WO2016101847A1 - 一种慢性肾病诊断试剂盒 - Google Patents

一种慢性肾病诊断试剂盒 Download PDF

Info

Publication number
WO2016101847A1
WO2016101847A1 PCT/CN2015/097926 CN2015097926W WO2016101847A1 WO 2016101847 A1 WO2016101847 A1 WO 2016101847A1 CN 2015097926 W CN2015097926 W CN 2015097926W WO 2016101847 A1 WO2016101847 A1 WO 2016101847A1
Authority
WO
WIPO (PCT)
Prior art keywords
fgf23
kit
detection
biontin
nhs
Prior art date
Application number
PCT/CN2015/097926
Other languages
English (en)
French (fr)
Inventor
李校堃
梁广
王晓杰
李世军
Original Assignee
温州医科大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 温州医科大学 filed Critical 温州医科大学
Priority to US15/300,236 priority Critical patent/US20170184593A1/en
Publication of WO2016101847A1 publication Critical patent/WO2016101847A1/zh

Links

Images

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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6872—Intracellular protein regulatory factors and their receptors, e.g. including ion channels
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/475—Assays involving growth factors
    • G01N2333/50—Fibroblast growth factors [FGF]
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/34—Genitourinary disorders
    • G01N2800/347—Renal failures; Glomerular diseases; Tubulointerstitial diseases, e.g. nephritic syndrome, glomerulonephritis; Renovascular diseases, e.g. renal artery occlusion, nephropathy

