CN106885908B - The detection kit and its detection method of blood-serum P SMD4 albumen and application - Google Patents
The detection kit and its detection method of blood-serum P SMD4 albumen and application Download PDFInfo
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- CN106885908B CN106885908B CN201510979741.3A CN201510979741A CN106885908B CN 106885908 B CN106885908 B CN 106885908B CN 201510979741 A CN201510979741 A CN 201510979741A CN 106885908 B CN106885908 B CN 106885908B
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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
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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
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
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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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
Abstract
The invention belongs to immunologys and field of biotechnology, detection kit and its detection method and application more particularly to a kind of blood-serum P SMD4 albumen, the ELISA detection kit and its detection method of PSMD4 albumen provided by the invention can be used for serodiagnosis and the tumor screening of Several Kinds of Malignancy (especially lung cancer, gastric cancer and colorectal cancer).Operation of the present invention is simple and convenient, can detect the content of the PSMD4 albumen in malignant neoplastic disease human serum accurately, in high sensitivity, provides a kind of new means and method for clinical examination and basic research.
Description
Technical field
The invention belongs to immunologys and field of biotechnology, and in particular to a kind of ELISA detection kit of PSMD4 albumen
And its detection method and answering in Several Kinds of Malignancy (lung cancer, gastric cancer and colorectal cancer) serodiagnosis and tumor screening
With.
Background technique
Cancer (cancer) refers to all malignant tumours.According to the statistics of the World Health Organization, global cancer patient is new every year
Increase more than 1,400 ten thousand, China becomes the country for being affected by cancer most serious in the world.The cancer morbidity of China's Mainland almost accounts for
The half in the whole world.Cancer has also come second in the various causes of the death in Chinese range, has then ranked the first in city.
Wherein lung cancer case fatality rate rises most fast, has come first in mortality of malignant tumors.Followed by liver cancer, gastric cancer and knot are straight
Intestinal cancer.It is a concern that coming 10 years, Chinese cancer morbidity will continue to rise with the death rate.Therefore, the morning of cancer
Phase diagnosis is of great significance to the life span and reduction mortality that extend patient.
The diagnosis of cancer at present relies primarily on laboratory inspection, imageological examination and pathologic finding and combines.But shadow
Picture checks and pathologic finding is all had no idea, and Large-scale Screening goes out earlier stage cancer patients.And laboratory checks and then needs by more
Kind tumour immunity marker detection carrys out guiding clinical diagnosis.Currently used for there are many kinds of clinical tumor markers, if AFP is liver
Cancer most has the index of diagnostic value, the S-CEA (CEA) of colon cancer；The gastric juice sulfoglucoprotein (FSA) of gastric cancer, gastric cancer phase
Close antigen (GCAA), the prostate-specific antigen (PSA) of prostate cancer etc..But according to research in recent years, various tumour marks
All there is be easy to fail to pinpoint a disease in diagnosis and the problem of mistaken diagnosis will object.The clinical tumor markers that specificity is good not yet, sensibility is high at present
It can be used for the Large-scale Screening of kinds cancer patient.It is therefore, clinical that there is still a need for highly sensitive malignant tumour specific markers
The preliminary screening of large-scale crowd for cancer patient, and then improve the early diagnostic rate of malignant tumour.
The research of malignant tumour blood serum designated object is always the emphasis of scientists study, but most of research achievements are difficult to
Meet the needs of practical application.The serological index for exploring hypersensitivity and specificity is particularly important.
Research object-PSMD4 albumen in the present invention, be a proteasome 26S non ATP enzyme subunit 4 (GeneID:
5710).The molecule contain two sections of 15 amino acid ubiquitin interaction block (ubiquitin interactingmotif,
UIM), can selective binding ubiquitin protein, mediating protein degradation.Existing research report, PSMD4 unconventionality expression and inflammation
Property the diseases such as enteropathy, ulcerative colitis it is related, expression and biological function Shang Bushi of the PSMD4 in malignant tumour are distinguished
Chu.Have studies have shown that PSMD4 may it is related with cell differentiation and body development (referring to document: Hamazaki, J., et al.,
Rpn10-mediated degradation of ubiquitinated proteins is essential for mouse
development.Mol Cell Biol,2007.27(19):p.6629-38).We have found that PSMD4 is an important cancer
Albumen, and blood can be released into.The applicant is with regard to PSMD4 as in the new application application of prognosis in hcc diagnosis marker
State patent CN201310005491.4, it is entitled " PSMD4 albumen is preparing the application in prognosis in hcc assessment kit ",
Publication No. CN103091493A.
There is no literature reported on PSMD4 albumen is accurately detected in affinity antibody to SpA sample at present；Also it there is no for swollen
PSMD4 protein content provides a kind of accurate, easy, high sensitivity detection means in tumor patients serum's sample；Also it there is no document
Report the application of the ELISA detection kit and such kit of PSMD4 albumen in tumor serology diagnosis and screening.
