CN102735846A - Chemiluminescence immunodetection kit and detection method for ovarian cancer tumor marker HE4 - Google Patents
Chemiluminescence immunodetection kit and detection method for ovarian cancer tumor marker HE4 Download PDFInfo
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- CN102735846A CN102735846A CN2012101987399A CN201210198739A CN102735846A CN 102735846 A CN102735846 A CN 102735846A CN 2012101987399 A CN2012101987399 A CN 2012101987399A CN 201210198739 A CN201210198739 A CN 201210198739A CN 102735846 A CN102735846 A CN 102735846A
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Abstract
The invention relates to the field of biological and medical detection technologies, specifically discloses a chemiluminescence immunodetection kit for the ovarian cancer tumor marker HE4 and also discloses a method of using the chemiluminescence immunodetection kit for the ovarian cancer tumor marker HE4 for detecting the ovarian cancer tumor marker HE4. The chemiluminescence immunodetection kit comprises a microplate coated by 9F3 antibodies, 10 E1 antibodies labeled by alkaline phosphatase, an analysis buffer solution, substrate operating fluid, a luminescent substrate, washing liquid and a quality control. The chemiluminescence immunodetection kit for the ovarian cancer tumor marker HE4 provided in the invention has a linear range of 0.5 ng/ml to 1200 ng/ml and a detection limit of 0.2 ng/ml, can be used for clinical auxiliary diagnosis, curative effect observation and prognosis of ovarian cancers and has important significance to treatment and prevention of ovarian cancer tumors.
Description
Technical field
The present invention relates to biology and technical field of medical detection; Be specifically related to a kind of oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit, also relate to the method that adopts this oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit to detect oophoroma tumor marker HE4 simultaneously.
Background technology
Oophoroma morbidity concealment, poor prognosis is described to be one of the most fatal cancer.The early symptom of oophoroma is very not obvious, and the initial stage symptom is abdominal pain or expansion, stomach indigestion, is difficult for causing enough attention, and general ovarian cancer patients all reaches an advanced stage when finding, so cure rate is extremely low.Just can improve the cure rate of oophoroma greatly if can find oophoroma in early days, in order to reach this purpose, people have carried out number of research projects.
CA125 is a kind of diagnosis of ovarian cancer and one of the most responsive index of its recurrence of monitoring, but CA125 the patient fall ill often detect in early days less than, its clinical practice has certain limitation.Given this, experts and scholars both domestic and external are devoted to seek more efficiently mark, so that can make a definite diagnosis oophoroma in early days.
(Human Epididymis Protein 4 HE4) finds in people's epididymal cell people's epididymis secretory protein 4.1999, Schummer etc. found mRNA high expressed in ovarian cancer tissue of HE4, expressed or did not express and in normal tissues, hang down.2003, HE4 content also had than the normal person and obviously increases in the discovery ovarian cancer patients serum such as Hellstrom, and therefore, HE4 has obtained more deep research as a new mark of oophoroma.Confirmations such as Hellstrom are that the sensitivity that HE4 detects oophoroma is 60% under 100% the situation in specificity, and CA125 is merely 13% as the sensitivity of the marker detection of oophoroma, and particularly the HE4 advantage that detects early ovarian cancer more obviously is superior to CA125.
Summary of the invention
The purpose of this invention is to provide a kind of oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit.
The present invention also aims to provide a kind of method that adopts this oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit to detect oophoroma tumor marker HE4.
In order to realize above purpose; The technical scheme that the present invention adopted is: a kind of oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit, this kit comprise microwell plate, the 10E1 antibody of alkali phosphatase enzyme mark, analysis buffer, substrate working fluid, luminous substrate, cleansing solution, the quality-control product of 9F3 antibody sandwich.
Wherein, the preparation method of the microwell plate of said 9F3 antibody sandwich is: be diluted to 0.1 microgram/microlitre to 9F3 antibody with encapsulating damping fluid, make 9F3 antibody sandwich liquid; The 9F3 antibody sandwich liquid that in each hole of microwell plate, adds 50 microlitres then respectively encapsulated 2 hours in 37 ℃, used the normal saline flushing microwell plate then three times; And then in each hole of microwell plate, add the shrouding liquid of 100 microlitres respectively, in room temperature sealing 2 hours, use twice of normal saline flushing microwell plate then; Freeze drying; Make the microwell plate of 9F3 antibody sandwich, be sealed in the aluminium foil bag, 4 ℃ of preservations are subsequent use.