Definitions

  • the invention belongs to the technical field of disease detection, and particularly relates to a diagnostic kit for chronic kidney disease.
  • Fibroblast growth factor 23 (FGF23), a member of the FGFs superfamily, plays an important role in the maintenance of early phosphorus homeostasis in patients with chronic kidney disease by inhibiting phosphorus reabsorption and increasing exocrine phosphorus secretion (J Am Soc Nephrol) .2007:18:1637-1647).
  • FGF23 Fibroblast growth factor 23
  • the American Journal of Kidney and New England have reported that elevated levels of FGF23 in patients with chronic kidney disease precede lung abnormalities; FGF23 expression levels are independently associated with worsening of chronic kidney disease and mortality in patients undergoing advanced dialysis. Therefore, FGF23 has become an important marker for early diagnosis of renal function.
  • the invention aims to provide a diagnostic kit for chronic kidney disease, and provides a simple and convenient method for early diagnosis of chronic kidney disease, and provides a new method and a path for detecting and treating chronic kidney disease.
  • a diagnostic kit for chronic kidney disease including FGF23 monoclonal antibody, biotin standard Reagents Sulfo-NHS-LC-Biontin, buffer solution, casein salt solution, streptomycin avidin and substrate TMB.
  • the kit of the present invention used pH 7.0 0.1 M PBS as a coating buffer solution, and the sample had a coating volume of 100 ⁇ l and a coating concentration of 2 ⁇ g/ml.
  • the FGF23 antibody of the present invention and the biotin labeling reagent Sulfo-NHS-LC-Biontin were labeled at a weight ratio of 1:5 to 1:12, and the concentration after mixing was 2 ⁇ g/ml.
  • the casein salt solution described in the kit of the present invention is a dilution of FGF23, a detection antibody and streptomycin, and the dilution factor of streptomycin is 1:3000.
  • the streptomycin avidin described in the kit of the present invention has a working time of 10 minutes with the substrate.
  • the minimum detection limit of the kit FGF23 of the present invention is 1 opg/ml, and the detection linear range is 2400 pg/ml to 1 opg/ml.
  • the invention has the beneficial effects of providing an in vitro detection kit for early detection of chronic kidney disease, the sensitivity of the kit to clinical detection of FGF23 is 1 opg/ml, which is 10 times higher than that of conventional monoclonal antibody, and for chronic kidney disease. Early diagnosis is of great significance.
  • Figure 1 is a linear relationship between absorbance values and sample concentrations in an embodiment of the present invention.
  • mice were immunized with FGF23-Fc antigen to obtain hybridoma cell-positive FGF23 antibody clones. 1276 clones were screened and their binding specificity to FGF23 protein was detected. Three types of 278, 6B12 and 6H1 were screened. antibody.
  • FGF23 monoclonal antibodies 278, 6B12 and 6H1 were screened for the optimization of the detection conditions of the FGF23 ELISA kit.
  • biomarker conditions were optimized around the optimal biotinylation conditions, 6B12 was biotinylated at 1:8, 1:6, 1:5, 1:4; 6H1 was 1:12, 1: 10, 1:8 for marking.
  • biomarker conditions were optimized around the optimal biotinylation conditions, 6B12 was biotinylated at 1:7, 1:8, 1:9; 6H1 was 1:9, 1:10, 1: 11 mark.
  • the data in Table 3 shows that the optimal ratio of antibody 6B12 for biotin labeling is 1:8; the optimal concentration of 6H1 for biotinylation labeling is 1:10 and 1:9.
  • the labeling ratio refinement was performed around the 6B12 and 6H1 optimal sensitivity label data, and 6B12 was biotinized at 1:8, 1:9, 1:10, 1:11, 1:12, 1:20.
  • the reaction was carried out; 6H1 was labeled 1:3, 1:4, 1:5, 1:6, 1:7, 1:10, and the experimental data is shown in Table 6.
  • the labeling ratio refinement was performed around the 6B12 and 6H1 optimal sensitivity label data, and 6B12 and 6H1 were biotinylated at 1:8, 1:9, 1:10, 1:11, 1:12, respectively.
  • Reaction, experimental data are shown in Table 7.
  • the experimental data in Table 7 shows that 6B12 selects 1:8 for labeling, and 6H1 selects 1:10 for labeling, the detection sensitivity is the highest.
  • the detection sensitivity and background level of the coating were investigated by using 0.1 M, pH 7.0 phosphate buffer solution, and pH 9.0, 0.1 M sodium borohydride solution as the coating solution.
  • the experimental data are shown in Table 9.
  • the experimental data of Table 9 shows that pH 7.0 phosphate buffer solution was selected as the coating solution of the FGF23 kit, taking into account the detection sensitivity and background level.
  • the kit biotin labeling reagent was selected as Sulfo-NHS-LC-Biontin (Lot #21327), the labeling condition was 6B12 selection 1:8 for labeling, 6H1 selection was 1:10 for labeling, biotin labeling antibody 6B12 and 6H1 were mixed and the concentration was 2 ⁇ g/ml.
  • the coating buffer solution Using pH 7.0, 0.1M PBS as the coating buffer solution, the coating volume of anti-FGF23 was 100 ⁇ l, and the coating concentration of anti-FGF23 was 2 ⁇ g/ml.
  • the sample dilution is selected casein salt solution, the dilution factor of streptomycin avidin is 1:3000, and the interaction time of the enzyme and the substrate is 10 minutes;
  • FGF23 was subjected to 3-fold gradient dilution at 10000 pg/ml and 2400 g/ml, and whether the p-value between the lowest detection concentration and the blank control well was ⁇ 0.05 was taken as the lowest detection limit of FGF23.
  • the concentration of the FGF23 standard was started at 2400 pg/ml, and subjected to three-fold gradient dilution.
  • the coating was carried out according to the above optimized conditions, and TMB was used for color development to study the linear relationship between the absorbance value and the sample concentration.
  • the experimental results are shown in Fig. 1.
  • the experimental data show that the linear range of the FGF23 detection kit is 2400 pg/ml to 10 pg/ml.

Abstract

一种慢性肾病诊断试剂盒，包括FGF23单抗隆抗体、生物素标记试剂Sulfo-NHS-LC-Biontin、pH 7.0 0.1M的PBS缓冲溶液、casein盐溶液、链霉素亲和素和底物TMB，FGF23抗体与生物素标记试剂Sulfo-NHS-LC-Biontin标记条件为重量比为1∶5～1∶12，链霉素亲和素的稀释倍数是1∶3000。试剂盒FGF23的最低检测限值为10pg/ml，比常规单抗提高了10倍的灵敏度，对于慢性肾病的早期诊断具有重要意义。