Summary of the invention
The object of the present invention is to provide a kind of ELISA detection kits of PSMD4 albumen；It is another object of the present invention to mention
For the detection method using mentioned reagent box；The third object of the present invention be to provide mentioned reagent box Several Kinds of Malignancy (such as
Lung cancer, gastric cancer, colorectal cancer etc.) application in serodiagnosis and screening.
The present invention is directed to for PSMD4 protein content in clinical and laboratory Several Kinds of Malignancy blood serum sample provide it is accurate,
Simplicity, high sensitivity and the detection means that can be widely used.
The present inventor detects PSMD4 albumen in a variety of evils after extensive and in-depth study, using enzyme linked immunosorbent assay
Expression in property affinity antibody to SpA, and confirm that there are PSMD4 albumen in the serum of Several Kinds of Malignancy patient for the first time
High expression.
First aspect present invention, provides a kind of ELISA detection kit of PSMD4 albumen, and the ELISA detects examination
Agent box includes:
(1) it is coated with the ELISA ELISA Plate of PSMD4 antibody, the coated antibody is the Anti-TNF-α of rabbit-anti people PSMD4
Body；Preferably, the polyclonal antibody of rabbit-anti people PSMD4 is purchased from PTG company；Optimal, antibody working concentration is 2 μ g/ml.
(2) antibody for detecting PSMD4 antigen is the monoclonal antibody of mouse anti human PSMD4；Preferably, mouse anti human
The monoclonal antibody of PSMD4 is purchased from PTG company；Optimal, the working concentration for detecting antibody is 0.5 μ g/ml.
(3) ELIAS secondary antibody, the goat anti-mouse IgG of HRP (horseradish peroxidase) label；Preferably, the mountain of HRP label
Goat anti-mouse igg is purchased from PTG company；It is optimal, diluted concentration 1:5000.
(4) standard protein is recombined human PSMD4 fusion protein；Preferably, recombined human PSMD4 fusion protein is public purchased from PTG
Department.
(5) kit described in further includes having: conventional sample diluting liquid, coating buffer, confining liquid, ELISA enzyme mark
Plate cleaning solution, antibody diluent, developing solution and terminate liquid；
The kit, ELISA ELISA Plate are commercially available ELISA ELISA Plate, preferably Germany greiner company
ELISA ELISA Plate (the 96 removable ELISA Plate #650180 of hole single)；
It is coated with buffer, selects the 1 × PBS, pH:9.6 of sigam company；
Confining liquid selects the confining liquid containing 3% bovine serum albumin(BSA)+PBS solution；
ELISA ELISA Plate cleaning solution is preferably 1 × PBS solution containing 0.05%Tween-20；
Sample diluting liquid: being preferably 1 × PBS when sample is serum, pH:7.4；
Antibody diluent selects 1 × PBS；
Developing solution selects 3,3 ', 5,5 '-tetramethyl benzidines (TMB) of sigam company；
Terminate liquid is preferably the sulfuric acid solution of 2mol/L.
A kind of ELISA detection kit of PSMD4 albumen of the present invention, reagent used, condition advanced optimize:
(1) optimization of coated antibody: kit of the invention selects different antibody to be coated with respectively: rabbit-anti people PSMD4's
Polyclonal antibody (PTG company, the # of polyclonal antibody (Abnova company, #H00005710-AP11-1), rabbit-anti people PSMD4
14899-1-AP), the monoclonal antibody (Abnova company, #H00005710-AP11-2) of the anti-human PSMD4 of mouse, the anti-human PSMD4 of mouse
Monoclonal antibody (PTG company, #66179-1-Ig), detectable concentration select 7.5 μ g/ml, 5 μ g/ml, 2.0 μ g/ml, 1 μ g/
Ml, 0.5 μ g/ml, 0.25 μ g/ml.As a result, it has been found that selecting polyclonal antibody (Abnova company, the # of rabbit-anti people PSMD4
H00005710-AP11-1), monoclonal antibody (Abnova company, the #H00005710-AP11-2 of the anti-human PSMD4 of mouse；PTG is public
Department, #66179-1-Ig) it is used as coated antibody to there is apparent non-specific responding, the concentration and absorption values of standard items do not have
There is correlation；But use the polyclonal antibody (PTG company, #14899-1-AP) of rabbit-anti people PSMD4 then without apparent non-spy
Opposite sex reaction, the concentration and absorbance of standard items have preferable correlation.By optimizing repeatedly, 2 μ g/ml of antibody working concentration
To be best, the concentration of standard items and the correlation of absorption values are best.
