WO2019148837A1 - 一种用于化学发光免疫检测的磁颗粒试剂及试剂盒 - Google Patents

一种用于化学发光免疫检测的磁颗粒试剂及试剂盒 Download PDF

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WO2019148837A1
WO2019148837A1 PCT/CN2018/104109 CN2018104109W WO2019148837A1 WO 2019148837 A1 WO2019148837 A1 WO 2019148837A1 CN 2018104109 W CN2018104109 W CN 2018104109W WO 2019148837 A1 WO2019148837 A1 WO 2019148837A1
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magnetic particle
particle reagent
chemiluminescence immunoassay
magnetic
protein
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PCT/CN2018/104109
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French (fr)
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张娜娜
徐滕
王晓燕
周慧林
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苏州长光华医生物医学工程有限公司
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/76—Chemiluminescence; Bioluminescence
    • 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
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54326—Magnetic particles
    • G01N33/54333—Modification of conditions of immunological binding reaction, e.g. use of more than one type of particle, use of chemical agents to improve binding, choice of incubation time or application of magnetic field during binding reaction
    • 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
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54326—Magnetic particles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2446/00—Magnetic particle immunoreagent carriers
    • G01N2446/80—Magnetic particle immunoreagent carriers characterised by the agent used to coat the magnetic particles, e.g. lipids
    • G01N2446/84—Polymer coating, e.g. gelatin

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  • the present application relates to a magnetic particle reagent and a kit for chemiluminescence immunoassay, and belongs to the technical field of chemiluminescence immunoassay.
  • magnetic particle chemiluminescence immunoassay is a novel labeling immunoassay technology that combines a chemiluminescent system with an immune reaction for detecting trace antigens or antibodies. It has the advantages of good selectivity, high sensitivity, strong specificity, no radioactive hazard, fast analysis speed, simple equipment, etc. It has been widely used in environmental, clinical, food, drug testing and other fields, and has become a research hotspot of immunoassay methods. development trend.
  • the magnetic particle chemiluminescence immunoassay kit has been widely used in the market, but the magnetic particle reagent of the existing kit has poor stability under adverse conditions such as high temperature, and the magnetic particles are easy to aggregate and cannot be quickly mixed, resulting in chemiluminescence immunoassay.
  • the experimental results show that the luminescence value is small, the repeatability is poor, the background value is high, and the actual consumption of the magnetic particle reagent is large, and the use cost is relatively increased, which is inconvenient for the detection of clinical samples.
  • the technical problem to be solved by the present application is to provide a technical problem for solving the problem that the magnetic particle reagent used in the chemiluminescence immunoassay is poor in high temperature and the like, and the magnetic particles are easy to aggregate and cannot be quickly mixed. Magnetic particle reagents and kits for chemiluminescence immunoassay.
  • a magnetic particle reagent for chemiluminescence immunoassay comprising 0.5-1.5 mg/mL streptavidin or anti-isothiocyanate antibody-coated magnetic particles, 0.5-5 mg/mL protein, 0.5- 10 mg/mL chloroacetamide-like protein protectant, 1-10 mg/mL preservative, 5-10 mg/mL polyhydroxy compound, 1-5 mg/mL surfactant buffer, the polyhydroxy compound is a polyol or polysaccharide
  • the surfactant is dodecyl polyethylene glycol ether (Brij35), polyethylene glycol to isooctylphenyl ether (TX-100), lauryl polyoxyethylene ether (flat plus O-20) At least one of them.
  • the polyol is at least one of glycerol, sorbitol or polyethylene glycol
  • the polyethylene glycol is preferably one of PEG 8000, PEG 10000, and PEG 20000.
  • the polysaccharide is at least one of fructose, sucrose, and trehalose.
  • the chloroacetamide-based protein protective agent is at least one of chloroacetamide, dichloropropenylamine, and oxaloacetone.
  • the streptavidin-coated magnetic particles are Fe 3 O 4 nanoparticles coated with dextran, and have a particle size of 0.1-5 ⁇ m.
  • the preservative is 2-methyl-4-isothiazolin-3-one, NaN 3 or proclin300, and the preservative has low toxicity and can effectively control microorganisms such as bacteria, fungi and yeast in the reagent. Growing.
  • the buffer is 50-100 mM, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer or Tris, having a pH of 7.2-7.6. Buffer.
  • HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
  • Tris having a pH of 7.2-7.6. Buffer.
  • the protein is at least one of bovine serum albumin (BSA), casein (Casein), gelatin (Gelatin).
