CN201600371U - Beta2-microglobulin detecting kit - Google Patents
Beta2-microglobulin detecting kit Download PDFInfo
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- CN201600371U CN201600371U CN2009202170571U CN200920217057U CN201600371U CN 201600371 U CN201600371 U CN 201600371U CN 2009202170571 U CN2009202170571 U CN 2009202170571U CN 200920217057 U CN200920217057 U CN 200920217057U CN 201600371 U CN201600371 U CN 201600371U
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- nitrocellulose filter
- monoclonal antibody
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- beta2m
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Abstract
The utility model relates to the field of immunology detection, in particular to a beta2-microglobulin detecting kit consisting of water-absorbing filter paper (1), a nitrocellulose membrane (2), a colloidal gold pad (3), a sample pad (4) and a reaction supporting object (5); the nitrocellulose membrane (2) comprises a beta2M monoclonal antibody strip T, and a sheep anti-mouse IgG polyclonal antibody C; and the colloidal gold pad (3) contains a beta2M monoclonal antibody marked with colloidal gold. The beta2-microglobulin detecting kit can simplify beta2M detecting process. When being used for detecting beta2M, the kit does not have independence on any experimental apparatus, the environment or operators, is simple and convenient for operation, is short in detecting cycle, is easy to interpret, does not need special apparatus and equipment, does not need professional training, has strong adaptability, is convenient for monitoring and testing at anytime and anyplace.
Description
Technical field
The utility model relates to the immunology detection field, specially refers to a kind of B2M detection kit
Background technology
β2Wei Qiudanbai, hereinafter to be referred as β 2M, it is β 2 light chains in the HLA I class antigen, and it does not insert cell membrane and is free on the extracellular, and its function is relevant with the expression of stable I class antigen.Its gene is positioned on the 15th pair of chromosome, and the α heavy chain gene of HLA I class antigen then is positioned on the 6th pair of chromosome.β 2M expression of gene not exclusively is limited by the expression of I class antigen.The non-glycosylated single chain polypeptide that β 2M is made up of 100 amino acid residues, molecular mass 11800 μ.
The normal cell of human body interstitial cell, epithelial cell and hemopoietic system, malignant cell all can synthesize β 2M, and in vitro culture shows that lymphocyte receptor PHA-P HA stimulates the synthetic acceleration of back β 2M.β 2M level is regulated by α, gamma interferon mainly in the cell, and these media are synthetic transcriptional level control β 2M's.Liver is the major organs of synthetic β 2M, and the synthesis rate of normal person β 2M is very constant, about 0.13 (0.11～0.18) mg/ (hkg) body weight.Renal insufficiency patient synthesis rate is similar to the normal person.
β 2M is distributed widely in serum, cerebrospinal fluid, saliva, colostrum, amniotic fluid, seminal fluid and the urine in vivo.Child's serum content adult is slightly high, descends gradually with its serum content of age growth, and the normal adult blood level is very low, on average is about 1.5mg/L.
In the eubolism process, β 2M be discharged into body fluid after HLA separates, the β 2M 90% that goes into blood circulation exists with the non-protein combination state of monomer, its plasma half-life is less than 2h.β 2M is mainly from renal excretion, and 95% circulation β 2M can freely filter through glomerulus, is wherein absorbed with the pinocytosis form by the near-end renal tubule more than 99.9%, is transported to lysosomal enzyme after the absorption and is degraded to amino acid.Drain β 2M seldom, urine concentration＜0.2mg/L in normal person's urine.
When synthetic increasing or renal excretion when reducing in the β 2M body, can cause that blood β 2M increases.Numerous diseases such as malignant tumour, rheumatism, hepatopathy, chronic inflammation all can increase because of β 2M in the body is synthetic and cause that blood β 2M increases, and should at first get rid of these diseases when therefore using blood β 2M estimation renal function.