(2) optimization of antibody: kit of the invention is detected, selects different antibody as detection antibody respectively: working as packet
Two kinds of polyclonal antibody (Abnova companies for being rabbit-anti people PSMD4 by antibody；#H00005710-AP11-1；PTG company, #
14899-1-AP) two kinds of monoclonal antibodies (Abnova company, #H00005710-AP11-2 of the anti-human PSMD4 of Shi Xuanyong mouse；PTG
Company, #66179-1-Ig) respectively as detection antibody；When two kinds of monoclonal antibodies that coated antibody is the anti-human PSMD4 of mouse
(Abnova company, #H00005710-AP11-2；PTG company, #66179-1-Ig) more than two kinds grams of Shi Xuanyong rabbit-anti people PSMD4
Grand antibody (Abnova；#H00005710-AP11-1；PTG company, #14899-1-AP).Detectable concentration selects 2.0 μ g/ml, 1 μ
G/ml, 0.5 μ g/ml, 0.25 μ g/ml, 0.1 μ g/ml.As a result, it has been found that the polyclonal antibody of coated antibody rabbit-anti people PSMD4
(Abnova company；#H00005710-AP11-1) with two kinds of monoclonal antibodies (Abnova company, # of the anti-human PSMD4 of mouse
H00005710-AP11-2；PTG company, #66179-1-Ig) compatibility when apparent non-specific responding, the concentration of standard items is presented
There is no correlation with absorption values；The polyclonal antibody (PTG company, #14899-1-AP) of rabbit-anti people PSMD4 is anti-human with mouse
Obvious non-specific responding is equally existed when monoclonal antibody (Abnova company, #H00005710-AP11-2) compatibility of PSMD4,
The concentration and absorption values of standard items do not have correlation；Respectively by the monoclonal antibody of two kinds of anti-human PSMD4 of coated antibody mouse
(Abnova company, #H00005710-AP11-2；PTG company, #66179-1-Ig) with two kinds of Anti-TNF-αs of rabbit-anti people PSMD4
Body (Abnova；#H00005710-AP11-1；PTG company, #14899-1-AP) compatibility, as a result there is also obvious non-specific anti-
It answers, the concentration and absorption values of standard items do not have correlation.It is verified and is found by multiple permutation and combination, only selecting rabbit-anti people
The polyclonal antibody (PTG company, #14899-1-AP) and mouse anti human PSMD4 monoclonal antibody (PTG company, #66179-1- of PSMD4
Ig) when compatibility, the concentration and absorbance of standard items have preferable correlation.Subsequent process optimizes repeatedly, when detection antibody
When working concentration is 0.5 μ g/ml, the concentration of standard items and the correlation of absorption values are best.
(3) optimization of confining liquid: selecting different confining liquids respectively, contains 1% bovine serum albumin(BSA)+PBS solution, contains 2%
Bovine serum albumin(BSA)+PBS solution contains 3% bovine serum albumin(BSA)+PBS solution, contains 4% bovine serum albumin(BSA)+PBS solution, contains
5% bovine serum albumin(BSA)+PBS solution, off-period select room temperature 1 hour, and room temperature 2 hours, 4 DEG C overnight.As a result, it has been found that best
It is optimal combination that confining liquid and off-period, which are containing 3% bovine serum albumin(BSA)+PBS solution confining liquid and room temperature closing 2 hours,.
(4) optimization of ELIAS secondary antibody: according to the different ELIAS secondary antibodies for selecting different genera of detection antibody, detection antibody is
Mouse anti human PSMD4 monoclonal antibody (Abnova company, #H00005710-AP11-2；PTG company, #66179-1-Ig) Shi Xuanyong goat
The IgG antibody (Jackson company #715-005-150) of anti-mouse HRP label and the IgG antibody of goat anti-mouse HRP label
(PTG company #SA00001-1)；Detecting antibody is the mostly anti-(Abnova of rabbit-anti people PSMD4；#H00005710-AP11-1；PTG is public
Department, #14899-1-AP) Shi Xuanyong goat antirabbit HRP label IgG antibody (PTG company #SA00001-15).Antibody dilution times
Number selection 1:1000,1:2000,1:3000,1:5000,1:10000.The IgG that goat anti-mouse HRP is marked in combined authentication
The IgG antibody (PTG company #SA00001-15) of antibody (Jackson company #715-005-150) and goat antirabbit HRP label
Two groups of results do not show preferable correlation, and there are non-specific respondings.So the medium and small anti-human PSMD4 monoclonal antibody of mouse is combined in verifying
(PTG company, #66179-1-Ig) is as the IgG antibody compatibility that detection antibody and ELIAS secondary antibody are that goat anti-mouse HRP is marked
Preferable correlation (PTG company #SA00001-1) as the result is shown, the diluted concentration of 1:5000 is best.
(5) optimization of sample diluting liquid: different sample diluting liquids, sample diluting liquid when serum sample detects are selected respectively
It selects 1 × PBS (pH:7.4) or contains 0.1%BSA, 1 × PBS solution of 0.5%Tween-20.As a result, it has been found that best serum sample
Dilution is that 1 × PBS (pH:7.4) is better than containing 0.1%BSA, 1 × TBS solution of 0.5%Tween-20.
Second aspect of the present invention provides the detection method of the ELISA detection kit using above-mentioned PSMD4 albumen.
Before detection, detection ELISA ELISA Plate is prepared first: being coated with buffer for the antibody (Anti-TNF-α of rabbit-anti people PSMD4
Body) it is diluted to 2.0ug/ml, 100 μ l are added in every hole of ELISA ELISA Plate, sealing plate is placed on 4 DEG C of overnight incubations, spare.