  • BSA bovine serum albumin
  • casein casein
  • Gelatin gelatin
  • the present application also provides a chemiluminescent immunoassay kit comprising the above magnetic particle reagent, the kit further comprising an anti-test antigen antibody solution labeled with biotin or fluorescein isothiocyanate, and a chemiluminescent label-labeled anti-antibody Antigen antibody solution to be tested, chemiluminescent substrate solution, cleaning solution.
  • the magnetic particle reagent of the present application can stably and uniformly disperse magnetic particles coated with streptavidin or anti-isothiocyanate antibody after long-term placement, and can be easily aggregated, and can be applied to chemiluminescence immunoassay.
  • the magnetic particles are rapidly adsorbed by magnetic force, and have good preservation effect on streptavidin or anti-isothiocyanate antibody-coated magnetic particles, and can maintain streptavidin or antibiotic under adverse conditions such as high temperature.
  • the activity and stability of fluorescein isothiocyanate antibody and protein make the chemiluminescence value of chemiluminescence immunoassay large, reproducible, and small background value, specifically:
  • the polyhydroxy compound can form a hydrogen bond with streptavidin or an anti-isothiocyanate antibody, and the magnetic particles coated with streptavidin or anti-isothiocyanate antibody are adsorbed on the wall.
  • the magnetic particles are aggregated on the surface, and the combination of the polyhydroxy compound and the surfactant can effectively encapsulate the magnetic particles coated with streptavidin or anti-isothiocyanate antibody, and stably and uniformly dispersed in In the reagent, thereby improving the storage stability and dispersibility of the magnetic particle reagent.
  • the polyhydroxy compound cooperates with the protein to have a good sealing effect on the surface active sites of the magnetic particles, and reduces the non-specific adsorption of the chemiluminescent label-labeled antibody, thereby reducing the background value.
  • the chloroacetamide protein protectant can effectively protect the active site of streptavidin or anti-isothiocyanate antibody, and further ensure the activity of the protein and avoid the chain caused by adverse environmental factors such as high temperature transportation.
  • Mycomycin or anti-isothiocyanate antibodies and denaturation of proteins, polyols can further avoid the degradation of streptavidin or anti-isothiocyanate antibodies and proteins caused by high temperature transport.
  • the present application also provides a chemiluminescent immunoassay kit containing the above magnetic particle reagent, which has the advantages of long effective time, low cost, high precision and sensitivity of the detection result, good specificity, and is suitable for clinical detection.
  • Example 11 is a diagram showing precipitation of magnetic particles in a magnetic particle reagent of Example 11 of the present application
  • Figure 2 is a graph showing the precipitation of magnetic particles in the magnetic particle reagent of Comparative Example 1 of the present application
  • Figure 3 is a graph showing the precipitation of magnetic particles in the magnetic particle reagent of Comparative Example 2 of the present application.
  • Fig. 4 is a graph showing the precipitation of magnetic particles in the magnetic particle reagent of Comparative Example 3 of the present application.
  • Examples 1-11 and Comparative Examples 1-4 provide streptavidin coated magnetic particle reagents for chemiluminescent immunoassays containing components in the following amounts:
  • the magnetic particle reagents of the above Examples 1-11 and Comparative Examples 1-4 were divided into two parts and stored in an environment of 4 ° C and 37 ° C for 72 hours, respectively, and the above-mentioned magnetic particle reagent was used to adopt the AE-180 fully automatic chemiluminescence.
  • the analyzer detects the luminescence of alpha-fetoprotein in the blood sample.
  • the experimental steps are as follows:
  • S2 adding 20 ⁇ L of a magnetic particle reagent having a concentration of 0.8 mg/L, and reacting with the antigen-antibody complex of S1 for 10 minutes to form a magnetic antigen-antibody complex suspension;
  • S3 placing a magnetic antigen-antibody complex suspension of S2 in a magnetic field, washing the magnetic antigen-antibody complex;
  • S4 The washed magnetic antigen-antibody complex of S3 is reacted with 150 ⁇ L of a 0.1 mg/L acridinium ester-labeled anti-thyroglobulin secondary antibody (secondary antibody) solution for 20 minutes to form an antigen-antibody. - a second antimagnetic composite suspension;
  • S6 Inject 150 ⁇ L of chemiluminescence excitation solution A (0.1 M nitric acid and 1 g/100 mL of hydrogen peroxide) into the magnetic composite after washing with S5, and after 150 seconds, inject 150 ⁇ L of chemiluminescence excitation solution B (0.25 M sodium hydroxide and 0.1). g/100tritonx-100), collect the chemiluminescence photon intensity within 5 seconds, and the luminescence is shown in Table 1.