Description

一种慢性肾病诊断试剂盒 技术领域
本发明属于疾病检测技术领域，具体涉及一种慢性肾病诊断试剂盒。
背景技术
成纤维细胞生长因子23(FGF23)是FGFs超家族中的一员，它通过抑制磷的重吸收及增加磷的外分泌，在慢性肾病病人早期磷稳态的维持中发挥重要作用(J Am Soc Nephrol.2007：18：1637-1647)。《美国肾脏杂志》及《新英格兰》相继报道，慢性肾病病人FGF23浓度升高先于磷代谢异常；FGF23表达水平与慢性肾病的恶化及晚期透析治疗病人的死亡率成独立相关。因此，FGF23成为了肾功能早期诊断的重要标志物。由于目前临床使用的生物标物-肌氨酸酐存在着准确性低、特异性不高的缺陷，而且一旦查出肌氨酸酐水平超标，多数病程已经达到中晚期。因此，开发新的慢性肾病诊断试剂-FGF23单克隆抗体试剂盒具有重要的临床应用价值。
发明内容
本发明目的在于提供一种慢性肾病诊断试剂盒，为慢性肾病早期诊断提供一种简便易行的方法，为慢性肾病的检测和治疗提供了新方法与途径。
本发明具体通过以下技术方案实现：
一种慢性肾病诊断试剂盒，包括FGF23单抗隆抗体、生物素标 记试剂Sulfo-NHS-LC-Biontin、缓冲溶液、casein盐溶液、链霉素亲和素和底物TMB。
本发明试剂盒使用pH 7.0 0.1M PBS作为包被缓冲溶液，检测样品的包被体积为100μl，包被浓度为2μg/ml。
本发明试剂盒FGF23抗体与生物素标记试剂Sulfo-NHS-LC-Biontin标记条件为重量比为1∶5～1∶12，混合后的浓度为2μg/ml。
本发明试剂盒所述的casein盐溶液为FGF23、检测抗体和链霉素亲和素的稀释液，链霉素亲和素的稀释倍数是1∶3000。
本发明试剂盒所述的链霉素亲和素与底物的作用时间为10分钟。
本发明试剂盒FGF23的最低检测限值为1opg/ml，检测线性范围是2400pg/ml～1opg/ml。
本发明的有益效果为为慢性肾病的早期检测提供了一种体外检测试剂盒，该试剂盒对FGF23临床检测的灵敏度达到1opg/ml，比常规单抗提高了10倍的灵敏度，对于慢性肾病的早期诊断具有重要意义。
附图说明
图1是本发明实施例吸光度值与样品浓度间的线性关系。
具体实施方式
下面结合实施例对本发明做进一步的说明，以下所述，仅是对本发明的较佳实施例而已，并非对本发明做其他形式的限制，任何熟悉本专业的技术人员可能利用上述揭示的技术内容加以变更为同等变 化的等效实施例。凡是未脱离本发明方案内容，依据本发明的技术实质对以上实施例所做的任何简单修改或等同变化，均落在本发明的保护范围内。
实施例1FGF23检测试剂盒的制备
利用FGF23-Fc抗原免疫小鼠，获得杂交瘤细胞阳性FGF23抗体克隆，针对其中的1276株克隆进行了筛选，检测了他们对FGF23蛋白的结合特异性，筛选得到编号278、6B12和6H1的三种抗体。
利用前期筛选的三株FGF23单克隆抗体278，6B12和6H1进行FGF23ELISA检测试剂盒检测条件的优化。
1)2mg/ml 6B12和6H1各100μl按1∶20、1∶10、1∶5室温进行30分钟生物素化反应，反应结束用等体积1％BSA结束反应。用包被抗体278，进行包被，3ng/ml FGF23三倍梯度稀释，生物素化抗体进行检测，考察生物素化效率，生物素试剂选择NHS-PEG4-Biontin(Lot#ND 172078)，具体情况见表1：
表1 生物素标记比率优化
Figure PCTCN2015097926-appb-000001
由表1可知抗体6B12进行生物素标记的最佳比例是1∶5；6H1进行生物素化标记的最佳浓度是1∶10。
根据以上实验结果，围绕最佳生物素化条件，进行生物标记条件优化，6B12以1∶8、1∶6、1∶5、1∶4进行生物素化反应；6H1以1∶12、1∶10、1∶8进行标记。
表2 生物素标记比率优化
Figure PCTCN2015097926-appb-000002
表2研究数据表明：抗体6B12进行生物素标记的最佳比例是1∶8；6H1进行生物素化标记的最佳浓度是1∶10。
根据以上实验结果，围绕最佳生物素化条件，进行生物标记条件优化，6B12以1∶7、1∶8、1∶9进行生物素化反应；6H1以1∶9、1∶10、1∶11进行标记。表3研究数据表明：抗体6B12进行生物素标记的最佳比例是1∶8；6H1进行生物素化标记的最佳浓度是1∶10和1∶9。
表3 生物素标记比率优化
Figure PCTCN2015097926-appb-000003
Figure PCTCN2015097926-appb-000004
2)选择Thermo scientific公司生产的两种生物素试剂：Sulfo-NHS-LC-Biontin(Lot#21327)和NHS-PEG4-Biontin(Lot#ND172078)研究标记灵敏的差异，实验结果见表4和表5。上述试验数据，综合考察灵敏度和本底背景，选择Sulfo-NHS-LC-Biontin(Lot#21327)为最终生物素标记试剂。
表4 Sulfo-NHS-LC-Biontin标记数据
Figure PCTCN2015097926-appb-000005
表5 NHS-PEG4-Biontin标记数据
Figure PCTCN2015097926-appb-000006
3)Sulfo-NHS-LC-Biontin(Lot#21327)生物素试剂最佳标记条件优化