Of the invention detection method includes the following steps: serum sample is diluted with sample diluting liquid with 1:1, with 100 holes μ l/
Volume sample-adding, be incubated at room temperature 2 hours；PSMD4 recombinant protein is diluted to various concentration gradient with sample diluting liquid, with 100 μ l/
The volume in hole is loaded, and is incubated at room temperature 2 hours；It gets rid of the liquid in each hole, is added ELISA ELISA Plate cleaning solution, every 300 μ l of hole,
Washing 3-5 times, drying；PSMD4 detection antibody is diluted to 0.5 μ g/ml, every 100 μ l of hole, is incubated at room temperature 1 hour；Get rid of each hole
In liquid, be added ELISA ELISA Plate cleaning solution, every 300 μ l of hole, wash 3-5 time, dry；The goat of addition HRP label resists small
Mouse IgG, every 100 μ l of hole are incubated at room temperature 40 minutes；The liquid in each hole is got rid of, ELISA ELISA Plate cleaning solution, every hole 300 is added
μ l is washed 6-8 times, drying；TMB, every 100 μ l of hole is added, room temperature is protected from light incubation 10-20 minutes, and 2mol/L sulfuric acid solution is added
Terminate reaction, every 50 μ l of hole；(450nm) measures OD value in microplate reader.
PSMD4 recombinant protein is diluted to various concentration gradient with sample diluting liquid, can need sets itself according to detection
Concentration gradient, in a preferred embodiment of the invention, concentration gradient are as follows: 10ng/ml, 3ng/ml, 1ng/ml, 0.3ng/
Ml, 0.1ng/ml, 0.03ng/ml, 0.01ng/ml.
Third aspect present invention provides the ELISA detection kit of above-mentioned PSMD4 albumen in Several Kinds of Malignancy blood
The clear application learned in diagnosis.
The present invention provides a kind of ELISA detection kit of PSMD4 albumen preparation malignant tumour serodiagnosis or
Application in kit for screening.
The detection kit of blood-serum P SMD4 albumen of the invention can simultaneously specifically, accurately, it is qualitative and quantitatively
Detection, diagnosis, screening Several Kinds of Malignancy.
The malignant tumour is in a preferred embodiment of the invention lung cancer, gastric cancer and/or colorectal cancer etc..
The ELISA detection kit of PSMD4 albumen of the invention is in preparation malignant tumour serodiagnosis or screening agent
Application in box, the ELISA detection kit of the PSMD4 albumen include above-mentioned kit forms.
The ELISA detection kit of PSMD4 albumen of the invention is in preparation malignant tumour serodiagnosis or screening agent
Application in box, method of the kit for serodiagnosis or screening includes above-mentioned detection method step.
Beneficial effects of the present invention:
1) accurate: the ELISA detection kit of humanized's PSMD4 albumen of commercial-free currently on the market is examined through document
The method that rope does not have quantitative detection humanized's PSMD4 protein content, previous work finds that PSMD4 is cancer protein, but at present can not
PSMD4 in Several Kinds of Malignancy patients serum is measured.This ELISA kit can accurately detect Several Kinds of Malignancy disease
The content of PSMD4 albumen in human serum eliminates the semidefinites such as immunohistochemistry, immunofluorescence as a result by microplate reader quantitative analysis
The subjectivity of amount method.
2) high sensitivity: the PSMD4 albumen detected with this method is minimum to be apparently higher than general to 10pg/ml, sensibility
The semi-quantitative methods such as logical Western blot and immunohistochemistry.
3) simple and convenient: agents useful for same and experiment consumptive material are commercially available commercially produced product in this method, are easy to get；Detection
In only need pipettor and microplate reader to be loaded and read, common laboratory and hospital can carry out this detection.
ELISA kit provided by the invention is simple to operate, can detect accurately, in high sensitivity a variety of pernicious swollen
The content of PSMD4 albumen in tumor patients serum；It is provided for screening primary malignant tumour patient and basic research a kind of new
Means and method.
Kit of the invention is mainly used for the quantitative inspection of PSMD4 albumen in the blood serum sample of Several Kinds of Malignancy patient
It surveys, can operate with the PSMD4 albumen in various biological samples (such as cell culture supernatant, cell pyrolysis liquid) in basic research
Detection.
The present invention does not need complex instrument when detecting, and is easy to promote and apply in research institutions and medical institutions, can advise greatly
Mould detects clinical samples, is quickly obtained the relevant mass data of people's PSMD4 albumen and information, is the morning of Several Kinds of Malignancy patient
Phase diagnosis and screening provide clinical reference value, have a vast market foreground, biggish economic and social benefit.
Detailed description of the invention
Fig. 1 is double-antibodies sandwich ELISA detection PSMD4 fusion protein as a result, wherein A figure is histogram, and B figure is
Scatter plot (linear related).
Fig. 2 be double-antibodies sandwich ELISA detection normal population and Serum of Patients with Lung Cancer in PSMD4 albumen as a result,
Normal (normal healthy controls)；Lung cancer (lung cancer)；
Fig. 3 be double-antibodies sandwich ELISA detection normal population and Serum Obtained From Advance Gastric Cancer in PSMD4 albumen as a result,
Normal (normal healthy controls)；Gastric cancer (gastric cancer)；
Fig. 4 is the knot of PSMD4 albumen in double-antibodies sandwich ELISA detection normal population and serum in patients with colorectal
Fruit, normal (normal healthy controls)；Colorectal cancer (colorectal cancer).
