CN101377509A - III type precollagen N end peptide chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof - Google Patents
III type precollagen N end peptide chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof Download PDFInfo
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- CN101377509A CN101377509A CN 200810102020 CN200810102020A CN101377509A CN 101377509 A CN101377509 A CN 101377509A CN 200810102020 CN200810102020 CN 200810102020 CN 200810102020 A CN200810102020 A CN 200810102020A CN 101377509 A CN101377509 A CN 101377509A
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Abstract
The invention relates to the medical field of immunoassay, more specially, the invention provides a chemiluminescent immunoassay quantitative detection kit for III-type procollagen N-terminal peptide (PIIINP) and a preparation method thereof. The kit of the invention comprises 1) III-type procollagen N-terminal peptide calibrators, 2) solid-phase vectors which are coated with III-type procollagen N-terminal peptide monoclonal antibodies, 3) enzyme markers for III-type procollagen N-terminal peptide monoclonal antibodies, 4) chemiluminescent substrates and 5) concentrated washing solution. Further, the preparation method of the kit according to the invention comprises the following steps: 1) preparing the III-type procollagen N-terminal peptide calibrators with pure III-type procollagen N-terminal peptide, 2) coating the vectors with III-type procollagen N-terminal peptide monoclonal antibodies, 3) marking the III-type procollagen N-terminal peptide monoclonal antibodies with the enzyme, 4) preparing the chemiluminescent substrates, 5) preparing the concentrated washing solution, 6) packaging the III-type procollagen N-terminal peptide calibrators, the enzyme markers, the chemiluminescent substrates and the concentrated washing solution and 7) assembling finished products. The kit of the invention has the advantages of simplicity, convenience, rapidness, sensitivity, stability and the like.
Description
Technical field
The present invention relates to the immunoassay medical domain, particularly, the invention provides a kind of III procollagen type N end peptide chemiluminescence immune analysis quantitative determination reagent kit and preparation method thereof.
Background technology
Liver fibrosis is the only stage which must be passed bies of various chronic liver diseases to the cirrhosis development.Cirrhosis is a class serious threat human life and healthy malignant disease.Diagnosing early and treating liver fibrosis is one of effective means of antagonism cirrhosis, and therefore, the early diagnosis of liver fibrosis is very important.
At present, the blood serum designated object of clinical diagnosing liver fibrosis commonly used has III procollagen type N end peptide, IV Collagen Type VI, III procollagen type N end peptide, prolyl hydroxylase, Laminin ELISA, Laminin ELISA-P1 fragment, TGF-β 1 etc., and III procollagen type N end peptide commonly used, IV Collagen Type VI, III procollagen type N end peptide, Laminin ELISA are as the mark of fine four the diagnosing liver fibrosis degree of liver.Now, the liver fibrosis diagnosis kit adopts radioimmunoassay method more, and this method is harmful, easily pollutes.General detection method is difficult to it is carried out quantitatively accurate, and III procollagen type N end peptide is one of liver fibrosis mark of using always, and to degree of hepatic fibrosis, particularly early diagnosis has higher-value.Therefore, how can measure III procollagen type N end peptide level and the common diagnosing liver fibrosis of other mark degree of examining effectively is the problem that people pay close attention to and wish to solve always.
The research of labelled immune analytical technology and application development are rapid over past ten years, have been widely used in each field of biomedical fundamental research and clinical disease diagnosis.Wherein the technical matters maturation has advance and practical, and what be easy to promote mainly contains: methods such as radiommunoassay, enzyme-linked immuno assay, time resolved fluoro-immunoassay and chemiluminescence immune assay.The basic theories of these ultramicron detection techniques is identical substantially, but used tracer agent and the signal that sent have nothing in common with each other.According to lot of experiment results and clinical practice data,, be followed successively by: chemiluminescence immune assay, time resolved fluoro-immunoassay, enzyme-linked immuno assay and radiommunoassay from practicality, stability, accuracy and development prospect.III procollagen type N end peptide detection method mainly contains radio immunoassay at present, and enzyme linked immunological adsorption chromatography and time-resolved fluorescence immunoassay method etc. are the most universal with enzyme linked immunological adsorption chromatography method.There are many shortcomings in radio immunoassay, as complicated operation, measurement result instability, reagent holding time weak point, radioactive contamination, instrument costliness etc.; Time-resolved fluorescence immunoassay method need carry out mark with rare metal; The enzyme-linked immuno assay skill has overcome radioactive contamination, uses extensively, but owing to have the enzyme instability, the photometric measurement narrow range, sensitivity is not high, is difficult to weak points such as quantitative, has been subjected to the restriction of using; Chemiluminescence immune assay has highly sensitive, high specificity, and the range of linearity is wide, detect advantages such as quick, and agents useful for same toxicity is low, "dead" pollution, and good stable is arranged; Widely applicable, be widely used for the immune detection of antigen, haptens, antibody.