Further, the said preparation method who encapsulates damping fluid is: getting concentration is that 0.05Mol/L, pH value are 9.6 carbonate buffer solution, to wherein adding NaN
3, make and encapsulate damping fluid, encapsulate NaN in the damping fluid
3Mass percent concentration be 0.2%.
The preparation method of said shrouding liquid is: to concentration is that 0.01Mol/L, pH value are to add BSA (bovine serum albumin(BSA)) and NaN in 7.4 the PBS damping fluid
3, making shrouding liquid, the mass percent concentration of BSA is 1% in the shrouding liquid that makes, NaN
3Mass percent concentration be 0.2%.
The preparation method of the 10E1 antibody of said alkali phosphatase enzyme mark is: get 2 milligrams alkaline phosphatase, 4 milligrams 10E1 antibody, add then in the PBS damping fluid, stir down; The glutaraldehyde that in the PBS damping fluid, adds 0.05 milliliter again makes mixed solution, and the volume of said mixed solution is 0.5 milliliter; Continue at room temperature to stir 15 minutes, the lucifuge reaction is 4 hours then, in said mixed solution, adds monoethanolamine more afterwards; The concentration of monoethanolamine is 0.1Mol/L, stirring at room 2 hours, 4 ℃ of following PBS dialysed overnight; Mix with isopyknic glycerine afterwards, add NaN then
3, NaN
3Addition to make NaN
3Mass percent concentration be 0.1%, make the 10E1 antibody of alkali phosphatase enzyme mark, subsequent use in 4 ℃ of preservations.
The preparation method of said analysis buffer is: to concentration is that 0.01Mol/L, pH value are to add BSA and NaN in 8.0 the Tris-Hcl damping fluid
3, stir, make analysis buffer, the mass percent concentration of BSA is 1% in the said analysis buffer, NaN
3Mass percent concentration be 0.2%.
The preparation method of said substrate working fluid is: NaOH solution, the 100 microlitre concentration of getting 600 microlitre diethanolamine, 100 microlitre concentration and be 1Mol/L are the MgCl of 1Mol/L
2Solution, 100 microlitre mass percent concentrations are 10% NaN
3Solution, mixing, high pressure steam sterilization is processed substrate buffer solution; Measure the said substrate buffer solution of 800 microlitres; (the luminous substrate reinforcing agent is the supporting use product of luminous substrate in substrate buffer solution, to add 200 microlitre luminous substrate reinforcing agents then; Available from Bio-Rad company) and 50 microlitre luminous substrate; Mixing in sterilization container makes the substrate working fluid then.
Said luminous substrate is selected from any among AMPPD, CSPD, the CDP-Star.Luminous substrate is preferably CSPD.
The preparation method of said cleansing solution is: in concentration is that 0.01Mol/L, pH value are to add polysorbas20 and NaN in 7.4 the PBS damping fluid
3, stir, make cleansing solution, the mass percent concentration of polysorbas20 is 0.05% in the said cleansing solution, NaN
3Mass percent concentration be 0.2%.
Said quality-control product is the HE4 standard items.
When the microwell plate of preparation 9F3 antibody sandwich, microwell plate can be selected 24 holes, 48 holes or 96 orifice plates, preferably 96 orifice plates.
A kind of oophoroma tumor marker HE4 chemical luminous immune detection method adopts double antibody sandwich method, may further comprise the steps:
(1) testing sample and the HE4 standard solution with concentration gradient are added respectively in the different holes of microwell plate of 9F3 antibody sandwich; And then Xiang Kongzhong adds the 10E1 antibody of the alkali phosphatase enzyme mark that can combine with HE4; Add luminous substrate afterwards, detect, obtain testing result;
(2) do typical curve according to the luminous intensity values of the HE4 standard solution of measuring with concentration gradient;
(3) do comparison with the luminous intensity values of the testing sample measured and the luminous intensity values of typical curve, draw the content of HE4 in the testing sample.
Wherein, said testing sample is a human serum.