Specific embodiment
Now in conjunction with embodiment and attached drawing, the invention will be further described, but implementation of the invention is not limited to that.
The polyclonal antibody of rabbit-anti people PSMD4, Abnova company；H00005710-AP11-1
The monoclonal antibody of mouse anti human PSMD4, Abnova company；H00005710-AP11-2
The polyclonal antibody of rabbit-anti people PSMD4, PTG company；14899-1-AP
The monoclonal antibody of mouse anti human PSMD4, PTG company；66179-1-Ig
The goat anti-mouse IgG of HRP label, PTG company；SA00001-1
The goat anti-rabbit igg of HRP label, PTG company；SA00001-15
The goat anti-mouse IgG of HRP label, Jackson company；715-005-150
Recombined human PSMD4 fusion protein, PTG company；ag6691
3,3 ', the 5,5 '-tetramethyl benzidines (TMB) of developing solution selection sigam company.
Embodiment 1:
Detect the preparation of ELISA ELISA Plate: coating 1 × PBS of buffer, pH:9.6 are by antibody (more grams of rabbit-anti people PSMD4
Grand antibody) it is diluted to 2.0ug/ml, 100 μ l are added in every hole of ELISA ELISA Plate, sealing plate is placed on 4 DEG C of overnight incubations, standby
With.
The detection of serum sample and recombinant protein: getting rid of the liquid in each hole, and ELISA ELISA Plate cleaning solution is added and (contains
1 × PBS solution of 0.05%Tween-20), every 300 μ l of hole is washed 2-3 times, drying；With the PBS for containing 3% bovine serum albumin(BSA)
The confining liquid of solution is closed, and is loaded with the volume in the hole 200ul/, is closed 2 hours at room temperature: being got rid of the liquid in each hole, is added
ELISA ELISA Plate cleaning solution, every 300 μ l of hole are washed 2-3 time, are dried, and serum sample is with 1 × PBS of sample diluting liquid (pH:7.4)
It is diluted with 1:1, is loaded with the volume in 100 holes μ l/, is incubated at room temperature 2 hours；PSMD4 recombinant protein is diluted to sample diluting liquid
Various concentration gradient is loaded with the volume in 100 holes μ l/, is incubated at room temperature 2 hours；The liquid in each hole is got rid of, ELISA enzyme is added
Target cleaning solution, every 300 μ l of hole are washed 3-5 times, drying；PSMD4 detection antibody is diluted to 0.5 μ g/ml, every 100 μ l of hole,
Incubation at room temperature 1 hour；The liquid in each hole is got rid of, ELISA ELISA Plate cleaning solution is added, every 300 μ l of hole is washed 3-5 times, got rid of
It is dry；The goat anti-mouse IgG of HRP label is added, every 100 μ l of hole is incubated at room temperature 40 minutes；The liquid in each hole is got rid of, is added
ELISA ELISA Plate cleaning solution, every 300 μ l of hole are washed 6-8 times, drying；TMB, every 100 μ l of hole is added, room temperature, which is protected from light, is incubated for 10-
20 minutes, 2mol/L sulfuric acid solution is added and terminates reaction, every 50 μ l of hole；(450nm) measures OD value in microplate reader.
Embodiment 2: optimization
1, the optimization of coated antibody concentration:
Different coated antibody rabbit-anti people PSMD4 polyclonal antibodies (PTG company and Abnova company) is selected respectively, is selected
Various concentration (7.5 μ g/ml, 5 μ g/ml, 2.0 μ g/ml, 1 μ g/ml, 0.5 μ g/ml, 0.25 μ g/ml) is coated with ELISA ELISA Plate,
It is detected according to the operating procedure in embodiment 1, known various concentration PSMD4 protein standard substance is added.According to acquisition
The conduct the closest of OD value, selection blank group OD value minimum, and the linear relationship between OD value and standard protein concentration is best
Antibody (PTG company) coated antibody concentration is 2.0 μ g/ml.
2, the optimization of confining liquid:
Different confining liquids is selected respectively, contains 1% bovine serum albumin(BSA)+PBS solution, contains 2% bovine serum albumin(BSA)+PBS
Solution contains 3% bovine serum albumin(BSA)+PBS solution, contains 4% bovine serum albumin(BSA)+PBS solution, containing 5% bovine serum albumin(BSA)+
PBS solution, off-period select room temperature 1 hour, and room temperature 2 hours, 4 DEG C overnight.It is examined according to the operating procedure in embodiment 1
It surveys, known various concentration PSMD4 protein standard substance is added, select blank group OD value minimum, and OD value and standard protein concentration
Between the closest best confining liquid of conduct and off-period of linear relationship.Containing 3% bovine serum albumin(BSA)+PBS solution
Confining liquid and room temperature close 2 hours as optimal combination.
