CN110389233A - It is a kind of for detecting the FN class antibody mediated immunity detection kit of people AMH - Google Patents
It is a kind of for detecting the FN class antibody mediated immunity detection kit of people AMH Download PDFInfo
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- CN110389233A CN110389233A CN201810349605.XA CN201810349605A CN110389233A CN 110389233 A CN110389233 A CN 110389233A CN 201810349605 A CN201810349605 A CN 201810349605A CN 110389233 A CN110389233 A CN 110389233A
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- 230000015572 biosynthetic process Effects 0.000 claims 1
- 238000003745 diagnosis Methods 0.000 abstract description 10
- 239000012472 biological sample Substances 0.000 abstract description 2
- 101001011645 Homo sapiens Muellerian-inhibiting factor Proteins 0.000 description 6
- 206010033266 Ovarian Hyperstimulation Syndrome Diseases 0.000 description 4
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- 101150070496 Amh gene Proteins 0.000 description 2
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- 239000000868 anti-mullerian hormone Substances 0.000 description 1
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Endocrinology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to a kind of FN class antibody mediated immunity detection kits for detecting people AMH.Kit includes anti-Miao Le Shi pipe hormone antibody, the standard items, calibration object, phosphate solution of the coated magnetic particle of Miao's Le Shi pipe hormone antibody, alkali phosphatase enzyme mark.This kit can be used to detect the level of the AMH in people's biological sample, and may be used as diagnosis/prognostic indicator.
Description
Technical field
The invention belongs to biomedicine technical fields, are related to a kind of detection examination for detecting the anti-Miao Le Shi pipe hormone (AMH) of people
Agent box may be used as the prediction in Controlled ovarian super-stimulation (COH) effect, the prediction of ovarian hyperstimulation (OHSS), polycystic ovary
The diagnosis and prognosis of syndrome (PCOS), the diagnosis of granulosa cell tumor of ovary and management, male precocity and delayed puberty
The antidiastole of the double character states of diagnosis and differential diagnosis, the diagnosis of the cryptorchidism of boy and anorchia and children.
Background technique
With the change of modern way of life, universal late marriage and childbirth phenomenon, undesirable modern way of life are (as excessively saved
Food weight-reducing, Smoking And Drinking, stay up late), huge work ethos pressure, with comprehensive relieving of national child-bearing policy, China is above the average age for marriage
Puerpera's number increases substantially, and hard to settle, infertile crowd's specific gravity increase, puzzlement of more and more women by fecundity problem.
Clinically domestic each diagnosis and treatment unit assessment ovarian function mainly includes age, antral follicle count (AFC) and basic sex hormone at present
Horizontal (FSH, FSH/LH, E2, inhibin B etc.).But these indexs all have some limitations: AFC will receive B ultrasound operation
The influence of the subjective factor of person and different female pelvic cavity environment, leads to the deviation of ultrasound image result.Basic sex hormone water
The variation of flat (including FSH, FSH/LH ratio, E2 and inhibin B) is influenced by the menstrual cycle, and variation occurs later.
In comparison, in recent years in American-European countries, clinically gradually begin to use anti-Miao Leguan hormone (Anti-Mullerian
Hormone:AMH) the Xiang Zhibiao as ovarian function assessment.
Anti- gyneduct hormone abbreviation AMH, is one of the member of β-transforming factor (TGF-β) family, by Profess
Alfred Jost is in nineteen forty-seven first discovery, also referred to as gyneduct inhibiting factor (MIF), gyneduct inhibitory hormone
(MIH) and gyneduct mortifier (MIS).AMH albumen is encoded by AMH gene, and AMH gene is a kind of positioned at No. 19 chromosomes
The mini gene of 2.4-2.8Kb on galianconism has 5 exons.AMH is to be joined by 2 70kDa monomers by disulfide bond before secretion
Connect the dimer to form a 140kDa.During passing through cell membrane, one specific site of AMH precursor is converted by proteinogen
Enzyme cuts into N-terminal proparea and C-terminal maturation zone, and together, C-terminal is active function end to this two parts still Non-covalent binding.
Fibronectin (Fibronectin, FN), with 3 similar complementary determining regions of conventional monoclonal antibody
(complementarity determining region, CDR), the i.e. combined area of antibody and antigen, therefore FN class antibody has
Specific antigen associativity as having antibody class.But its molecular weight is only 10Kd, is 1/the 15 of conventional monoclonal antibody, and nothing
Disulfide bond and glycosylation modified has stronger structural stability, and it is expensive to can be used cheap escherichia expression system to substitute
Animal cell expression, greatly reduce production cost.Additionally due to the area FC of antibody is not contained, not by complement in detection process
Interference.FN class antibody has huge advantage in exploitation immunologic function test reagent.
Summary of the invention
The present invention relates to a kind of FN class antibody mediated immunity detection kit for detecting people AMH, may be used as in the super row of controlling
The prediction of ovum (COH) effect, the prediction of ovarian hyperstimulation (OHSS), the diagnosis of Stein-Leventhal syndrome (PCOS) and prognosis,
The diagnosis and management of granulosa cell tumor of ovary, the diagnosis and differential diagnosis of male precocity and delayed puberty, boy cryptorchidism
The antidiastole of the diagnosis of disease and anorchia and the double character states of children.
The present invention provides the FN class antibody and recombined human AMH egg of AMH in immunochemiluminescence method method detection biological sample
White, of the invention kit further include: anti-Miao Le Shi of the coated magnetic particle of Miao's Le Shi pipe hormone antibody, alkali phosphatase enzyme mark
Pipe hormone antibody, standard items, calibration object, phosphate solution.Further, the FN class antibody of anti-human AMH provided by the invention is to adopt
It is obtained with escherichia expression system.