β 2M molecular mass is little, can freely pass through glomerular filtration, and only by renal excretion and decomposition, and produce constant airspeed in the body, be not subjected to how much influencing of age, sex, body musculature, therefore measure blood β 2M can reflect glomerular filtration rate(GFR (GFR) better than serum creatinine variation.Discover, the remarkable negative correlation of blood β 2M and GFR, and with serum creatinine, the remarkable positive correlation of urea nitrogen.Blood β 2M evaluate renal function is more responsive than serum creatinine.Blood β 2M measures can find earlier that the glomerulus function is impaired.When various former or Secondary cases glomerulus pathologies are involved the glomerular filtration function, blood β 2M is increased.
Blood β 2M reflects that the slight renal function of diabetic goes down and the sensitive indicator of observe the curative effect, and blood β 2M can reflect the impaired renal function situation of high blood pressure in early days, to the clinical guidance medication, it is significant to avoid using the depressor and the microbiotic that influence renal function.CH patient blood β 2M significantly increases, and high-caliber blood β 2M is relevant with the generation of amyloidosis, amyloid osteoarthropathy and carpal tunnel syndrome.
In 2,3 days, blood β 2M begins to raise, and descends gradually subsequently after the kidney transplant, and its decline rate is faster than serum creatinine.When rejection took place, blood β 2M rose suddenly once more.Therefore blood β 2M helps to dynamic observe and diagnoses kidney transplantation exclusion reaction in early days.
β 2M is synthetic in body increases, and blood β 2M level is higher than normal value more than 3 times the time, and the glomerular filtration amount surpasses the heavy receptivity of renal tubule, and then β 2M discharge is increased in the urine.Therefore, during with urine β 2M evaluation of measuring renal function, should survey blood β 2M, eliminating causes the disease that blood β 2M increases.If in the blood β 2M content normal and in the urine β 2M content increase, illustrate that kidney proximal tubule function is impaired.Discover that it is because the cause that kidney proximal tubule BBM albumen capture process is disturbed or lysosomal protein kalabolism reduces that urine β 2M increases.Measure urine β 2M and in the diagnosis and treatment of kidney trouble, have important value.
Urine β 2M is the index of the heavy absorption function of reflection renal tubule: the near-end renal tubule is unique place that β 2M handles in vivo, and slight when impaired when the near-end renal tubule, urine β 2M obviously increases, and urine β 2M and renal tubule weigh absorptivity and be proportionate.Therefore, measuring urine β 2M is impaired sensitivity, the special method of diagnosis near-end renal tubule.The cause of disease that causes the heavy malabsorption of near-end renal tubule is a lot, comprises that congenital renal tubule pathology and posteriority renal tubular function are impaired.
But the tubular injury of urine β 2M early detection diabetes and patients with hypertension: diabetes, patients with hypertension are urinated β 2M in early days and are promptly increased, and illustrate that diabetes and patients with hypertension promptly have tubular injury in early days.Urine β 2M increases and kidney function damage degree significant correlation, and the treatment back obviously descends with state of an illness control urine β 2M.
Urine β 2M can dynamic observe and diagnose early stage kidney transplantation exclusion reaction, if urine β 2M still raises behind the antirejection therapy, then often showing has rejection repeatedly, finally can cause the transplanted kidney function forfeiture.
The method of existing in the market detection β 2M is a chemoluminescence method, and the effect of this detection method relies on experimental apparatus, environment or operating personnel's proficiency, is not easy to monitor anywhere or anytime and check.
The utility model content
Technical problem to be solved in the utility model is to overcome the deficiencies in the prior art, provides a kind of simple to operation, laboratory condition is not had dependent, is used for the detection kit of the β 2M of fast detecting blood and urine.
For solving the problems of the technologies described above, the utility model is realized as follows: B2M detection kit described in the utility model is by absorbent filter, nitrocellulose filter, and the collaurum pad, sample pad, the reaction holder is formed; Described nitrocellulose filter comprises β 2M monoclonal antibody band T and sheep anti-mouse igg polyclonal antibody C; Described collaurum pad contains the β 2M monoclonal antibody of colloid gold label; Bag is sticked on the middle part of reaction holder by good nitrocellulose filter; Be pasted with absorbent filter on the reaction holder of band C line direction, push down the nitrocellulose filter certain width; At first paste on the reaction holder of band T line direction proper width the collaurum pad, push down the edge certain width of nitrocellulose filter band T line; Sample pad is pasted in continuation in that this side up, pushes down collaurum pad proper width.