3, the optimization of antibody is detected:
Detect two kinds of monoclonal antibodies (Abnova company, #H00005710-AP11-2 that antibody selects the anti-human PSMD4 of mouse；
PTG company, #66179-1-Ig) detectable concentration selection (2.0 μ g/ml, 1 μ g/ml, 0.5 μ g/ml, 0.25 μ g/ml, 0.1 μ g/
ml).It is detected according to the operating procedure in embodiment 1, known various concentration PSMD4 protein standard substance is added, selection is empty
White group of OD value is minimum, and the conduct optimum detection antibody and work that the linear relationship between OD value and standard protein concentration is the closest
Make concentration.The 0.5 μ g/ml of working concentration of PSMD4 (PTG company) is best compatibility.
4, the optimization of ELIAS secondary antibody:
Detecting antibody is mouse anti human PSMD4 monoclonal antibody (Abnova company, #H00005710-AP11-2；PTG company, #
IgG (the Jackson company #715-005-150 that ELIAS secondary antibody is goat anti-mouse HRP label is chosen when 66179-1-Ig)；PTG
Company #SA00001-1)；Antibody extension rate selects 1:1000,1:2000,1:3000,1:5000,1:10000.According to implementation
Operating procedure in example 1 is detected, and known various concentration PSMD4 protein standard substance is added, and selects blank group OD value minimum,
And the best ELIAS secondary antibody of conduct and diluted concentration that the linear relationship between OD value and standard protein concentration is the closest.Goat is anti-
The IgG antibody compatibility of mouse HRP label is best (PTG company #SA00001-1), and diluted concentration is that 1:5000 is diluted to most preferably.
5, the optimization of sample diluting liquid:
Sample diluting liquid selects 1 × PBS (pH:7.4) or contains 0.1%BSA when patients serum's pattern detection, and 0.5%
1 × TBS solution of Tween-20, is detected according to the operating procedure in embodiment 1, and known various concentration PSMD4 is added
Protein standard substance selects the linear relationship between OD value and standard protein concentration the closest and dilutes as best serum sample
Liquid.Best serum sample dilution is 1 × PBS (pH:7.4) solution.
Embodiment 3:ELISA kit detects PSMD4 fusion protein
It by PSMD4 recombinant protein (PTG#ag6691) doubling dilution, is diluted since 10ng/ml, serial dilution 7 dense
Degree, 10ng/ml, 3ng/ml, 1ng/ml, 0.3ng/ml, 0.1ng/ml, 0.03ng/ml, each sample of 0.01ng/ml are arranged 2
Multiple holes, 100 holes μ l/, according to the step of embodiment 1 repeat detection 5 times, it is as a result similar, as shown in Figure 1, PSMD4 recombinant protein with
The reaction of antibody has good concentration-dependent relation, and at apparent linear relationship.
The specificity and sensitivity assessment of the ELISA kit of embodiment 4:PSMD4 albumen
1, specific test
50 normal populations, 26 lung cancer patient blood serum samples, 41 stomaches are detected with the ELISA method that embodiment 1 is established
(healthy population is selected from the court employee physical examination sample for carninomatosis human serum sample and 39 Patients with Colorectal Cancer serum samples；Lung cancer,
Gastric cancer and colorectal cancer sample standard deviation derive from Shanghai City Long March hospital laboratory, through pathological diagnosis be primary lung cancer, gastric cancer and
Colorectal cancer), using PSMD4 recombinant protein as positive control, standard is drawn according to gained standard items OD value and corresponding concentration
Curve calculates the serological levels of PSMD4 albumen in each crowd.
As shown in Fig. 2, serological levels (Mean178.99pg/mL, the Range0- of lung cancer group crowd's PSMD4 albumen
It 645.95pg/ml) is apparently higher than normal population (Mean98.84pg/mL, Range0-164.14pg/ml), difference has statistics
Meaning (p=0.002).Prompt PSMD4 has certain specificity in the serodiagnosis of lung cancer.
As shown in figure 3, serological levels (Mean193.99pg/mL, the Range20.82- of gastric cancer group crowd's PSMD4 albumen
It 586.43pg/ml) is apparently higher than normal population (Mean98.84pg/mL, Range0-164.14pg/ml), difference has statistics
Meaning (p ﹤ 0.001).Prompt PSMD4 has certain specificity in the serodiagnosis of gastric cancer.
As shown in figure 4, colorectal cancer group crowd's PSMD4 albumen serological levels (Mean258.01pg/mL,
Range20.76-1010.38pg/ml it) is apparently higher than normal population (Mean98.84pg/mL, Range0-164.14pg/ml),
Difference is statistically significant (p ﹤ 0.001).Prompt PSMD4 has certain specificity in the serodiagnosis of colorectal cancer.
(Mean- mean value；Range- extreme value range)
2, sensitivity tests
PSMD4 recombinant protein (1ug/ul) is made into continuous doubling dilution with 1 × PBS, 2 multiple holes are set, obtain standard egg
The mean OD value and blank group OD value of white each dilution point have the highest extension rate of statistical difference.
PSMD4 fusion protein dilution 108OD value and blank group OD value after times still have statistical difference, show detectable
PSMD4 albumen minimum concentration is 10pg/ml.
