磁微粒化学发光免疫分析是将磁性分离技术、化学发光技术、免疫分析技术三者结合的分析方法。磁微粒作为载体偶联抗体或抗原,可捕获目标分子,因其具有超顺磁性可实现多次磁分离,有利于检测仪器的全自动化并加快检测速度。目前,磁微粒化学免疫分析技术因其灵敏度高、检测范围宽、自动化程度高等优点已被广泛应用于生物医学检测领域。对甲苯磺酰基磁珠表面有对甲苯磺酰基修饰,可与蛋白共价结合,表面具有丰富的活性基团,无需进一 步活化,即可在温和碱性条件下直接用于与多种生物配基 (抗体、活性蛋白和多肽等)的氨基或巯基的共价偶联,形成的亚胺结构稳定,不易降解;同时,对甲苯磺酰基磁珠兼具偶联工艺易操作,易放大的特点,是生物大分子固定化的理想选择。参数T1506717-1.2μmT1506717-2.8μm外观红棕色悬浊液红棕色悬浊液固含量10 mg/mL10 mg/mL粒径及分布1.2± 0.1 μm, CV2.8± 0.1 μm, CV表面基团含量40-80 μmol/g40-90 μmol/g产品特点1、弥散式结构:磁核材料为超顺磁性γ-Fe₂O₃纳米颗粒2、氧化铁含量:12%-20%(不同磁珠有差异)3、粒径:常规1.2、2.8μm4、 粒径高度均一,批间差CV5、分散性优良,检测本底低:自主创新设计的独特包覆层结构,具有极低的非特异性吸附6、单批次2公斤级产能:满足化学发光客户日常需求7、高稳定性:长时间存储后不会出现氧化变色和核磁泄露现象使用说明1、偶联条件选择(1) 偶联缓冲液不应含有伯胺基团,例如 Tris 缓冲液、甘氨酸缓冲液等。(2) 推荐偶联缓冲液为硼酸盐缓冲液(pH 9.5),此外,建议可尝试 pH 7.0(PBS)~9.5(硼酸盐),以筛选最佳偶联条件。(3) 以IgG抗体为例,推荐偶联比例为 IgG:Beads=20~30μg:1mg;但由于偶联蛋白的多样性,建议筛选不同的偶联比例,以获得最优结果。(4) 推荐偶联在37℃条件下进行,时长建议12h~18h。(5) 偶联完成需要进行封闭,封闭蛋白类型、用量建议进行筛选(常用BSA、酪蛋白等),推荐在37℃条件下封闭6~16h。2、使用注意事项(1) 磁性微球在取用前,务必充分振荡使磁珠呈均匀的悬浮状态。(2) 干燥、冷冻操作极易引起磁珠团聚,且会影响磁珠表面功能基团的活性,请注意保存条件。3、偶联推荐方案名称详述配制方法简述偶联缓冲液A0.1M 硼酸盐缓冲液, pH 9.50.618g H₃BO₃ (硼酸, MW 61.83),使用80mL纯化水溶解,用5M NaOH调节pH到9.5,补加纯化水至100mL偶联缓冲液B含3.0M硫酸铵的缓冲液A, pH 9.5 39.64g (NH₄)₂SO₄ (硫酸铵,MW 132.14),使用80mL偶联缓冲液A溶解,用5M NaOH调节pH到9.5,补加偶联缓冲液A至100mL封闭液50mM Tris+1.0% BSA+0.1% Tween 20, pH 7.40.606g Tris (三羟甲基氨基甲烷, MW 121.14),1.0g BSA,0.1g Tween20使用80mL纯化水溶解,调节pH到7.4,补加纯化水至100mL保存液50mM Tris+1.0% BSA+0.1% Tween 20, pH 7.4同上4、偶联方法(1) 使用涡旋混匀器将对甲苯磺酰基磁珠充分混匀悬浮,取10mg到离心管中;(2) 将离心管放置在磁力架上1~2min,移除上清液;(3) 加入1.0 mL的偶联缓冲液A,涡旋分散磁珠后,按照步骤 (2) 移除上清;(4) 重复 (3) 步骤2次;(5) 加入200μg抗体,用偶联缓冲液A补充至总体积为0.6mL,涡旋分散磁珠;(6) 加入0.4mL偶联缓冲液B,涡旋混匀;(7) 保持磁珠旋转混匀,在 37℃条件下16h后按照步骤 (2) 移除上清;(8) 加入1.0mL封闭液,涡旋分散磁珠,保持磁珠旋转混匀,在37°C条件下6~16h,按照步骤 (2) 移除上清;(9) 加入1.0mL保存液,涡旋分散磁珠后,按照步骤 (2) 移除上清;(10) 重复步骤 (9) 2次;(11) 加入1.0mL磁珠保存液,涡旋混匀,得到浓度为10mg/mL的磁珠混悬液,2~8℃下保存。应用实例1、T1506717-1.2μm在抗穆勒氏管激素(AMH)检测中与进口品牌D发光值对比结论:相同的磁珠偶联、试剂检测条件下,T1506717-1.2μm磁珠在AMH检测中发光值高于进口品牌D同类型1.0μm磁性微球;T1506717-1.2μm三批发光值差异极小,生产工艺稳定批间差小。2、T1506717-2.8μm在癌胚抗原(CEA)检测中与进口品牌D发光值对比结论:相同的磁珠偶联、试剂检测条件下,T1506717-2.8μm磁珠在CEA检测中发光值高于进口品牌D同类型2.8μm磁性微球;T1506717-2.8μm三批发光值差异极小,生产工艺稳定批间差小。
Magnetic Particle Chemiluminescence Immunoassay is an analytical method that combines magnetic separation technology, chemiluminescence technology, and immunoassay technology. Magnetic particles, as carriers conjugated with antibodies or antigens, can capture target molecules. Due to their superparamagnetic properties, they enable multiple magnetic separations, facilitating full automation of detection instruments and accelerating testing speed. Currently, magnetic particle chemiluminescence immunoassay technology is widely used in biomedical detection due to its high sensitivity, broad detection range, and high automation.Tosyl