根据表4试验数据，围绕6B12和6H1最佳灵敏度标记数据，进行标记比例细化，6B12以1∶8，1∶9，1∶10，1∶11，1∶12，1∶20进行生物素化反应；6H1以1∶3，1∶4，1∶5，1∶6，1∶7，1∶10进行标记，实验数据见表6。根据表6试验数据，围绕6B12和6H1最佳灵敏度标记数据，进行标记比例细化，6B12和6H1分别以1∶8，1∶9，1∶10，1∶11，1∶12进行生物素化反应，实验数据见表7。表7实验数据说明，6B12选择1∶8进行标记，6H1选择1∶10进行标记时，检测灵敏度最高。
表6 6B12和6H1最佳生物素标记比率优化
Figure PCTCN2015097926-appb-000007
表7 6B12和6H1生物素标记比率优化
Figure PCTCN2015097926-appb-000008
Figure PCTCN2015097926-appb-000009
4)检测抗体6B12，6H1以及两种抗体混合后，分别以1μg/ml和2μg/ml进行测试，实验数据见表8。
表8 检测抗体最佳检测浓度筛选
Figure PCTCN2015097926-appb-000010
5)包被缓冲溶液的选择
分别用0.1M，pH 7.0磷酸盐缓冲溶液，以及pH 9.0，0.1M硼氢化钠溶液作为包被溶液考察包被后检测灵敏度以及本底水平，实验数据见表9。表9实验数据说明，综合考虑检测灵敏度和本底水平，选择pH 7.0磷酸盐缓冲溶液作为FGF23试剂盒的包被溶液。
表9 磷酸盐缓冲溶液和硼氢化钠溶液包被效果比较
Figure PCTCN2015097926-appb-000011
6)包被体积对检测灵敏度及本底水平的影响
包被100μl和200μl anti-FGF23，使用不同标记比率的检测 抗体，考察FGF23试剂盒检测灵敏度和本底水平，筛选最佳包被体积，实验数据见表10。表10数据说明，增加包被体积，可提高检测灵敏度，但亦相应增加了检测中的本底水平，相对于灵敏度而言，本底水平对临床样品的检出影响更大，因此综合考虑选择100μl的包被体积用于anti-FGF23的包被体积。
表10 不同包被体积对FGF23试剂盒检测灵敏度和本底水平影响
Figure PCTCN2015097926-appb-000012
7)Anti-FGF23包被浓度对检测灵敏度和本底水平的影响
选择2μg/ml，4μg/ml，8μg/ml，16μg/ml，20μg/ml作为anti-FGF23的包被浓度，考察各不同包被浓度对FGF23检测试剂盒检测灵敏度和本底水平的影响，实验数据见表11。
表11 不同anti-FGF23浓度对FGF23检测试剂盒灵敏度和本底水平影响
Figure PCTCN2015097926-appb-000013
表11数据说明，增加anti-FGF23的包被浓度，检测灵敏度增加不显著，但本底水平有相应提高，因此选择2μg/ml作为anti-FGF23 的包被浓度。
8)酶浓度及酶与底物作用时间对检测灵敏度的影响
选择链霉素亲和素稀释度1∶2000和1∶3000，链霉素亲和素与TMB作用时间为5分钟和10分钟，考核酶浓度和酶与底物作用时间对检测灵敏度和本底水平的影响，实验数据见表12。表12数据说明链霉素亲和素稀释浓度1∶2000和1∶3000对FGF23试剂盒检测灵敏度影响不显著，酶促反应10分钟可增加FGF23检测灵敏度，且本底水平增加不显著，因此选择1∶3000作为酶反应浓度，酶与底物作用10分钟作为酶反应时间。
表12 链霉素亲和素稀释度及酶与底物作用时间对FGF23试剂盒灵敏度影响
Figure PCTCN2015097926-appb-000014
9)样品稀释液对本底水平的影响
选择casein，0.1M PBS与0.05％tween20混合物，1％BSA与0.05％tween混合物作为FGF23、检测抗体和酶的稀释液，研究不同样品稀释液对本底水平的影响，实验数据见表13。
表13 不同样品稀释液对检测本底水平的影响
Figure PCTCN2015097926-appb-000015
Figure PCTCN2015097926-appb-000016
表13数据说明，以casein盐做样品稀释液本底水平最低，后续实验还将针对这部分工作进行深入研究。
综上所述，该试剂盒生物素标记试剂选择为Sulfo-NHS-LC-Biontin(Lot#21327)，标记条件为6B12选择1∶8进行标记，6H1选择1∶10进行标记，生物素标记抗体选择6B12和6H1两种抗体混合，检测浓度为2μg/ml；使用pH 7.0，0.1M PBS作为包被缓冲溶液，anti-FGF23的包被体积是100μl，anti-FGF23的包被浓度是2μg/ml，样品稀释液选择casein盐溶液，链霉素亲和素的稀释倍数是1∶3000，酶与底物的作用时间是10分钟；
实施例2最低检测限值的研究
针对实施例1试剂盒，FGF23以10000pg/ml和2400g/ml开始进行3倍梯度稀释，考核最低检测浓度与空白对照孔间p值是否≤0.05，作为FGF23的最低检测限值。表14和表15。
表14 FGF23试剂盒最低检测限制研究
Figure PCTCN2015097926-appb-000017
Figure PCTCN2015097926-appb-000018
表15 FGF23试剂盒最低检测限制研究
Figure PCTCN2015097926-appb-000019
表14和15数据说明，以casein盐做样品稀释液本底水平最低实验数据表明，FGF23试剂盒最低检测限值为10pg/ml。
试剂盒FGF23标准品浓度以2400pg/ml起始，进行三倍梯度稀释，按上述优化条件进行包被，TMB进行显色，研究吸光度值与样品浓度间的线性关系。实验结果见图1实验数据表明，FGF23检测试剂盒的线性范围2400pg/ml～10pg/ml。