The basic principles, main features and advantages of the present invention have been shown and described above.The technology of the industry
Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the above embodiments and description only describe this
The principle of invention, various changes and improvements may be made to the invention without departing from the spirit and scope of the present invention, these changes
Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and its
Equivalent defines.
Claims (7)
1. a kind of ELISA detection kit of PSMD4 albumen is in preparation malignant tumour serodiagnosis or kit for screening
Using, which is characterized in that the malignant tumour is gastric cancer.
2. a kind of ELISA detection kit of PSMD4 albumen according to claim 1 is examined in preparation malignant tumour serology
Application in disconnected or kit for screening, which is characterized in that the ELISA detection kit of the PSMD4 albumen includes:
(A) it is coated with the ELISA ELISA Plate of PSMD4 antibody, the coated antibody is the polyclonal antibody of rabbit-anti people PSMD4；
(B) antibody for detecting PSMD4 antigen is the monoclonal antibody of mouse anti human PSMD4；
(C) ELIAS secondary antibody, for the goat anti-mouse IgG of HRP label；
(D) standard protein is recombined human PSMD4 fusion protein；
(E) sample diluting liquid, coating buffer, confining liquid, ELISA ELISA Plate cleaning solution, antibody diluent, developing solution and termination
Liquid.
3. a kind of ELISA detection kit of PSMD4 albumen according to claim 2 is examined in preparation malignant tumour serology
Application in disconnected or kit for screening, which is characterized in that the ELISA ELISA Plate is the ELISA ELISA Plate in 96 holes.
4. a kind of ELISA detection kit of PSMD4 albumen according to claim 2 or 3 is in preparation malignant tumour serum
Learn the application in diagnosis or kit for screening, which is characterized in that in the ELISA detection kit,
It is coated with buffer, is 1 × PBS, pH:7.4；
Confining liquid, for containing 3% bovine serum albumin(BSA)+PBS solution confining liquid；
ELISA ELISA Plate cleaning solution, for 1 × PBS solution containing 0.05%Tween-20；
Antibody diluent is 1 × PBS；
Developing solution is 3,3 ', 5,5 '-tetramethyl benzidines；
Terminate liquid is the sulfuric acid solution of 2mol/L.
5. a kind of ELISA detection kit of PSMD4 albumen according to claim 2 or 3 is in preparation malignant tumour serum
Learn the application in diagnosis or kit for screening, which is characterized in that in the ELISA detection kit, when sample is serum,
Sample diluting liquid is 1 × PBS, pH:7.4.
6. a kind of ELISA detection kit of PSMD4 albumen according to claim 2 or 3 is in preparation malignant tumour serum
Learn the application in diagnosis or kit for screening, which is characterized in that method of the kit for serodiagnosis or screening
The following steps are included:
Preparation detection ELISA ELISA Plate: the polyclonal antibody of rabbit-anti people PSMD4 is diluted to 2.0ug/ml by coating buffer；
Sample is diluted with sample diluting liquid with 1:1, is loaded into ELISA ELISA Plate hole, is incubated at room temperature 1--2 hours；
Recombined human PSMD4 fusion protein is diluted to various concentration gradient with sample diluting liquid, is loaded into ELISA ELISA Plate hole,
Incubation at room temperature 1--2 hours；The liquid in each hole is got rid of, ELISA ELISA Plate cleaning solution is added, is washed 3-5 times, drying；
The antibody that will test PSMD4 antigen is diluted to 0.5 μ g/ml, adds in ELISA ELISA Plate hole, is incubated at room temperature 1--2 hours；
The liquid in each hole is got rid of, ELISA ELISA Plate cleaning solution is added, is washed 3-5 times, drying；
The goat anti-mouse IgG of HRP label is added, is incubated at room temperature 1--2 hours；The liquid in each hole is got rid of, ELISA enzyme is added
Target cleaning solution washs 6-8 times, drying；
Developing solution is added, room temperature is protected from light incubation 10-20 minutes, and terminate liquid is added and terminates reaction；450nm measures OD in microplate reader
Value.
7. a kind of ELISA detection kit of PSMD4 albumen according to claim 2 or 3 is in preparation malignant tumour serum
Learn the application in diagnosis or kit for screening, which is characterized in that method of the kit for serodiagnosis or screening
Include:
Before detection, prepare detection ELISA ELISA Plate first: the polyclonal antibody of rabbit-anti people PSMD4 is diluted to by coating buffer
100 μ l are added in 2.0ug/ml in every hole of ELISA ELISA Plate, and sealing plate is placed on 4 DEG C of overnight incubations, spare；
Serum sample is diluted with 1 × PBS of sample diluting liquid, pH:7.4 with 1:1, is loaded with the volume in 100 holes μ l/, incubation at room temperature 2
Hour；PSMD4 recombinant protein is diluted to various concentration gradient with sample diluting liquid, is loaded with the volume in 100 holes μ l/, and room temperature is incubated
It educates 2 hours；The liquid in each hole is got rid of, ELISA ELISA Plate cleaning solution is added, every 300 μ l of hole is washed 3-5 times, drying；It will
PSMD4 detection antibody is diluted to 0.5 μ g/ml, every 100 μ l of hole, is incubated at room temperature 30 minutes 1 hour；The liquid in each hole is got rid of, is added
Enter ELISA ELISA Plate cleaning solution, every 300 μ l of hole is washed 3-5 times, drying；The goat anti-mouse IgG of HRP label, every hole is added
100 μ l are incubated at room temperature 1 hour；The liquid in each hole is got rid of, ELISA ELISA Plate cleaning solution is added, every 300 μ l of hole washs 6-8
It is secondary, drying；TMB, every 100 μ l of hole is added, room temperature is protected from light incubation 10-20 minutes, and 2mol/L sulfuric acid solution is added and terminates reaction, often
50 μ l of hole；450nm measures OD value in microplate reader.