  • chemiluminescence excitation solution A 0.1 M nitric acid and 1 g/100 mL of hydrogen peroxide
  • Table 1 uses different magnetic particle reagents to detect the luminescence of alpha-fetoprotein in blood samples.
  • Example 11 The comparison results of Example 11 and Comparative Examples 1-3 further confirm the relationship between the polyol and the surfactant.
  • the magnetic particle reagents of Examples 1-11 and Comparative Example 1 were stored at 37 ° C for 72 h, and the thyroglobulin-negative blood sample and thyroglobulin concentration were measured by the chemiluminescence immunoassay method of Example 1 to be 20 IU/mL, 60 IU.
  • the luminescence values of blood samples of /mL, 200 IU/mL, 600 IU/mL, and 2000 IU/mL are shown in Table 2.
  • the magnetic particle reagent (Examples 1-11) of the present application can achieve a small background value, a large luminescence value, and a good linear relationship even when applied to a chemiluminescence immunoassay even after high temperature transportation.
  • the magnetic particle reagents of the above Examples 1-11 and Comparative Example 4 were naturally mixed, and then allowed to stand for 120 minutes. The precipitation of magnetic particles in the magnetic particle reagent was observed under a microscope. As a result, the magnetic particle reagents of Examples 1-11 were not precipitated. Phenomenon, Comparative Example 1 has a significant precipitation phenomenon, and the precipitation phenomenon of Comparative Example 2 and Comparative Example 3 is not as obvious as Comparative Example 1, and magnetic particles in the magnetic particle reagent of Example 11, Comparative Example 1, Comparative Example 2, and Comparative Example 3 The precipitation conditions are shown in Figure 1, Figure 2, Figure 3 and Figure 4, respectively. It can be seen that only the combination of polyhydroxy compound and surfactant can make streptavidin or anti-isothiocyanate antibody coated. The magnetic particles are stably and uniformly dispersed in the reagent.