Magnetic Beads are surface-modified with tosyl groups, enabling covalent binding to proteins. The surface possesses abundant active groups, allowing direct covalent conjugation with various biological ligands (such as antibodies, active proteins, and peptides) via their amino or thiol groups under mild alkaline conditions, without requiring further activation. The resulting imine structure is stable and resistant to degradation. Additionally, tosyl magnetic beads feature an easy-to-operate and easily scalable conjugation process, making them an ideal choice for the immobilization of biomacromolecules.ParameterT1506717-1.2μmT1506717-2.8μmAppearanceRed-brown suspensionRed-brown suspensionSolid Content10 mg/mL10 mg/mLParticle Size & Distribution1.2± 0.1 μm, CV2.8± 0.1 μm, CVSurface Group Content40-80 μmol/g40-90 μmol/gProduct FeaturesDiffuse Structure: The magnetic core consists of superparamagnetic γ-Fe₂O₃ nanoparticles.Iron Oxide Content: 12% - 20% (varies depending on the specific bead type).Particle Size: Standard sizes of 1.2 μm and 2.8 μm.Highly Uniform Particle Size with CV < 5% between Batches: Ensures stable and reliable performance.Excellent Dispersion and Low Detection Background: Features a uniquely designed, proprietary coating layer that exhibits extremely low non-specific adsorption.Production Capacity of 2 kg per Batch: Meets the routine demands of chemiluminescence customers.High Stability: No oxidation/discoloration or magnetic core leakage occurs after long-term storage.Instructions for Use1. Conjugation Conditions(1) The conjugation buffer should not contain primary amine groups, such as Tris buffer or glycine buffer.(2) Borate buffer (pH 9.5) is recommended. Additionally, testing pH ranges from 7.0 (PBS) to 9.5 (borate) is suggested to determine optimal conditions.(3) For IgG antibodies, the recommended conjugation ratio is IgG:Beads = 20–30 μg:1 mg. However, due to the diversity of proteins, screening different ratios is advised for optimal results.(4) Conjugation is recommended at 37°C for 12–18 hours.(5) Blocking is required after conjugation. The type and amount of blocking protein should be optimized (commonly used agents include BSA, casein, etc.). Blocking at 37°C for 6–16 hours is recommended.2. Precautions(1) Before use, thoroughly vortex the magnetic beads to ensure a uniform suspension.