Chemiluminescence immune assay (Chemiluminescence Immunoassay; CLIA) be that immune response with highly sensitive chemical luminescent detecting technology and high selectivity combines, the check and analysis that are used for various antigens, haptens, antibody, hormone, enzyme, protein, vitamin, medicine etc. are technological.Be to exempt from, put an immunoassay of exempting from, growing up after the fluorescence, time-resolved fluorescence analysis continue enzyme.Chemiluminescence enzyme immunoassay (Chemiluminescent Enzyme Immunoassay; CLEIA) belong to EIA enzyme immunoassay; Just the substrate of enzyme reaction is a luminous agent, and operation steps and EIA enzyme immunoassay are identical, carry out immune response with the enzyme labeling bioactivator; Enzyme on the immune response compound remakes and is used for luminous substrate, carries out luminescence assays with the luminous signal analyzer.
Oophoroma tumor marker HE4 chemical luminous immune detection method provided by the invention; With the reorganization HE4 albumen bought as the antigen immune BALB/c mouse; The preparation monoclonal antibody is done pairing test, affinity test, cross reaction test respectively to the monoclonal antibody that obtains, and obtains two strain of hybridoma strain 10E1 and 9F3; The antibody capable pairing of hybridoma cell strain 10E1 and 9F3 secretion is used and is had high specific and high-affinity, and the affinity constant of 10E1 antibody reaches 10
-11M/L, the affinity constant of 9F3 antibody reaches 10
-12M/L; 10E1 antibody and 9F3 antibody can combine with HE4 different antigens determinant respectively; Through with the 10E1 antibody of alkaline phosphatase (ALP) mark as detecting antibody, be coated on the solid phase carrier as capture antibody with unlabelled 9F3 antibody, set up the double antibody sandwich method that detects HE4; The chemiluminescence immune assay of having realized oophoroma tumor marker HE4 detects, and judging for early diagnosis, observation of curative effect and the prognosis of oophoroma provides a kind of reliable detection method.
Oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit provided by the invention has the extraordinary range of linearity, and its range of linearity is 0.5 nanograms/milliliter～1200 nanograms/milliliter, detects to be limited to 0.2 nanograms/milliliter.Betweenrun precision＜8% of oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit provided by the invention, withinrun precision＜5%.Oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit provided by the invention can be used for clinical assistant diagnosis, observation of curative effect and the prognosis of oophoroma to be judged, to the treatment of oophoroma tumor with prevent significant.
Description of drawings
Fig. 1 is the SDS-PAGE electrophoretogram of the reorganization HE4 albumen of the embodiment of the invention 1 purchase;
The double-log canonical plotting that Fig. 2 obtains for the embodiment of the invention 2.
Embodiment
Through specific embodiment technical scheme of the present invention is elaborated below.
The oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit that present embodiment provides comprises microwell plate, the 10E1 antibody of alkali phosphatase enzyme mark, analysis buffer, substrate working fluid, luminous substrate, cleansing solution, the quality-control product of 9F3 antibody sandwich.
The preparation method of the microwell plate of 9F3 antibody sandwich is: be diluted to 0.1 microgram/microlitre to 9F3 antibody with encapsulating damping fluid, make 9F3 antibody sandwich liquid, in each hole of microwell plate, add the 9F3 antibody sandwich liquid of 50 microlitres then respectively; Encapsulated 2 hours in 37 ℃, use the normal saline flushing microwell plate then three times, and then in each hole of microwell plate, add the shrouding liquid of 100 microlitres respectively; In room temperature sealing 2 hours; Use normal saline flushing microwell plate twice then, freeze drying makes the microwell plate of 9F3 antibody sandwich; Be sealed in the aluminium foil bag, 4 ℃ of preservations are subsequent use.
Wherein, the preparation method who encapsulates damping fluid is: getting concentration is that 0.05Mol/L, pH value are 9.6 carbonate buffer solution, to wherein adding NaN
3, make and encapsulate damping fluid, encapsulate NaN in the damping fluid
3Mass percent concentration be 0.2%.
The preparation method of shrouding liquid is: to concentration is that 0.01Mol/L, pH value are to add BSA and NaN in 7.4 the PBS damping fluid
3, making shrouding liquid, the mass percent concentration of BSA is 1% in the shrouding liquid that makes, NaN
3Mass percent concentration be 0.2%.