Described nitrocellulose filter can comprise β 2M monoclonal antibody band T and sheep anti-mouse igg polyclonal antibody C.
Described nitrocellulose filter can comprise β 2M polyclonal antibody band T and sheep anti-mouse igg polyclonal antibody C
Described collaurum pad can contain the β 2M monoclonal antibody of colloid gold label.
Described collaurum pad can contain the β 2M polyclonal antibody of colloid gold label.
Good effect of the present utility model is: B2M detection kit described in the utility model, can simplify β 2M testing process.Utilize this kit to detect β 2M, any experimental apparatus, environment or operating personnel are not had dependence, interpretation is lacked, is easy to simple, convenient, sense cycle, do not need special instrument and equipment, do not need professional training, adaptability is strong, is convenient to monitor anywhere or anytime and check.
Description of drawings
Below in conjunction with the drawings and specific embodiments the utility model is described in further detail.
Fig. 1 is the front schematic view of the utility model detection kit;
Fig. 2 is the side schematic view of the utility model detection kit;
Fig. 3 is the positive synoptic diagram of the utility model testing result;
Fig. 4 is the negative synoptic diagram of the utility model testing result;
Fig. 5 is the invalid synoptic diagram of the utility model testing result;
Fig. 6 is the invalid synoptic diagram of the utility model testing result;
Among the figure: 1 absorbent filter, 2 nitrocellulose filters, 3 collaurum pads
4 sample pad, 5 reaction holders
Embodiment
See figures.1.and.2, B2M detection kit described in the utility model is by absorbent filter 1, nitrocellulose filter 2, and collaurum pad 3, sample pad 4, reaction holder 5 is formed; Described nitrocellulose filter 2 comprises 2M monoclonal antibody band T and sheep anti-mouse igg polyclonal antibody C; Described collaurum pad 3 contains the β 2M monoclonal antibody of colloid gold label; Bag is sticked on the reaction holder 5 by good nitrocellulose filter 2, the Edge Distance reaction holder 5 edge 28.0mm of its band T line direction nitrocellulose filter, the edge 32.0mm of the Edge Distance reaction holder 5 of band C line direction nitrocellulose filter; Be pasted with absorbent filter 1 on the reaction holder 5 of band C line direction, wide is 33.0mm, pushes down nitrocellulose filter 1.0mm; At first paste the wide collaurum pad 3 of 6.0mm on the reaction holder 5 of band T line direction, push down the edge 1.0mm of nitrocellulose filter band T line; Sample pad 4 is pasted in continuation in that this side up, and wide is 25.0mm, pushes down collaurum pad 2.0mm.
At first prepare the B2M detection kit, the purifying of 2M monoclonal antibody ascites:
After will containing the sad-saturated ammonium sulphate of ascites use of β 2M monoclonal antibody, use ultraviolet spectrometry to measure protein concentration and SDS-PAGE mensuration purity, the protein concentration of gained monoclonal antibody is higher than 2.0mg/ml, and purity is qualified greater than 90%.The preparation of collaurum and with evaluation: use trisodium citrate reduction method reduction gold chloride to prepare colloid gold particle, use the absorbance value in ultraviolet spectrophotometer scanning 460～600nm interval, in 520 to 525nm intervals maximum absorption band is arranged, and absorption value is qualified greater than 2.0, and corresponding colloid gold particle diameter is approximately 30～40nm.Determining of β 2M monoclonal antibody optimum mark pH value and optimum mark concentration: use this antibody of the parallel mark of collaurum gradient pH value, select the variable color edge flag condition this pH value is added that 0.5 carries out the optimal pH of mark as this antibody, be approximately 7.4 to 8.0; Under this optimal pH condition, set the albumen gradient and carry out mark, mixing after the adding monoclonal antibody in collaurum, place more than 20 minutes, the ratio that adds 100 μ l in every milliliter of collaurum adds 10% NaCl, and mixing is observed color, the selected labelled amount that adds additionally additional again 10% value conduct the best of labelled amount that monoclonal antibody concentration is minimum and color does not change is approximately 12 to 18 μ g/ml.Colloid gold label purifying antibody: the colloidal gold solution that mark is good, add BSA and PEG20000 and be 0.2% to final concentration, mixing, place more than 15 minutes, use high speed freezing centrifuge 12000rpm centrifugal more than 15 minutes, supernatant discarded will precipitate and use collaurum to preserve the liquid redissolution, and volume is 1/10 of a former mark collaurum volume.The debugging of redissolution ratio: use collaurum working fluid dilution mark collaurum concentrate to dilute, dilution ratio is 10 to 20 times, spray to and carry out drying on the collaurum pad, last according to being determined best dilution ratio by the testing result that nitrocellulose filter matches with bag, generally select about 15 times.Coated antibody is best to be wrapped by the selection of condition: the T line on the nitrocellulose filter and the bag of C line are provided with several concentration gradients by concentration and wrap quilt respectively, the final best conduct the best of testing result of selecting is wrapped by concentration, and bag is debugged with damping fluid, package amount etc.It is 0.02M pH 7.2PBS that final selection bag is cushioned liquid, and bag is 1.0mg/ml by concentration, and discharge rate is chosen as 1.0 μ l/cm.