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Abstract

本申请涉及一种用于化学发光免疫检测的磁颗粒试剂及试剂盒，所述磁颗粒试剂包含有0.5-1.5mg/mL链霉亲和素或抗异硫氰酸荧光素抗体包被的磁颗粒、0.5-5mg/mL蛋白、0.5-10mg/mL氯乙酰胺类蛋白保护剂、1-10mg/mL防腐剂、5-10mg/mL多羟基化合物、1-5mg/mL表面活性剂的缓冲液，所述多羟基化合物为多元醇或多糖，所述表面活性剂为Brij35、TX-100、平平加O-20中的至少一种，本申请的磁颗粒试剂可使磁颗粒长期稳定、均匀地分散，且在高温等不利环境下能保存链霉亲和素或抗异硫氰酸荧光素抗体以及蛋白的活性和稳定性，使化学发光免疫分析的化学发光值大，重复性好，背景值小。

Description

一种用于化学发光免疫检测的磁颗粒试剂及试剂盒 技术领域
本申请涉及一种用于化学发光免疫检测的磁颗粒试剂及试剂盒，属于化学发光免疫分析技术领域。
背景技术
近年来，在免疫分析领域中免疫分析方法多种多样，其中磁微粒化学发光免疫分析法是将化学发光体系与免疫反应相结合，用于检测微量抗原或抗体的一种新型标记免疫测定技术，它具有选择性好、灵敏度高、特异性强、无放射性危害、分析速度快、设备简单等优点，在环境、临床、食品、药物检测等领域得到了广泛应用，成为免疫分析方法的研究热点和发展趋势。
目前磁微粒化学发光免疫检测试剂盒已广泛应用于市场，但现有试剂盒的磁颗粒试剂在高温等不利环境下的稳定性差，磁颗粒易聚集而又无法快速混匀，导致化学发光免疫检测实验的检测结果发光值小、重复性差、背景值高，且磁颗粒试剂的实际消耗多，使用成本相对增加，不便于临床样本的检测。
申请内容
本申请要解决的技术问题是：为解决现有用于化学发光免疫检测的磁颗粒试剂高温等不利环境下的稳定性差，磁颗粒易聚集而又无法快速混匀的技术问题，提供一种用于化学发光免疫检测的磁颗粒试剂及试剂盒。
本申请解决其技术问题所采用的技术方案是：
一种用于化学发光免疫检测的磁颗粒试剂，包含有0.5-1.5mg/mL链霉亲和素或抗异硫氰酸荧光素抗体包被的磁颗粒、0.5-5mg/mL蛋白、0.5-10mg/mL氯乙酰胺类蛋白保护剂、1-10mg/mL防腐剂、5-10mg/mL多羟基化合物、1-5mg/mL表面活性剂的缓冲液，所述多羟基化合物为多元醇或多糖，所述表面活性剂为十二烷基聚乙二醇醚(Brij35)、聚乙二醇对异辛基苯基醚(TX-100)、月桂醇聚氧乙烯醚(平平加O-20)中的至少一种。
优选地，所述多元醇为丙三醇、山梨醇或聚乙二醇中的至少一种，所述聚乙二醇优选为PEG8000、PEG10000、PEG20000中的一种。
优选地，所述多糖为果糖、蔗糖、海藻糖中的至少一种。
优选地，所述氯乙酰胺类蛋白保护剂为氯乙酰胺、二氯丙烯胺、解草酮中的至少一种。
优选地，所述链霉亲和素包被的磁颗粒为包裹有葡聚糖的Fe 3O 4纳米颗粒，粒径大小为0.1-5μm。
优选地，所述防腐剂为2-甲基-4-异噻唑啉-3-酮、NaN 3或proclin300，所述防腐剂的毒性低，能有效控制试剂中细菌、真菌和酵母菌等微生物的生长。
优选地，所述缓冲液为50-100mM、PH为7.2-7.6的4-(2-羟乙基)-1-哌嗪乙磺酸(HEPES)缓冲液或三羟甲基氨基甲烷(Tris)缓冲液。
优选地，所述蛋白为牛血清白蛋白(BSA)、酪蛋白(Casein)、 明胶(Gelatin)中的至少一种。
本申请还提供一种含有上述磁颗粒试剂的化学发光免疫检测试剂盒，所述试剂盒还包括标记生物素或异硫氰酸荧光素的抗待测抗原抗体溶液，化学发光标记物标记的抗待测抗原抗体溶液，化学发光底物液，清洗液。
本申请的有益效果是：
本申请的磁颗粒试剂可使链霉亲和素或抗异硫氰酸荧光素抗体包被的磁颗粒在长期放置后稳定、均匀地分散而不易聚集，在应用于化学发光免疫分析时可以将磁颗粒被磁力快速吸附，且对链霉亲和素或抗异硫氰酸荧光素抗体包被的磁颗粒具有良好的保存效果，能在高温等不利环境下能保持链霉亲和素或抗异硫氰酸荧光素抗体以及蛋白的活性和稳定性，使化学发光免疫分析的化学发光值大，重复性好，背景值小，具体是：