The preparation method of the 10E1 antibody of alkali phosphatase enzyme mark is: get 2 milligrams alkaline phosphatase, 4 milligrams 10E1 antibody, add then in the PBS damping fluid, stir down; The glutaraldehyde that in the PBS damping fluid, adds 0.05 milliliter again makes mixed solution, and the volume of mixed solution is 0.5 milliliter; Continue at room temperature to stir 15 minutes, the lucifuge reaction is 4 hours then, in mixed solution, adds monoethanolamine afterwards again; The concentration of monoethanolamine is 0.1Mol/L, stirring at room 2 hours, 4 ℃ of following PBS dialysed overnight; Mix with isopyknic glycerine afterwards, and then add NaN
3, NaN
3Addition to make NaN
3Mass percent concentration reach 0.1%, make the 10E1 antibody of alkali phosphatase enzyme mark, subsequent use in 4 ℃ of preservations.
The preparation method of analysis buffer is: to concentration is that 0.01Mol/L, pH value are to add BSA and NaN in 8.0 the Tris-Hcl damping fluid
3, stir, make analysis buffer, the mass percent concentration of BSA is 1% in the analysis buffer, NaN
3Mass percent concentration be 0.2%.
The preparation method of substrate working fluid is: NaOH solution, the 100 microlitre concentration of getting 600 microlitre diethanolamine, 100 microlitre concentration and be 1Mol/L are the MgCl of 1Mol/L
2Solution, 100 microlitre mass percent concentrations are 10% NaN
3Solution, mixing, high pressure steam sterilization is processed substrate buffer solution; Measure 800 microlitre substrate buffer solutions; (this luminous substrate reinforcing agent is the supporting sell goods of luminous substrate CSPD in substrate buffer solution, to add 200 microlitre luminous substrate reinforcing agents then; Available from Bio-Rad company) and 50 microlitre CSPD (available from Bio-Rad company); Mixing in sterilization container makes the substrate working fluid then.
Luminous substrate is CSPD (available from a Bio-Rad company).
The preparation method of cleansing solution is: in concentration is that 0.01Mol/L, pH value are to add polysorbas20 and NaN in 7.4 the PBS damping fluid
3, stir, make cleansing solution, the mass percent concentration of polysorbas20 is 0.05% in the cleansing solution, NaN
3Mass percent concentration be 0.2%.
Quality-control product is the HE4 standard items.
Wherein, the preparation process of 9F3 antibody and 10E1 antibody is:
(1) mouse hybridoma cell prepares monoclonal antibody
The foundation of a, antibody cell strain
With conventional method BALB/c mouse is carried out immunity; Initial immunity with 100 microgram recombinant antigen HE4 albumen (available from Abnova company; The SDS-PAGE electrophoretogram of reorganization HE4 albumen is seen shown in Figure 1) with Freund's complete adjuvant fully emulsified after; Abdominal cavity or subcutaneous multi-point injection whenever mix the back injection at a distance from two weeks with 50 micrograms reorganization HE4 albumen later on incomplete Freund, immunity is four times altogether.Fusion of Cells first three day booster immunization once.From immunity beginning for the second time, the 7th day tail vein after each immunity gathered mouse blood, adopts indirect ELISA method to detect serum titer and reaches the above mouse of 1:50000, takes out the spleen of mouse, processes splenocyte suspension.Under the fusion of PEG,, and select nutrient culture media top sieve menu clone at HAT with the SP2/0 Fusion of Cells.Through the screening of 2-3 fusion, obtain the monoclonal hybridoma strain that anti-HE4 antibody is secreted in 26 strains altogether with the 3-5 wheel.
B, antibody pairing
Do antibody pairing test with the ELISA method, the result shows, 10E1 and 9F3, and 3A2 and 4E7 pairing are good.
(2) evaluation of monoclonal antibody
Hypotype classification: the hypotype of confirming monoclonal antibody with the mouse monoclonal antibody hypotype classification agent box of Sigma company through the Elisa method.The result shows that 10E1 and 3A2 are the IgG1 type, and 9F3 and 4E7 are the IgG2a type.
The mensuration of affinity: carry out relative affinity with indirect Elisa method and measure.HE4 with 1 mcg/ml encapsulates carrier board; With the AC is horizontal ordinate, and antibody is done serial dilution, to have the colour developing of HRP (horseradish peroxidase) sheep anti-mouse antibody; The OD value that detects with ELIASA is an ordinate; To be tending towards smooth OD value on the curve is 100%, finds its OD value and be 50% AC, and the affinity of the low more antibody of this concentration value is just high more.Convert through measured value, 10E1 reaches 10
-11M/L, 9F3 reaches 10
-12M/L, 3A2 reaches 10
-9M/L, 4E7 reaches 10
-10M/L.