Sample to be checked (whole blood, serum, blood plasma or urine) 60 to 100 μ l are directly added detection kit sample pad 4 places (extra some dilutions that add of whole blood sample needs), sample will move to the absorbent filter direction along each attachment of reaction holder 5, reads experimental result in 20 minutes.
With reference to Fig. 3, if contain β 2M in the sample, then combine and form compound with the β 2M monoclonal antibody specificity of colloid gold label on the detection kit, along the film district continue up then be coated on nitrocellulose filter 2 on another β 2M monoclonal antibody generation specificity combine, occur red or pink at band T place
With reference to Fig. 4, if there is not β 2M antigen in the sample, just can not form above-mentioned compound, band T place also just can not outlet redness or pink, negative.
With reference to Fig. 5 and Fig. 6, no matter whether contain β 2M in the sample, the monoclonal antibody of colloid gold label all can continue to go upward to the band C of coated film, remove to wrap the sheep anti-mouse igg of quilt herein in conjunction with forming red or pink band, be nature controlling line,, prove that then collaurum lost efficacy or operation makes mistakes if this band does not occur in detection, the result is invalid, needs to detect again.
The specific embodiment of the above is intended to specify thinking of the present utility model, and the enforcement of the utility model is not limited to the disclosed mode of above specific embodiment.All mentalities of designing based on the utility model are simply deduced and are replaced, and all belong to the enforcement of the utility model.
Claims (3)
1. B2M detection kit is characterized in that: it is by absorbent filter (1), nitrocellulose filter (2), and collaurum pad (3), sample pad (4), reaction holder (5) is formed; Described nitrocellulose filter (2) comprises β 2M monoclonal antibody band T and sheep anti-mouse igg polyclonal antibody C; Described collaurum pad (3) contains the β 2M monoclonal antibody of colloid gold label; Bag is sticked on the reaction holder (5) by good nitrocellulose filter (2), Edge Distance reaction holder (5) the edge 28.0mm of its band T line direction nitrocellulose filter, the edge 32.0mm of the Edge Distance reaction holder (5) of band C line direction nitrocellulose filter; Be pasted with absorbent filter (1) on the reaction holder (5) of band C line direction, push down nitrocellulose filter 1mm to 2mm; At first paste the collaurum pad (3) of 4mm to 8mm on the reaction holder (5) of band T line direction, the edge 1mm that pushes down nitrocellulose filter band T line is to 2mm; Sample pad (4) is pasted in continuation in that this side up, pushes down collaurum pad 2mm to 4mm.
2. B2M according to claim 1 (β 2M) detection kit is characterized in that: described nitrocellulose filter (2) comprises β 2M polyclonal antibody or monoclonal antibody band T and sheep anti-mouse igg polyclonal antibody C.