Claims (6)

  1. 一种慢性肾病诊断试剂盒，其特征在于：FGF23单抗隆抗体、生物素标记试剂Sulfo-NHS-LC-Biontin、缓冲溶液、casein盐溶液、链霉素亲和素和底物TMB。
  2. 根据权利要求1所述的诊断试剂盒，其特征在于：所述的缓冲溶液为pH 7.0、0.1M的PBS。
  3. 根据权利要求1所述的诊断试剂盒，其特征在于：所述的FGF23单抗隆抗体与生物素标记试剂Sulfo-NHS-LC-Biontin标记条件为重量比为1∶5～1∶12，混合后的浓度为2μg/ml。
  4. 根据权利要求1所述的诊断试剂盒，其特征在于：所述的链霉素亲和素的稀释倍数为1∶3000，所使用的稀释液为casein盐溶液。
  5. 根据权利要求1所述的诊断试剂盒，其特征在于：所述的链霉素亲和素与底物的作用时间为10分钟。
  6. 根据权利要求1所述的诊断试剂盒，其特征在于：所述的试剂盒最低检测限值为10pg/ml，检测线性范围是2400pg/ml～10pg/ml。
PCT/CN2015/097926 2014-12-23 2015-12-18 一种慢性肾病诊断试剂盒 WO2016101847A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US15/300,236 US20170184593A1 (en) 2014-12-23 2015-12-18 A chronic renal disease diagnostic kit