Specificity analyses: with the HE4 approximate irrelevant protein of resulting monoclonal antibody with series, and the human serum that does not contain HE4 does test analysis, and the result shows that 10E1 and 9F3 all do not have and the cross reaction of irrelevant antigen, and 3A2 and 4E7 have cross reaction.
(3) mouse ascites prepares monoclonal antibody
With BALB/c mouse of 500 microlitre incomplete Freund sensitization, sensitization one all pneumoretroperitoneum injection hybridoma suspensions, cell number is 1 * 10
7, gather mouse ascites after 10 days, 12000 change high speed centrifugation, collect supernatant, carry out purifying with ProteinA, make 9F3 antibody and 10E1 antibody, and the Brodford method is measured AC, adds protein protective agent, in-70 ℃ of preservations.
The oophoroma tumor marker HE4 chemical luminous immune detection method that present embodiment provides, the kit that adopts the embodiment of the invention 1 to provide may further comprise the steps:
(1) in the PBS damping fluid, adds BSA, be mixed with the BSA mass percent concentration and be the PBS damping fluid of 1% BSA,, process reorganization HE4 protein standard substance solution with series concentration gradient with the reorganization HE4 albumen that this damping fluid dilution is bought;
(2) get the reorganization HE4 protein standard substance solution that 50 microlitre testing sample human serums and 50 microlitres have the series concentration gradient respectively, in the different holes of the microwell plate of adding 9F3 antibody sandwich, add 10E1 antibody working fluid (1:2000) 50 microlitres of ALP mark afterwards; Add luminous substrate CSPD afterwards; Concussion evenly, 37 ℃ of incubations 1 hour are washed 5 times with cleansing solution; Control is done on thieving paper; Add each 50 microlitre of substrate working fluid and analysis buffer again, the reaction of room temperature lucifuge was measured luminous intensity after 10 minutes on light-emitting appearance.Doing double-log to the concentration value of the HE4 standard items of variable concentrations (concentration unit is a nanograms/milliliter) and luminous intensity (unit the is RLU) data that record handles; Draw out the double-log typical curve; The double-log typical curve that obtains is seen shown in Figure 2; Wherein horizontal ordinate is a HE4 concentration logarithm value, and ordinate is the luminous intensity logarithm value.The concentration of HE4 can calculate by the double-log typical curve in the testing sample human serum.
Can find out that from the double-log typical curve Fig. 2 that records kit has the extraordinary range of linearity, the range of linearity is 0.5 nanograms/milliliter～1200 nanograms/milliliter, detects to be limited to 0.2 nanograms/milliliter.
Claims (9)
1. oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit; It is characterized in that, comprise microwell plate, the 10E1 antibody of alkali phosphatase enzyme mark, analysis buffer, substrate working fluid, luminous substrate, cleansing solution, the quality-control product of 9F3 antibody sandwich.
2. oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit according to claim 1; It is characterized in that the preparation method of said analysis buffer is: to concentration is that 0.01Mol/L, pH value are to add BSA and NaN in 8.0 the Tris-Hcl damping fluid
3, stir, make analysis buffer, the mass percent concentration of BSA is 1% in the said analysis buffer, NaN
3Mass percent concentration be 0.2%.
3. oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit according to claim 1; It is characterized in that the preparation method of said substrate working fluid is: NaOH solution, the 100 microlitre concentration of getting 600 microlitre diethanolamine, 100 microlitre concentration and be 1Mol/L are the MgCl of 1Mol/L
2Solution, 100 microlitre mass percent concentrations are 10% NaN
3Solution, mixing, high pressure steam sterilization is processed substrate buffer solution; Measure the said substrate buffer solution of 800 microlitres, in substrate buffer solution, add 200 microlitre luminous substrate reinforcing agents and 50 microlitre luminous substrate then, mixing in sterilization container makes the substrate working fluid then.
4. according to claim 1 or 3 described oophoroma tumor marker HE4 chemiluminescence immune detection reagent kits, it is characterized in that said luminous substrate is selected from any among AMPPD, CSPD, the CDP-Star.
5. oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit according to claim 4 is characterized in that said luminous substrate is CSPD.
6. oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit according to claim 1 is characterized in that the preparation method of said cleansing solution is: in concentration is that 0.01Mol/L, pH value are to add polysorbas20 and NaN in 7.4 the PBS damping fluid
3, stir, make cleansing solution, the mass percent concentration of polysorbas20 is 0.05% in the said cleansing solution, NaN
3Mass percent concentration be 0.2%.
7. oophoroma tumor marker HE4 chemiluminescence immune detection reagent kit according to claim 1 is characterized in that said quality-control product is the HE4 standard items.
8. an oophoroma tumor marker HE4 chemical luminous immune detection method is characterized in that, adopts double antibody sandwich method, may further comprise the steps:
(1) testing sample and the HE4 standard solution with concentration gradient are added respectively in the different holes of microwell plate of 9F3 antibody sandwich; And then Xiang Kongzhong adds the 10E1 antibody of the alkali phosphatase enzyme mark that can combine with HE4; Add luminous substrate afterwards, detect, obtain testing result;
(2) do typical curve according to the luminous intensity values of the HE4 standard solution of measuring with concentration gradient;
(3) do comparison with the luminous intensity values of the testing sample measured and the luminous intensity values of typical curve, draw the content of HE4 in the testing sample.
9. oophoroma tumor marker HE4 chemical luminous immune detection method according to claim 8 is characterized in that said testing sample is a human serum.
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| CN103389381A (en) * | 2013-07-19 | 2013-11-13 | 武汉生之源生物科技有限公司 | Human epididymal secretory protein E4 chemiluminescence detection kit and preparation method thereof |
| CN104237532A (en) * | 2014-09-10 | 2014-12-24 | 广州市达瑞抗体工程技术有限公司 | Quantitative determination kit for human epididymis secretory protein 4 |
| CN105606596A (en) * | 2016-01-26 | 2016-05-25 | 河南生生医疗器械有限公司 | Kit for chemiluminiscent immunodetection of brain fatty acid binding protein and preparation method thereof |
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| CN106370654A (en) * | 2016-08-25 | 2017-02-01 | 上海康元科技发展有限公司 | Solid state linear continuous gradient measurement method |
| CN106483293A (en) * | 2016-10-13 | 2017-03-08 | 广州华弘生物科技有限公司 | He4a clinical immunization detection kit and preparation method thereof |
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| CN103389381A (en) * | 2013-07-19 | 2013-11-13 | 武汉生之源生物科技有限公司 | Human epididymal secretory protein E4 chemiluminescence detection kit and preparation method thereof |
| CN103389381B (en) * | 2013-07-19 | 2015-07-15 | 武汉生之源生物科技有限公司 | Human epididymal secretory protein E4 chemiluminescence detection kit and preparation method thereof |
| CN104237532A (en) * | 2014-09-10 | 2014-12-24 | 广州市达瑞抗体工程技术有限公司 | Quantitative determination kit for human epididymis secretory protein 4 |
| CN105606596A (en) * | 2016-01-26 | 2016-05-25 | 河南生生医疗器械有限公司 | Kit for chemiluminiscent immunodetection of brain fatty acid binding protein and preparation method thereof |
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| CN111298138B (en) * | 2020-04-15 | 2022-04-26 | 徐州医科大学 | Ovarian cancer diagnosis and treatment integrated nano probe BSA-Gd2O3/PTX@Anti-HE4 mAb |
| CN111781364A (en) * | 2020-08-25 | 2020-10-16 | 北京信诺卫康科技有限公司 | Wnt7a and HE4 combined as early ovarian cancer biomarker and kit |
| CN111912987A (en) * | 2020-08-25 | 2020-11-10 | 北京信诺卫康科技有限公司 | Combination of FGF18 and HE4 as early ovarian cancer biomarker and kit |
| CN112979814A (en) * | 2021-04-18 | 2021-06-18 | 深圳市国创纳米抗体技术有限公司 | anti-HE 4 nano antibody 3C8 and application thereof |
| CN112979815A (en) * | 2021-04-18 | 2021-06-18 | 深圳市国创纳米抗体技术有限公司 | anti-HE 4 nano antibody 1G8 and application thereof |
| CN112979815B (en) * | 2021-04-18 | 2022-04-22 | 深圳市国创纳米抗体技术有限公司 | anti-HE 4 nano antibody 1G8 and application thereof |
| CN112979814B (en) * | 2021-04-18 | 2022-04-22 | 深圳市国创纳米抗体技术有限公司 | anti-HE 4 nano antibody 3C8 and application thereof |
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