The development process of mentioned reagent box provided by the invention is as follows:
(a) preparation of recombined human AMH albumen
Recombined human AMH design of primers → recombined human AMH strain construction → colibacillus high expression expression system foundation → parent
With ion chromatography → obtain 95% recombined human AMH albumen of purity >
(b) preparation of recombined human AMH detection kit antibody
The external display technique of mRNA establishes anti-human AMH antibody library → screening high specific, high-affinity antibody segment → structure
Build bacterial strain → FN antibody affinity, the specificity, physicochemical property (fusing point, polymerism) detection → large intestine of expression FN candidate antibodies
Bacillus expression system prepares the amplification research of FN antibody.
Specific embodiment
Embodiment 1: kit of the invention can be following substance:
(1) Miao Le Shi pipe hormone antibody coated magnetic particle, contains preservative by 3.5mL/ bottles；
(2) anti-Miao Le Shi pipe hormone antibody alkali phosphatase enzyme mark conjugate, contains preservative by 4.0mL/ bottles；
(3) calibration object, 1 bottle；
(4) phosphate solution contains protein stabiliser；
(5) containing anti-Miao Le Shi pipe neurophysin, protein stabiliser, preservative dried frozen aquatic products；
Embodiment 2: sample process
Serum processing: room temperature blood natural coagulation 10 minutes, 2000 revs/min are centrifuged 20 minutes or so.Supernatant is carefully collected,
As precipitated during preservation, should be centrifuged again.
Urine process: being collected with sterile tube, and 3000 revs/min are centrifuged 20 minutes or so.Supernatant is carefully collected, during preservation
It is formed, should be centrifuged again if any precipitating.
Tissue specimen processing: after cutting sample, weight is weighed.PH7.4PBS is added in volume ratio 1:1 by weight, fast with liquid nitrogen
Quickly cooling jelly saves backup.Sample still maintains 2-8 DEG C of temperature after melting.Be added same volume PBS (pH7.4), by hand or
Sample is homogenized abundant by homogenizer.3000 revs/min are centrifuged 20 minutes or so, carefully collect supernatant.It is a to be detected after packing,
Waste cold is frozen spare.It is extracted as early as possible after collection of specimens, extracts and carried out by pertinent literature, should be tested as early as possible after extraction.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all in spirit of the invention and
Within principle, any modification, equivalent replacement, improvement and so on should be included in protection scope of the present invention.
Claims (4)
1. a kind of FN class antibody mediated immunity detection kit for detecting people AMH, consisting of:
The coated magnetic particle of anti-Miao Le Shi pipe hormone antibody, anti-Miao Le Shi pipe hormone antibody of alkali phosphatase enzyme mark, standard items,
Calibration object, phosphate solution；
Wherein the antibody formation in the coated magnetic particle of anti-Miao Le Shi pipe hormone antibody is FN class antibody.
Wherein anti-Miao Le Shi pipe hormone antibody form of the alkali phosphatase enzyme mark is FN class antibody.
2. according to detection kit described in right 1, it is characterised in that:
(1) AMH is anti-Miao Le Shi pipe hormone, is one of the member of β-transforming factor (TGF-β) family；
(2) FN class antibody is fibronectin (Fibronectin, FN).
3. FN class antibody according to claim 2, it is characterised in that molecular weight is only 10Kd, is the 15 of conventional monoclonal antibody
/ mono-, and without disulfide bond and glycosylation modified.
4. kit, application method described in claim any one of 1-3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810349605.XA CN110389233A (en) | 2018-04-18 | 2018-04-18 | It is a kind of for detecting the FN class antibody mediated immunity detection kit of people AMH |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810349605.XA CN110389233A (en) | 2018-04-18 | 2018-04-18 | It is a kind of for detecting the FN class antibody mediated immunity detection kit of people AMH |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110389233A true CN110389233A (en) | 2019-10-29 |
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|---|---|---|---|
| CN201810349605.XA Pending CN110389233A (en) | 2018-04-18 | 2018-04-18 | It is a kind of for detecting the FN class antibody mediated immunity detection kit of people AMH |
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| Country | Link |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110027235A1 (en) * | 2009-04-09 | 2011-02-03 | Sangamo Biosciences, Inc. | Targeted integration into stem cells |
| CN103703370A (en) * | 2011-05-27 | 2014-04-02 | 北欧生物科技公司 | Detection of diagnostic peptides |
| CN105891490A (en) * | 2016-04-05 | 2016-08-24 | 付国亮 | Test strip for quantitatively detecting anti-mullerian hormone, preparation method thereof and determination method for concentration of anti-mullerian hormone |
| CN106053791A (en) * | 2016-06-30 | 2016-10-26 | 深圳市亚辉龙生物科技股份有限公司 | Anti-mullerian hormone chemiluminescence immunoassay kit and preparation method and application thereof |
-
2018
- 2018-04-18 CN CN201810349605.XA patent/CN110389233A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110027235A1 (en) * | 2009-04-09 | 2011-02-03 | Sangamo Biosciences, Inc. | Targeted integration into stem cells |
| CN103703370A (en) * | 2011-05-27 | 2014-04-02 | 北欧生物科技公司 | Detection of diagnostic peptides |
| CN105891490A (en) * | 2016-04-05 | 2016-08-24 | 付国亮 | Test strip for quantitatively detecting anti-mullerian hormone, preparation method thereof and determination method for concentration of anti-mullerian hormone |
| CN106053791A (en) * | 2016-06-30 | 2016-10-26 | 深圳市亚辉龙生物科技股份有限公司 | Anti-mullerian hormone chemiluminescence immunoassay kit and preparation method and application thereof |
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