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| PCT/CN2016/111720 WO2017107974A1 (en) | 2015-12-23 | 2016-12-23 | Detection test kit for serum psmd4 proteins and detection method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106885908B (en) * | 2015-12-23 | 2019-05-07 | 中国人民解放军第二军医大学 | The detection kit and its detection method of blood-serum P SMD4 albumen and application |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005094422A2 (en) * | 2004-02-20 | 2005-10-13 | Kevin Jon Williams | Angiocidin fragments and uses thereof in clinical assays for cancer and other diseases |
| CN1852974A (en) * | 2003-06-09 | 2006-10-25 | 密歇根大学董事会 | Compositions and methods for treating and diagnosing cancer |
| CN101441222A (en) * | 2008-12-23 | 2009-05-27 | 北京德宝群兴科技有限公司 | Method for detecting Beta accompany glycinin and specific antibody and reagent kit thereof |
| CN103091493A (en) * | 2013-01-08 | 2013-05-08 | 中国人民解放军第二军医大学 | Application of PSMD4 protein in preparation of liver cancer prognostic evaluation kit |
| CN105203760A (en) * | 2015-07-24 | 2015-12-30 | 中国人民解放军第二军医大学 | PSMD4 protein ELISA detection kit as well as detection method and application thereof |
| WO2017107974A1 (en) * | 2015-12-23 | 2017-06-29 | 中国人民解放军第二军医大学 | Detection test kit for serum psmd4 proteins and detection method and application thereof |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1109900A1 (en) * | 1999-06-21 | 2001-06-27 | Inkine Pharmaceutical Company Inc. | Angiocidin: a cys-ser-val-thr-cys-gly specific tumor cell adhesion receptor |
| US7655411B2 (en) * | 2002-08-23 | 2010-02-02 | W2 Holdings, Inc. | Thrombospondin fragments and binding agents in the detection, diagnosis and evaluation of cancer |
| CA2567350C (en) * | 2004-05-21 | 2018-06-12 | John D. Shaughnessy | Use of gene expression profiling to predict survival in cancer patient |
| US20080274911A1 (en) * | 2006-11-07 | 2008-11-06 | Burington Bart E | Gene expression profiling based identification of genomic signature of high-risk multiple myeloma and uses thereof |
| EP2115158A4 (en) * | 2006-12-27 | 2009-12-30 | Snu R&Db Foundation | Data processing, analysis method of gene expression data to identify endogenous reference genes |
| JP2009050189A (en) * | 2007-08-24 | 2009-03-12 | Sumitomo Bakelite Co Ltd | Method for predicting effectiveness of anti-cancer agent |
| AR090263A1 (en) * | 2012-03-08 | 2014-10-29 | Hoffmann La Roche | COMBINED ANTIBODY THERAPY AGAINST HUMAN CSF-1R AND USES OF THE SAME |
-
2015
- 2015-12-23 CN CN201510979741.3A patent/CN106885908B/en active Active
-
2016
- 2016-12-23 WO PCT/CN2016/111720 patent/WO2017107974A1/en active Application Filing
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1852974A (en) * | 2003-06-09 | 2006-10-25 | 密歇根大学董事会 | Compositions and methods for treating and diagnosing cancer |
| WO2005094422A2 (en) * | 2004-02-20 | 2005-10-13 | Kevin Jon Williams | Angiocidin fragments and uses thereof in clinical assays for cancer and other diseases |
| CN101441222A (en) * | 2008-12-23 | 2009-05-27 | 北京德宝群兴科技有限公司 | Method for detecting Beta accompany glycinin and specific antibody and reagent kit thereof |
| CN103091493A (en) * | 2013-01-08 | 2013-05-08 | 中国人民解放军第二军医大学 | Application of PSMD4 protein in preparation of liver cancer prognostic evaluation kit |
| CN105203760A (en) * | 2015-07-24 | 2015-12-30 | 中国人民解放军第二军医大学 | PSMD4 protein ELISA detection kit as well as detection method and application thereof |
| WO2017107974A1 (en) * | 2015-12-23 | 2017-06-29 | 中国人民解放军第二军医大学 | Detection test kit for serum psmd4 proteins and detection method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| Clinical significance of serum angiocidin levels in hepatocellular carcinoma;Sabherwal,Y等;《Cancer letters》;20070618;第251卷(第1期);28-35 |
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