Chemiluminescence immunoassay is a kind of than elder generation and then effective method, yet chemiluminescence immunoassay technology still is in the starting developing stage in the application facet of III procollagen type N end peptide immunoassay product at present.On the one hand, the industry application need solves the high efficiency and the stability problem of finished product.On the other hand, chemical luminescence immune analysis reagent box of the prior art is the luminous measuring system of closed full-automatic chemical, need the expensive luminous measuring instrument of full-automatic chemical, promote the use of, can't be widely used in clinical diagnosis and research work thereby limited.Kit of the present invention can be applicable to the open luminous measuring instrument of semi-automatic chemistry, also can be used for full automatic measuring system, can realize fast detection the in enormous quantities, and use cost is low, easier applying.
Summary of the invention
The present invention has solved the problems referred to above simultaneously, soon chemiluminescence is learned effectively with the immunoassay of III procollagen type N end peptide and is combined, a kind of kit that can easy, quick, sensitive, stably detect III procollagen type N end peptide is provided, and this kit is suitable for applying effectively on industry.
The purpose of this invention is to provide a kind of III procollagen type N end peptide chemiluminescence immune analysis quantitative determination reagent kit and preparation method thereof.
Kit according to the present invention comprises: 1) III procollagen type N end peptide calibration object; 2) be coated with the solid phase carrier that III procollagen type N holds the monoclonal antibody of peptide; 3) monoclonal antibody of the III procollagen type N of enzyme labeling end peptide; The chemical luminous substrate that above-mentioned enzyme acted on; And 5) concentrated cleaning solution.
Further, the method for preparing the mentioned reagent box according to the present invention may further comprise the steps:
1) with the pure product preparation of III procollagen type N end peptide III procollagen type N end peptide calibration object;
2) monoclonal antibody of holding peptide with enzyme labeling III procollagen type N;
3) with the monoclonal antibody bag suppressed by vector of III procollagen type N end peptide;
4) preparation chemical luminous substrate;
5) preparation concentrated cleaning solution;
6) monoclonal antibody of the III procollagen type N end peptide of the above-mentioned III procollagen type of packing N end peptide calibration object, enzyme labeling, chemical luminous substrate and the concentrated cleaning solution that this enzyme acted on; And
7) be assembled into finished product.
Above-mentioned according to kit of the present invention and preparation method thereof in, described carrier is solid phase carrier, preferred microporous plate, plastic bead, plastic tube or magnetic-particle.Described enzyme can be alkaline phosphatase or horseradish peroxidase.Described chemical luminous substrate can be 1,2-two oxidative ethane analog derivatives, luminol or different luminol, wherein said 1,2-two oxidative ethane analog derivatives can be (diamantane)-1,2-two oxidative ethanes, 3-(2 '-the spiral diamantane)-4-methoxyl-4-(3 " the phosphorus acyloxy) phenyl-1,2-two oxidative ethanes (AMPPD), CSPD or CDP-Star.
In the method according to the invention, the step 3) of wherein said bag suppressed by vector may further comprise the steps:
1) bag quilt
The bag for preparing is cushioned liquid mixes, and the gained mixed liquor is carried on the solid phase carrier with the monoclonal antibody of III procollagen type N end peptide;
2) wash above-mentioned carrier with physiological saline; And
3) sealing
The confining liquid of preparation is sealed solid phase carrier after the above-mentioned washing.
Concrete mentioned reagent box can comprise III procollagen type N end peptide calibration object, antibody sandwich plate, enzyme labeling thing and chemical luminous substrate liquid, 20 times of concentrated cleaning solutions etc.Wherein, described III procollagen type N end peptide calibration object is a standard level, and micropore lath, enzyme labeling III procollagen type N end peptide monoclonal antibody that purity is not less than 90%, the antibody sandwich plate is 48 or 96 holes are that coupling alkaline phosphatase, chemical luminous substrate liquid are that AMPPD, concentrated cleaning solution are the Tris-HCl washing lotion.