(1)多羟基化合物可与链霉亲和素或抗异硫氰酸荧光素抗体形成氢键，降低链霉亲和素或抗异硫氰酸荧光素抗体包被的磁颗粒吸附于器壁表面而造成的磁颗粒聚集，多羟基化合物与表面活性剂的配合使用可将链霉亲和素或抗异硫氰酸荧光素抗体包被的磁颗粒有效地包裹起来，稳定、均匀地分散于试剂中，从而提高磁颗粒试剂保存稳定性和分散性。
(2)多羟基化合物与蛋白配合，可以对磁颗粒的表面活性位点起到良好的封闭效果，降低化学发光标记物标记抗体的非特异性吸附，从而降低了背景值。
(3)氯乙酰胺类蛋白保护剂可以有效地保护链霉亲和素或抗异硫氰酸荧光素抗体的活性位点，还可进一步保证蛋白活性，避免高温运输等不利环境因素导致的链霉亲和素或抗异硫氰酸荧光素抗体以及蛋白的变性，多羟基化合物可进一步避免高温运输导致的链霉亲和素或抗异硫氰酸荧光素抗体以及蛋白的变性。
另外，本申请还提供一种含有上述磁颗粒试剂的化学发光免疫检测试剂盒，使用的有效时间长，成本低，检测结果的精密度和灵敏度高，特异性好，适于临床检测的应用。
附图说明
下面结合附图和实施例对本申请进一步说明。
图1是本申请实施例11的磁颗粒试剂中磁颗粒的沉淀情况图；
图2是本申请对比例1的磁颗粒试剂中磁颗粒的沉淀情况图；
图3是本申请对比例2的磁颗粒试剂中磁颗粒的沉淀情况图；
图4是本申请对比例3的磁颗粒试剂中磁颗粒的沉淀情况图。
具体实施方式
现对本申请作进一步详细的说明。
实施例1-11和对比例1-4
实施例1-11和对比例1-4提供了用于化学发光免疫检测的链霉亲和素包被的磁颗粒试剂，所述磁颗粒试剂含有如下含量的组分：
Figure PCTCN2018104109-appb-000001
Figure PCTCN2018104109-appb-000002
Figure PCTCN2018104109-appb-000003
效果例1
将上述实施例1-11和对比例1-4的磁颗粒试剂均分成两份，分别置于4℃和37℃环境下保存72h后，利用上述磁颗粒试剂，采用AE-180全自动化学发光分析仪检测血样中甲胎蛋白的发光情况，实验步骤如下：
S1：将50μL血样和100μL浓度为0.4mg/L的标记生物素的抗甲状腺球蛋白抗体溶液反应10分钟，形成抗原-抗体复合体；
S2：加入20μL浓度为0.8mg/L的磁颗粒试剂，与S1的抗原-抗体复合体反应10分钟后，形成磁性抗原-抗体复合体悬浮液；
S3：将S2的磁性抗原-抗体复合体悬浮液置于磁场内，洗涤所述磁性抗原-抗体复合体；
S4：将S3的洗涤后的磁性抗原-抗体复合体与150μL浓度为0.1mg/L的吖啶酯标记的抗甲状腺球蛋白的第二抗体(二抗)溶液反应20分钟后，形成抗原-抗体-二抗磁性复合物悬浮液；
S5：将S4的磁性复合物悬浮液置于磁场内，洗涤所述磁性复合物；
S6：向S5洗涤后的磁性复合物中注入150μL化学发光激发液A(0.1M硝酸和1g/100mL过氧化氢)，1.5秒后，注入150μL化学发 光激发液B(0.25M氢氧化钠和0.1g/100tritonx-100)，收集5秒内的化学发光光子强度，发光情况如表1。
表1利用不同磁颗粒试剂检测血样中甲胎蛋白的发光情况
Figure PCTCN2018104109-appb-000004
备注：上述每个试验结果为重复测定样本10次的结果平均值。
由上表结果可知：(1)多羟基化合物与表面活性剂的配合使用可使链霉亲和素或抗异硫氰酸荧光素抗体包被的磁颗粒稳定、均匀地分散于试剂中，使得本申请的磁颗粒试剂(实施例1-11)应用于化学发光免疫分析的发光值大，重复性好，对比例1与实施例11的区别在于无多羟基化合物与表面活性剂，对比例2与实施例11的区别在于无多羟基化合物，对比例3与实施例11的区别在于无表面活性剂，实施例11和对比例1-3的对比结果进一步印证了多羟基化合物与表面活性剂对磁颗粒分散的协同作用；(2)氯乙酰胺类蛋白稳定剂可以有效地保护链霉亲和素或抗异硫氰酸荧光素抗体的活性位点，避免高 温等不利环境因素导致的链霉亲和素或抗异硫氰酸荧光素抗体的变性，对比例4与实施例11的区别在于无氯乙酰胺类蛋白稳定剂，实施例11的磁颗粒试剂置于4℃和37℃环境下保存72h后的发光值差别不大，而对比例4的磁颗粒试剂置于37℃环境下保存72h后比置于4℃下保存72h后的发光值降低很多，对比结果进一步印证了高温下氯乙酰胺类蛋白稳定剂对链霉亲和素或抗异硫氰酸荧光素抗体的活性的保护作用。
效果例2
将实施例1-11以及对比例1的磁颗粒试剂于37℃环境下保存72h后，通过效果例1的化学发光免疫分析方法检测甲状腺球蛋白阴性血样及甲状腺球蛋白浓度为20IU/mL、60IU/mL、200IU/mL、600IU/mL、2000IU/mL血样的发光值，结果见表2。