(2) Drying or freezing can cause bead aggregation and may affect the activity of surface functional groups. Please adhere to the recommended storage conditions.3. Recommended Conjugation ProtocolItemDetailsPreparation MethodConjugation Buffer A0.1M Borate Buffer, pH 9.5Dissolve 0.618 g H₃BO₃ (Boric Acid, MW 61.83) in 80 mL purified water. Adjust pH to 9.5 with 5M NaOH, then add purified water to 100 mL.Conjugation Buffer BBuffer A containing 3.0M Ammonium Sulfate, pH 9.5Dissolve 39.64 g (NH₄)₂SO₄ (Ammonium Sulfate, MW 132.14) in 80 mL Conjugation Buffer A. Adjust pH to 9.5 with 5M NaOH, then add Conjugation Buffer A to 100 mL.Blocking Buffer50 mM Tris + 1.0% BSA + 0.1% Tween 20, pH 7.4Dissolve 0.606 g Tris (MW 121.14), 1.0 g BSA, and 0.1 g Tween 20 in 80 mL purified water. Adjust pH to 7.4, then add purified water to 100 mL.Storage Buffer50 mM Tris + 1.0% BSA + 0.1% Tween 20, pH 7.4Same as above.4. Conjugation Method(1) Thoroughly vortex the tosyl magnetic beads to suspend evenly, and transfer 10 mg to a centrifuge tube.(2) Place the tube on a magnetic rack for 1–2 minutes and remove the supernatant.(3) Add 1.0 mL of Conjugation Buffer A, vortex to disperse the beads, and remove the supernatant as in step (2).(4) Repeat step (3) twice.(5) Add 200 μg of antibody and supplement with Conjugation Buffer A to a total volume of 0.6 mL. Vortex to disperse the beads.(6) Add 0.4 mL of Conjugation Buffer B and vortex to mix.(7) Maintain the beads in suspension with mixing and react at 37°C for 16 hours. Remove the supernatant as in step (2).(8) Add 1.0 mL of Blocking Buffer, vortex to disperse the beads. Maintain suspension with mixing and block at 37°C for 6–16 hours. Remove the supernatant as in step (2).(9) Add 1.0 mL of Storage Buffer, vortex to disperse, and remove the supernatant as in step (2).(10) Repeat step (9) twice.(11) Add 1.0 mL of Storage Buffer, vortex to mix, resulting in a magnetic bead suspension at 10 mg/mL. Store at 2–8°C.Application Examples1. Comparison of Luminescence Values between T1506717-1.2μm and Imported Brand D in Anti-Müllerian Hormone (AMH) DetectionConclusion:Under identical bead conjugation and reagent detection conditions:The luminescence values of T1506717-1.2μm magnetic beads in AMH detection are higher than those of the similar 1.0μm magnetic microspheres from Imported Brand D.The luminescence values across three batches of T1506717-1.2μm show minimal variation, indicating a stable production process and low batch-to-batch variation.2. Comparison of Luminescence Values between T1506717-2.8μm and Imported Brand D in Carcinoembryonic Antigen (CEA) DetectionConclusion:Under identical bead conjugation and reagent detection conditions:The luminescence values of T1506717-2.8μm magnetic beads in CEA detection are higher than those of the similar 2.8μm magnetic microspheres from Imported Brand D.The luminescence values across three batches of T1506717-2.8μm show minimal variation, indicating a stable production process and low batch-to-batch variation.