The present invention's " III procollagen type N end peptide chemiluminescence immune analysis quantitative determination reagent kit " detection by quantitative very single-mindedly goes out the content that patient III procollagen type N holds peptide, can treat the effect of liver fibrosis patient medicine and the variation of the state of an illness thereof according to how many judgements of III procollagen type N end peptide content.It has advantages such as easy, quick, sensitive, stable.Every index that this III procollagen type N end peptide is measured kit (chemoluminescence method) all meets or exceeds enzyme-linked immunosorbent assay.And, according to detection system of the present invention is open-sky technique, easy to be quick, does not need the expensive luminous measuring instrument of full-automatic chemical, be particularly suitable for vast middle and small hospital and promote the use of, for clinical diagnosis and research work provide a kind of very valuable detection means.According to kit of the present invention, the III procollagen type N end peptide protein fragment of the III procollagen type N end peptide monoclonal antibody of bag quilt and sample forms " double-antibody sandwich " structure on the III procollagen type N end peptide monoclonal antibody of enzyme labeling and the carrier, therefore " double-antibody sandwich single stage method " reaction pattern of adopting of the present invention had not only effectively utilized the chemiluminescence principle, but also had guaranteed the sensitivity that detects.In addition, this pattern also is convenient to operation and is produced.
What kit of the present invention was used is the enzymatic luminous substrate, by the chromogenic substrate in the light signal replacement enzyme-linked immuno assay that detects the luminous substrate generation, thereby have a specificity equal with enzyme-linked immuno assay, and sensitivity improves greatly, ratio Enzyme Linked Immunoadsorbent Assay sensitivity now improves greatly, and the diagnosis that can be liver fibrosis provides more special, quick, reliable foundation.
Description of drawings
Fig. 1 is the calibration object linear graph in the prepared kit of embodiment 1.
The correlativity of Fig. 2 kit of the present invention and PIIINP enzyme-linked immunologic detecting kit.
Embodiment
One, enzyme labelled antibody preparation
III procollagen type N end peptide monoclonal antibody is fully dialysed to PBS with glutaraldehyde method and alkaline phosphatase coupling, adds equal-volume glycerine, preserves below-20 ℃.
Two, the preparation of III procollagen type N end peptide calibration object
With the pure product preparation of III procollagen type N end peptide, totally 6 bottles of packing 0,6.2,12.5,25,50,100ng/ml.
Three, enzyme labelled antibody concentration is selected
Adopt the square formation method to select the working concentration of enzyme labelled antibody greater than 1:6000.
Four, the preparation of solid-phase coating plate
(1) bag quilt
Trisodium citrate 2.92g
Citric acid 1.78g
Distilled water 400ml
Behind the dissolving mixing, adjust PH to 4.8, add an amount of III procollagen type N end peptide monoclonal antibody mixing, add then in each hole of microwell plate, every hole 110 μ L, 4 ℃ are spent the night.
(2) washing: it is inferior to give a baby a bath on the third day after its birth with physiological saline.
(3) sealing
NaH
2PO
4·2H
2O 0.2g
Na
2HPO
4·12H
2O 2.9g
BSA 10g
Proclin300 1ml
Distilled water is settled to 1000ml
The mentioned reagent weighing is put into clean container well, add the distilled water constant volume, the dissolving mixing, measuring the pH value is 7.0.
Every hole adds confining liquid 300 μ L respectively, and room temperature was placed 3 hours.Get rid of confining liquid, on thieving paper, pat dry.Room temperature removal moisture drying 24 hours.Carry out vacuum sealing bag immediately, put into one piece of drying agent in every bag.Place behind the envelope and checked not have gas leakage in 15 minutes, if there is gas leakage to need envelope again.2～8 ℃ of preservations of labeling postposition.
Five, chemical luminous substrate liquid
Tris 24g
NaCl 160g
KCl 4g
HCl 15mL
Distilled water 1000mL
Proclin 300 1mL
AMPPD 200ml
Dissolving and mixing.
Six, 20 times of concentrated cleaning solutions
Tris 24g
NaCl 160g
KCl 4g
HCl 15ml
Deionized water 1000ml
Adjust PH to 7.4
Seven, semi-manufacture and finished product are formed
The packing of above-mentioned steps products obtained therefrom is semi-manufacture.Extract three parts of process specificitys, accuracy, sensitivity and stable assay approvals out and just can be assembled into III procollagen type N end peptide quantitative determination reagent kit (chemoluminescence method).Be assembled into also need inspect by random samples behind the kit and just can dispatch from the factory after qualified.