表2不同磁颗粒试剂检测不同浓度甲状腺球蛋白血样的发光值
Figure PCTCN2018104109-appb-000005
Figure PCTCN2018104109-appb-000006
由上表结果可知：本申请的磁颗粒试剂(实施例1-11)即使在高温运输后，应用于化学发光免疫分析的也能达到背景值小，发光值大，线性关系好的效果，使得测定结果的准确度高；通过实施例与对比例1-4的对比可以看出：(1)多羟基化合物与表面活性剂配合使用才可使链霉亲和素或抗异硫氰酸荧光素抗体包被的磁颗粒稳定、均匀地分散于试剂中，使磁颗粒试剂应用于化学发光免疫分析时的线性关系好；(2)多羟基化合物与蛋白配合可以有效降低发光背景值，若无多羟基化合物则对磁颗粒的表面活性位点的封闭效果欠佳，背景值明显升高；(3)氯乙酰胺类蛋白保护剂可以有效地保护链霉亲和素或抗异硫氰酸荧光素抗体的活性位点，还可避免高温运输等不利环境因素导致的链霉亲和素或抗异硫氰酸荧光素抗体以及蛋白的变性，若无氯乙酰胺类蛋白保护剂则会使磁颗粒试剂在高温运输后应用于化学发光免疫分析时的线性关系差，背景值高。
效果例3
将上述实施例1-11和对比例4的磁颗粒试剂自然混匀后，静置120min，显微镜下观察磁颗粒试剂中磁颗粒的沉淀情况，结果实施例1-11的磁颗粒试剂均无沉淀现象，对比例1有明显的沉淀现象，对比例2和对比例3的沉淀现象不如对比例1明显，实施例11、对比例1、对比例2和对比例3的磁颗粒试剂中磁颗粒的沉淀情况依次见图1、图2、图3和图4，由此可见，只有多羟基化合物与表面活性剂的配合使用才可使链霉亲和素或抗异硫氰酸荧光素抗体包被的磁颗粒稳定、均匀地分散于试剂中。
以上述依据本申请的理想实施例为启示，通过上述的说明内容，相关工作人员完全可以在不偏离本项申请技术思想的范围内，进行多样的变更以及修改。本项申请的技术性范围并不局限于说明书上的内容，必须要根据权利要求范围来确定其技术性范围。

Claims (9)

  1. 一种用于化学发光免疫检测的磁颗粒试剂，其特征在于，包含有0.5-1.5mg/mL链霉亲和素或抗异硫氰酸荧光素抗体包被的磁颗粒、0.5-5mg/mL蛋白、0.5-10mg/mL氯乙酰胺类蛋白保护剂、1-10mg/mL防腐剂、5-10mg/mL多羟基化合物、1-5mg/mL表面活性剂的缓冲液，所述多羟基化合物为多元醇或多糖，所述表面活性剂为十二烷基聚乙二醇醚(Brij35)、聚乙二醇对异辛基苯基醚(TX-100)、月桂醇聚氧乙烯醚(平平加O-20)中的至少一种。
  2. 根据权利要求1所述的用于化学发光免疫检测的磁颗粒试剂，其特征在于，所述多元醇为丙三醇、山梨醇或聚乙二醇中的至少一种，所述聚乙二醇优选为PEG8000、PEG10000、PEG20000中的一种。
  3. 根据权利要求1或2所述的用于化学发光免疫检测的磁颗粒试剂，其特征在于，所述多糖为果糖、蔗糖、海藻糖中的至少一种。
  4. 根据权利要求1-3任一项所述的用于化学发光免疫检测的磁颗粒试剂，其特征在于，所述氯乙酰胺类蛋白保护剂为氯乙酰胺、二氯丙烯胺、解草酮中的至少一种。
  5. 根据权利要求1-4任一项所述的用于化学发光免疫检测的磁颗粒试剂，其特征在于，所述链霉亲和素包被的磁颗粒为包裹有葡聚糖的Fe 3O 4纳米颗粒，粒径大小为0.1-5μm。
  6. 根据权利要求1-5任一项所述的用于化学发光 免疫检测的磁颗粒试剂，其特征在于，所述防腐剂为2-甲基-4-异噻唑啉-3-酮、NaN 3或proclin300。
  7. 根据权利要求1-6任一项所述的用于化学发光免疫检测的磁颗粒试剂，其特征在于，所述缓冲液为50-100mM、PH为7.2-7.6的4-(2-羟乙基)-1-哌嗪乙磺酸(HEPES)缓冲液或三羟甲基氨基甲烷(Tris)缓冲液。
  8. 根据权利要求1-7任一项所述的用于化学发光免疫检测的磁颗粒试剂，其特征在于，所述蛋白为牛血清白蛋白(BSA)、酪蛋白(Casein)、明胶(Gelatin)中的至少一种。
  9. 一种含有权利要求1-8任一项所述的磁颗粒试剂的化学发光免疫检测试剂盒，所述试剂盒还包括标记生物素或异硫氰酸荧光素的抗待测抗原抗体溶液，化学发光标记物标记的抗待测抗原抗体溶液，化学发光底物液，清洗液。
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