3. B2M detection kit according to claim 1 is characterized in that: described collaurum pad (3) contains the β 2M monoclonal antibody or the polyclonal antibody of colloid gold label.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009202170571U CN201600371U (en) | 2009-09-25 | 2009-09-25 | Beta2-microglobulin detecting kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009202170571U CN201600371U (en) | 2009-09-25 | 2009-09-25 | Beta2-microglobulin detecting kit |
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| Publication Number | Publication Date |
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| CN201600371U true CN201600371U (en) | 2010-10-06 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN2009202170571U Expired - Lifetime CN201600371U (en) | 2009-09-25 | 2009-09-25 | Beta2-microglobulin detecting kit |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102590526A (en) * | 2012-01-13 | 2012-07-18 | 宁波美康生物科技股份有限公司 | Beta 2-microglobulin detection kit |
| CN103509760A (en) * | 2013-10-10 | 2014-01-15 | 深圳市菲鹏生物股份有限公司 | Hybridoma cell capable of secreting anti-beta2-microglobulin monoclonal antibody, and application thereof |
| CN106443012A (en) * | 2016-09-12 | 2017-02-22 | 三诺生物传感股份有限公司 | Test strip, preparation method thereof and application of test strip to combined detection of microalbuminuria and beta2 microglobulin |
-
2009
- 2009-09-25 CN CN2009202170571U patent/CN201600371U/en not_active Expired - Lifetime
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102590526A (en) * | 2012-01-13 | 2012-07-18 | 宁波美康生物科技股份有限公司 | Beta 2-microglobulin detection kit |
| CN102590526B (en) * | 2012-01-13 | 2014-03-12 | 宁波美康生物科技股份有限公司 | Beta 2-microglobulin detection kit |
| CN103509760A (en) * | 2013-10-10 | 2014-01-15 | 深圳市菲鹏生物股份有限公司 | Hybridoma cell capable of secreting anti-beta2-microglobulin monoclonal antibody, and application thereof |
| CN103509760B (en) * | 2013-10-10 | 2015-04-22 | 深圳市菲鹏生物股份有限公司 | Hybridoma cell capable of secreting anti-beta2-microglobulin monoclonal antibody, and application thereof |
| CN106443012A (en) * | 2016-09-12 | 2017-02-22 | 三诺生物传感股份有限公司 | Test strip, preparation method thereof and application of test strip to combined detection of microalbuminuria and beta2 microglobulin |
| CN106443012B (en) * | 2016-09-12 | 2018-11-06 | 三诺生物传感股份有限公司 | A kind of test strips and preparation method thereof and the application in microdose urine protein and β2-microglobulin joint-detection |
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| C14 | Grant of patent or utility model | ||
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| DD01 | Delivery of document by public notice |
Addressee: Gu Ziyi Document name: Notification to Pay the Fees |
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| DD01 | Delivery of document by public notice |
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| DD01 | Delivery of document by public notice |
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| ASS | Succession or assignment of patent right |
Owner name: BEIJING EASYSWEET BIOMEDICINE SCITECH CO., LTD. Free format text: FORMER OWNER: GU ZIYI Effective date: 20150603 |
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| C41 | Transfer of patent application or patent right or utility model | ||
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Free format text: CORRECT: ADDRESS; FROM: 034100 XINZHOU, SHAANXI PROVINCE TO: 102629 DAXING, BEIJING |
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Effective date of registration: 20150603 Address after: 102629 Beijing City, Daxing District science and Technology Park of Zhongguancun Daxing biomedical industry base of Yongxing Road No. 25 Patentee after: Beijing Easysweet Biomedicine SciTech Co., Ltd. Address before: 034100 No. 189 sports North Road, Shanxi, Yuanping Patentee before: Gu Ziyi |
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| CP01 | Change in the name or title of a patent holder |
Address after: 102629 Beijing City, Daxing District science and Technology Park of Zhongguancun Daxing biomedical industry base of Yongxing Road No. 25 Patentee after: Beijing Easysweet Biotechnology Co., Ltd. Address before: 102629 Beijing City, Daxing District science and Technology Park of Zhongguancun Daxing biomedical industry base of Yongxing Road No. 25 Patentee before: Beijing Easysweet Biomedicine SciTech Co., Ltd. |
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Granted publication date: 20101006 |