To sum up, in research process of the present invention, the present inventor has at first carried out screening experiment and Quality Identification to used starting material, comprises the luminous intensity of activity, chemical luminous substrate of the absorption property of activity, carrier (as lighttight white microwell plate) of labelled antibody and coated antibody and variation size, ALP and luminous duration etc.Then method for coating is studied, be cushioned liquid and protect liquid to experimentize, select optimal bag and be cushioned liquid and protection liquid, tested by concentration by the different bags of antibody and find best concentration conditions with different bags.Mark for ALP can have diverse ways, by explore repeatedly and the contrast experiment finally found easy, productive rate is high, cost is low, the reliable quality labeling method, and different enzyme dilutions has been carried out long-term investigation tested, made and can make the activity stabilized dilution of the long-term maintenance of enzyme labeling thing.
The present inventor has also carried out experimental study to the reaction pattern and the reaction conditions of kit, finally determined double-antibody sandwich single stage method reaction pattern, and the influence of experimental result is tested with regard to the different reaction time, determine the optimal reaction time.By the influence experiment to measured value shows to the mensuration of luminous duration of chemical luminous substrate liquid and different fluorescent lifetime: be measured as the best between 30-90 minute after adding chemical luminous substrate liquid, its result is also comparatively accurate.
Embodiment 2～3 preparations III procollagen type N end of the present invention peptide quantitative determination reagent kit
Divided by horseradish peroxidase mark III procollagen type N end peptide monoclonal antibody, with luminol (luminol) system as chemical luminous substrate, this chemical luminous substrate comprises A liquid and B liquid, based on the described chemical luminous substrate A of 1000mL liquid, comprise 1.7716g luminol, 0.051g 4-xenol, 0.012g 4-iodobenzene boric acid, 11.4g boric acid, 4.9g borax, its pH value is 8.0～10.0; Based on the described chemical luminous substrate B of 1000mL liquid, comprise 0.329g urea peroxide, 1ml Tween20,51.58g Na
2HPO
412H
2O, 8.74gNaH
2PO
42H
2O, its pH value is 7.0～7.6, respectively with plastic bead, plastic tube as the pH value of solid phase carrier, coating buffer be 4.5, the pH value of confining liquid is outside 7.5, all the other all prepare III procollagen type N end peptide quantitative determination reagent kit with the method identical with embodiment 1.
Embodiment 4 preparations III procollagen type N end of the present invention peptide quantitative determination reagent kit
As outside the carrier, magnetic-particle adopts glutaraldehyde method bag quilt divided by magnetic-particle, and all the other all prepare III procollagen type N end peptide quantitative determination reagent kit with the method identical with embodiment 1.
The using method of embodiment 5 kits of the present invention
The concrete operations of the III procollagen type N end peptide quantitative determination reagent kit of above embodiment 1 preparation are as follows:
1) in 4 ℃ of refrigerators, takes out kit, equilibrium at room temperature 15 minutes.
2) take out coated slab, insert on the grillage.
3) add each concentration calibration object and testing sample in the reacting hole respectively, every hole adds sample and each standard items 0,6.2,12.5,25,50, each 25 μ l of 100ng/ml, blank 1 hole is established in each experiment, enzyme-added label 100 μ l in each hole except that blank well then, with the micro-oscillator mixing that fully vibrates, 37 ℃ of incubations 1 hour.
4) get rid of dereaction liquid, the cleansing solution after the dilution is filled it up with in every hole, washes plate 5 times, buckles on clean thieving paper at last and does.
5) each hole adds chemical luminous substrate liquid 50 μ l, and fully vibrating with micro-oscillator mixes, room temperature (20～25 ℃) lucifuge reaction 5 minutes.
6) add in behind the chemical luminous substrate liquid the 5th～30 minute and measure, on the chemiluminescence measuring instrument, measure the luminous intensity (RLU) in each hole in regular turn, 1 second/hole of Measuring Time.
7) be horizontal ordinate with calibration object concentration, the RLU value is drawn typical curve for ordinate, finds the concentration of this intraserous III procollagen type N end peptide on typical curve with each test serum RLU value.
The parallel contrast test of embodiment 6 kits of the present invention and existing market main flow reagent
(1) preparation of kit
A) preparation such as the embodiment 1 of III procollagen type N end peptide detection kit (chemoluminescence method);
B) choose that current market share is higher, the III procollagen type amino terminal peptide enzyme-linked immunologic detecting kit of main flow;
(2) detect sample
Normal control sample 180 examples are made a definite diagnosis oxyhepatitis sample 50 examples, chronic hepatitis sample 84 examples, and liver fibrosis sample 155 examples, diagnosis meets the liver fibrosis standard in the nineteen ninety-five Chinese Medical Association " virus hepatitis is prevented and treated scheme ".
