for stabilizing immunologically active substances immobilized on an insoluble carrier and their use in the preparation of reagents for measuring physiologically active substances |
FR2634394A1 (en) * | 1988-07-25 | 1990-01-26 | Syntex Inc | COMPOUND COMPOUND WITH DETERGENT PROPERTIES, COMPOSITIONS AND KITS CONTAINING SAME AND METHODS USING THEM TO TEMPORARILY INCREASE THE HYDROPHILIC PROPERTIES OF A MATERIAL |
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US4931385A (en) * | 1985-06-24 | 1990-06-05 | Hygeia Sciences, Incorporated | Enzyme immunoassays and immunologic reagents |
EP0374778A2 (en) * | 1988-12-19 | 1990-06-27 | Roche Diagnostics GmbH | Method for immobilising a protein in a solid phase, the protein-bearing solid phase and its use |
WO1992021769A1 (en) * | 1991-05-30 | 1992-12-10 | Abbott Laboratories | Reagents containing a nonspecific binding blocker in ion-capture binding assays |
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EP0805353A2 (en) * | 1996-05-02 | 1997-11-05 | Tosoh Corporation | Immunoreaction agent and process for production thereof |
-
1997
- 1997-06-19 IL IL12111697A patent/IL121116A/en not_active IP Right Cessation
-
1998
- 1998-06-15 JP JP50407899A patent/JP2002508843A/en active Pending
- 1998-06-15 EP EP98925910A patent/EP0995119A1/en not_active Withdrawn
- 1998-06-15 WO PCT/IL1998/000278 patent/WO1998058259A1/en not_active Application Discontinuation
- 1998-06-15 AU AU77863/98A patent/AU7786398A/en not_active Abandoned
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EP0140489A1 (en) * | 1983-08-05 | 1985-05-08 | Wako Pure Chemical Industries, Ltd. | Method for stabilizing immunologically active substances immobilized on an insoluble carrier and their use in the preparation of reagents for measuring physiologically active substances |
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US5187066A (en) * | 1990-02-14 | 1993-02-16 | Syntex (U.S.A.) Inc. | Methods for detecting amphiphilic antigens |
WO1992021769A1 (en) * | 1991-05-30 | 1992-12-10 | Abbott Laboratories | Reagents containing a nonspecific binding blocker in ion-capture binding assays |
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EP0805353A2 (en) * | 1996-05-02 | 1997-11-05 | Tosoh Corporation | Immunoreaction agent and process for production thereof |
Cited By (15)
Publication number | Priority date | Publication date | Assignee | Title |
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US8173385B2 (en) | 2005-07-20 | 2012-05-08 | Kyowa Medex Co., Ltd. | Method for stabilization of peptides in a biological sample |
WO2009055382A2 (en) * | 2007-10-21 | 2009-04-30 | Yasger, Paul | One-step immunoassays exhibiting increased sensitivity and specificity |
WO2009055382A3 (en) * | 2007-10-21 | 2009-09-24 | Yasger, Paul | One-step immunoassays exhibiting increased sensitivity and specificity |
US10191038B2 (en) * | 2011-03-28 | 2019-01-29 | Lsi Medience Corporation | Immunological measuring method and measuring kit for whole blood sample |
US20140017712A1 (en) * | 2011-03-28 | 2014-01-16 | Mitsubishi Chemical Medience Corporation | Immunological measuring method and measuring kit for whole blood sample |
US10024937B2 (en) | 2011-09-27 | 2018-07-17 | Koninklijke Philips N.V. | Gradient amplifier with compensation for dead time and forward voltage |
WO2014100853A1 (en) | 2012-12-28 | 2014-07-03 | Cellestis Limited | A cell mediated immune response assay |
EP2939025A1 (en) | 2012-12-28 | 2015-11-04 | Cellestis Limited | A cell mediated immune response assay |
EP2939025A4 (en) * | 2012-12-28 | 2016-04-20 | Cellestis Ltd | A cell mediated immune response assay |
EP3421997A1 (en) * | 2012-12-28 | 2019-01-02 | Cellestis Limited | A cell mediated immune response assay |
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CN110118871A (en) * | 2012-12-28 | 2019-08-13 | 塞尔雷斯蒂斯有限公司 | Cell-mediated immune response test |
US10564150B2 (en) | 2012-12-28 | 2020-02-18 | Cellestis Limited | Cell mediated immune response assay |
EP3745128A1 (en) * | 2012-12-28 | 2020-12-02 | Cellestis Limited | A cell mediated immune response assay |
WO2019060607A1 (en) * | 2017-09-20 | 2019-03-28 | Quanterix Corporation | Methods and systems for the detection of analyte molecules |
Also Published As
Publication number | Publication date |
---|---|
IL121116A (en) | 2000-12-06 |
JP2002508843A (en) | 2002-03-19 |
AU7786398A (en) | 1999-01-04 |
EP0995119A1 (en) | 2000-04-26 |
IL121116A0 (en) | 1997-11-20 |
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