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201410811167.6A CN104502596A (zh) 2014-12-23 2014-12-23 一种慢性肾病诊断试剂盒
CN201410811167.6 2014-12-23

Publications (1)

Publication Number Publication Date
WO2016101847A1 true WO2016101847A1 (zh) 2016-06-30

Family

ID=52944009

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2015/097926 WO2016101847A1 (zh) 2014-12-23 2015-12-18 一种慢性肾病诊断试剂盒

Country Status (3)

Country Link
US (1) US20170184593A1 (zh)
CN (1) CN104502596A (zh)
WO (1) WO2016101847A1 (zh)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018145119A1 (en) * 2017-02-06 2018-08-09 Astute Medical, Inc. Methods and compositions for diagnosis and prognosis of renal injury and renal failure
CN110568181A (zh) * 2019-09-12 2019-12-13 苏州普瑞斯生物科技有限公司 一种微量白蛋白免疫比浊法检测试剂盒及其制备方法
CN110763847A (zh) * 2019-11-07 2020-02-07 苏州普瑞斯生物科技有限公司 一种补体c3的检测试剂盒及制备方法

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104502596A (zh) * 2014-12-23 2015-04-08 温州医科大学 一种慢性肾病诊断试剂盒
CN109633176A (zh) * 2019-01-11 2019-04-16 广东医科大学附属医院 一种肾病基因治疗诊断试剂盒
CN112285356A (zh) * 2019-07-25 2021-01-29 苏州普瑞斯生物科技有限公司 α1-抗胰蛋白酶免疫比浊法检测试剂盒的制备方法

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1993618A (zh) * 2004-07-30 2007-07-04 株式会社先端生命科学研究所 探针复合物
WO2008089936A1 (en) * 2007-01-22 2008-07-31 Medizinische Universität Innsbruck Novel markers for chronic kidney disease
WO2009091556A2 (en) * 2008-01-17 2009-07-23 The General Hospital Corporation Diagnostic methods and kits using fibroblast growth factor-23
WO2012055096A1 (zh) * 2010-10-27 2012-05-03 温州医学院生物与天然药物开发中心有限公司 抗fgf-23抗体的制备方法及用途
CN104502596A (zh) * 2014-12-23 2015-04-08 温州医科大学 一种慢性肾病诊断试剂盒

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US16133A (en) * 1856-11-25 Apparatus for hoisting- coal
EP1466925B1 (en) * 2001-12-28 2009-09-02 Kyowa Hakko Kirin Co., Ltd. Antibodies against fibroblast growth factor 23
US20030198959A1 (en) * 2002-03-28 2003-10-23 Kurnit David M. Methods and compositions for analysis of urine samples in the diagnosis and treatment of kidney diseases
DE602005013288D1 (de) * 2004-12-20 2010-05-06 Antibodyshop As M lipocalin (ngal) als diagnostischer marker für nierenerkrankungen
CN101105498B (zh) * 2007-08-08 2011-06-01 中国人民解放军军事医学科学院卫生学环境医学研究所 人、大鼠、小鼠hsp70双抗夹心法检测试剂盒
CN101226196B (zh) * 2008-02-02 2012-05-09 南方医科大学 一种检测ⅱ型登革病毒ns1抗原的免疫诊断试剂盒
JP5981144B2 (ja) * 2009-02-06 2016-08-31 アスチュート メディカル，インコーポレイテッド 腎損傷および腎不全の診断および予後診断
EP3578979B1 (en) * 2010-08-31 2022-02-23 Minaris Medical Co., Ltd. Method for measuring fibroblast growth factor-23 and reagent therefor
TW201250248A (en) * 2011-04-25 2012-12-16 Kyowa Medex Co Ltd Prognostication method of renal failure