(3) detection method
Adopt above-mentioned two kinds of III procollagen type amino terminal peptide detectable to carry out parallel detection, trace routine and result's judgement are carried out in strict accordance with each reagent instructions.
Above-mentioned two kinds of III procollagen type amino terminal peptide detection kit are carried out accuracy, specificity, sensitivity, accuracy performance index evaluations such as (recovery) respectively.
(4) testing result
The performance index evaluation of two kinds of III procollagen types of table 1 amino terminal peptide detectable
Two kinds of III procollagen types of table 2 amino terminal peptide detectable clinical sample content detection result (unit: ng/ml)
| Kit of the present invention | Enzyme-linked immunologic detecting kit | |
| Normal control sample (n=180) | 2.37±0.96 | 2.63±1.60 |
| Oxyhepatitis sample (n=50) | 2.88±1.76 | 3.59±2.19 |
| Chronic hepatitis sample (n=84) | 26.26±13.17 | 29.4±15.52 |
| Liver fibrosis sample (n=150) | 32.03±15.06 | 38.57±18.92 |
The testing result of two kinds of III procollagen types of table 3 amino terminal peptide detectable gathers
The described data of table 1-3 show that kit accuracy of the present invention, specificity, the recovery are better than market mainstream III procollagen type amino terminal peptide enzyme-linked immunologic detecting kit, and sensitivity significantly is better than market mainstream enzyme-linked immunologic detecting kit.Enzyme-linked immunologic detecting kit detects 2 parts of false positives, 3 parts of false negatives, and kit testing result of the present invention conforms to fully with clinical effectiveness, is 98.92% with the clinical coincidence rate of enzyme-linked immunologic detecting kit, related coefficient is R
2=0.9232 (see figure 2).
Claims (9)
1, a kind of III procollagen type N end peptide chemiluminescence immune analysis quantitative determination reagent kit is characterized in that described kit comprises:
1) III procollagen type N end peptide calibration object;
2) be coated with the solid phase carrier that III procollagen type N holds the monoclonal antibody of peptide;
3) monoclonal antibody of the III procollagen type N of enzyme labeling end peptide;
4) chemical luminous substrate that above-mentioned enzyme acted on; And
5) concentrated cleaning solution.
2, kit as claimed in claim 1 is characterized in that, described solid phase carrier is microwell plate, plastic bead, plastic tube or magnetic-particle.
3, kit as claimed in claim 1 is characterized in that, described enzyme is alkaline phosphatase or horseradish peroxidase; Described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, luminol or different luminol.
4, kit as claimed in claim 1 is characterized in that,
Described chemical luminous substrate liquid comprises 24g Tris, 160g NaCl, 4g KCl, 15mL HCl, 200mL 3-(2 '-spiral diamantane)-4-methoxyl-4-(3 " phosphorus acyloxy) phenyl-1,2-two oxidative ethanes (being AMPPD), 1mLProclin 300,1000mL distilled water;
Perhaps,
Described chemical luminous substrate comprises A liquid and B liquid, wherein,
Based on the described chemical luminous substrate A of 1000mL liquid, comprise 1.7716g luminol, 0.051g 4-xenol, 0.012g 4-iodobenzene boric acid, 11.4g boric acid, 4.9g borax, its pH value is 8.0～10.0;
Based on the described chemical luminous substrate B of 1000mL liquid, comprise 0.329g urea peroxide, 1ml Tween20,51.58g Na
2HPO
412H
2O, 8.74g NaH
2PO
42H
2O, its pH value is 7.0～7.6.
5, a kind of method for preparing the described kit of claim 1 is characterized in that may further comprise the steps:
1) with the pure product preparation of III procollagen type N end peptide III procollagen type N end peptide calibration object;
2) monoclonal antibody of holding peptide with enzyme labeling III procollagen type N;
3) hold the monoclonal antibody bag of peptide by solid phase carrier with III procollagen type N;
4) preparation chemical luminous substrate;
5) preparation concentrated cleaning solution;
6) monoclonal antibody of the III procollagen type N end peptide of the above-mentioned III procollagen type of packing N end peptide calibration object, enzyme labeling, chemical luminous substrate and the concentrated cleaning solution that this enzyme acted on; And
7) be assembled into finished product.
6, method as claimed in claim 5 is characterized in that, described bag is comprised following method by the step 3) of solid phase carrier:
1) bag quilt
The bag for preparing is cushioned liquid mixes, and the gained mixed liquor is carried on the solid phase carrier with the monoclonal antibody of III procollagen type N end peptide;
2) with physiological saline washing above-mentioned solid phase carrier; And
3) sealing
The confining liquid of preparation is sealed solid phase carrier after the above-mentioned washing.
As claim 5 or 6 described methods, it is characterized in that 7, described solid phase carrier is microwell plate, plastic bead, plastic tube or magnetic-particle.
8, method as claimed in claim 5 is characterized in that, described enzyme is alkaline phosphatase or horseradish peroxidase; Described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, luminol or different luminol.
9, method as claimed in claim 8, it is characterized in that described 1,2-two oxidative ethane analog derivatives are (diamantane)-1,2-two oxidative ethanes, 3-(2 '-the spiral diamantane)-4-methoxyl-4-(3 " the phosphorus acyloxy) phenyl-1,2-two oxidative ethanes, CSPD or CDP-Star.
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| CN 200810102020 CN101377509A (en) | 2008-03-14 | 2008-03-14 | III type precollagen N end peptide chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof |
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| CN105866415A (en) * | 2016-05-18 | 2016-08-17 | 北京北方生物技术研究所有限公司 | I-type collagen n-terminal peptide quantitative detection kit and preparation method thereof |
| CN106053826A (en) * | 2016-05-12 | 2016-10-26 | 威海威高生物科技有限公司 | III type procollagen N-terminal peptide quantitative measurement kit and preparation method thereof |
| CN106526165A (en) * | 2016-11-08 | 2017-03-22 | 北京久峰润达生物技术有限公司 | Quantitative measurement kit for glypican and detection method thereof |
| CN106855574A (en) * | 2016-06-30 | 2017-06-16 | 深圳市亚辉龙生物科技股份有限公司 | A kind of III procollagen type N-terminal peptide chemiluminescence immunity detection reagent and preparation method thereof |
| CN108431606A (en) * | 2016-02-03 | 2018-08-21 | 北欧生物科技公司 | The joint biomarker of fibrosis measures |
| CN109444430A (en) * | 2018-12-13 | 2019-03-08 | 郑州安图生物工程股份有限公司 | A kind of kit detecting III procollagen type N-terminal peptide |
| CN111004325A (en) * | 2020-01-02 | 2020-04-14 | 安徽合创健康生物技术有限公司 | III type procollagen N-terminal peptide nano antibody, and coding sequence and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108431606A (en) * | 2016-02-03 | 2018-08-21 | 北欧生物科技公司 | The joint biomarker of fibrosis measures |
| CN108431606B (en) * | 2016-02-03 | 2022-04-05 | 北欧生物科技公司 | Combined biomarker measurement of fibrosis |
| CN106053826A (en) * | 2016-05-12 | 2016-10-26 | 威海威高生物科技有限公司 | III type procollagen N-terminal peptide quantitative measurement kit and preparation method thereof |
| CN105866415A (en) * | 2016-05-18 | 2016-08-17 | 北京北方生物技术研究所有限公司 | I-type collagen n-terminal peptide quantitative detection kit and preparation method thereof |
| CN106855574A (en) * | 2016-06-30 | 2017-06-16 | 深圳市亚辉龙生物科技股份有限公司 | A kind of III procollagen type N-terminal peptide chemiluminescence immunity detection reagent and preparation method thereof |
| CN106526165A (en) * | 2016-11-08 | 2017-03-22 | 北京久峰润达生物技术有限公司 | Quantitative measurement kit for glypican and detection method thereof |
| CN111602056A (en) * | 2017-10-20 | 2020-08-28 | 北欧生物科技公司 | Collagen type XVI assay |
| CN111602056B (en) * | 2017-10-20 | 2024-04-23 | 北欧生物科技公司 | XVI collagen assay |
| CN109444430A (en) * | 2018-12-13 | 2019-03-08 | 郑州安图生物工程股份有限公司 | A kind of kit detecting III procollagen type N-terminal peptide |
| CN111004325A (en) * | 2020-01-02 | 2020-04-14 | 安徽合创健康生物技术有限公司 | III type procollagen N-terminal peptide nano antibody, and coding sequence and application thereof |
| CN111004325B (en) * | 2020-01-02 | 2021-07-06 | 安徽合创健康生物技术有限公司 | III type procollagen N-terminal peptide nano antibody, and coding sequence and application thereof |
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