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1993618A (zh) * 2004-07-30 2007-07-04 株式会社先端生命科学研究所 探针复合物
WO2008089936A1 (en) * 2007-01-22 2008-07-31 Medizinische Universität Innsbruck Novel markers for chronic kidney disease
WO2009091556A2 (en) * 2008-01-17 2009-07-23 The General Hospital Corporation Diagnostic methods and kits using fibroblast growth factor-23
WO2012055096A1 (zh) * 2010-10-27 2012-05-03 温州医学院生物与天然药物开发中心有限公司 抗fgf-23抗体的制备方法及用途
CN104502596A (zh) * 2014-12-23 2015-04-08 温州医科大学 一种慢性肾病诊断试剂盒

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018145119A1 (en) * 2017-02-06 2018-08-09 Astute Medical, Inc. Methods and compositions for diagnosis and prognosis of renal injury and renal failure
CN110568181A (zh) * 2019-09-12 2019-12-13 苏州普瑞斯生物科技有限公司 一种微量白蛋白免疫比浊法检测试剂盒及其制备方法
CN110763847A (zh) * 2019-11-07 2020-02-07 苏州普瑞斯生物科技有限公司 一种补体c3的检测试剂盒及制备方法

Also Published As

Publication number Publication date
CN104502596A (zh) 2015-04-08
US20170184593A1 (en) 2017-06-29

Similar Documents

Publication Publication Date Title
WO2016101847A1 (zh) 一种慢性肾病诊断试剂盒
CN106568978A (zh) 血清淀粉样蛋白a检测方法及试剂
KR101032607B1 (ko) 간암 진단용 단백질성 마커
CA2882151C (en) Chemiluminescent protein chip, method and kit for detecting seroglycoid fucosylation index
JP5424702B2 (ja) 尿中ヒトメガリンを測定することを含む腎疾患検出方法
JP7166364B2 (ja) Ｃ反応性タンパク質を全域検出する方法及び対応するキット
US10429390B2 (en) Antibody cocktail systems and methods for classification of histologic subtypes in lung cancer
JP2011530514A5 (zh)
WO2016011852A1 (zh) 膀胱肿瘤相关抗原检测试剂盒
CN104949971B (zh) 肝癌早期诊断试剂盒及其应用
CN105492907B (zh) 新检测方法
JP2024056971A (ja) ＡＰＰ６６９－ｘのＮ末端を特異的に認識する抗体、及び免疫測定法
EP2815238B1 (en) A process for detection and optional quantification of an analyte
WO2016082445A1 (zh) 肽基精氨酸脱亚胺酶2在制备肿瘤临床诊断试剂中的应用
Jiang et al. The development of an indirect competitive immunomagnetic-proximity ligation assay for small-molecule detection
WO2016082444A1 (zh) 肽基精氨酸脱亚胺酶1在制备肿瘤临床诊断试剂中的应用
WO2018066409A1 (ja) 糖尿病性腎症の早期病態の特異的な診断を可能とする検査方法
WO2015116961A1 (en) Methods and compositions for assaying vitamin d
CN111198271B (zh) 一种用于甘油醛-3-磷酸脱氢酶检测的化学发光酶联免疫试剂盒
CN108802400B (zh) 一种检测复合物IgA1-α1MG的方法及无创检测肾组织损伤的试剂盒
US9523690B2 (en) Biomarkers for the diagnosis and/or prognosis of clear cell renal cell carcinoma
WO2016027697A1 (ja) ＴＲＡＣＰ－５ｂ（酒石酸抵抗性酸性フォスファターゼ５ｂ）に特異的なタンパク定量法
KR20100018804A (ko) 간암 진행 진단용 단백질성 마커
Trouvé et al. Surface plasmon resonance shows a gender difference in circulating annexin A5 in human
Maeda et al. Urinary carboxylesterase 5A fragment as an early diagnostic marker of cat chronic kidney disease

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15871915

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 15300236

Country of ref document: US

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 15871915

Country of ref document: EP

Kind code of ref document: A1