Sealing reagent for improving suspension stability of latex microspheres, method and kit
A latex and microsphere technology, applied in the field of medical devices, can solve the problems of reagent failure, insufficient to ensure stable storage of microsphere components, microsphere aggregation, etc.
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Embodiment 1
[0156] Synthesis and characterization of embodiment 1 PS-AA oligomer
[0157] Add 30mL of pure water into a four-neck flask equipped with a stirrer, a thermometer and a dropping funnel, heat it to 65°C, and slowly add a mixture of 5g of styrene (Styrene, referred to as S) and 0.55g of isobutane and 10mL contains 0.5gNaHSO 3 , 1g (NH 4 ) 2 S 2 o 4 , 1.5g aqueous solution of isopropanol; control the rate of addition, so that the monomer and initiator solution are dripped within 30 minutes; after 1 hour of reaction, the temperature is raised to 80°C, and 25g of acrylic acid (Acrylic acid, referred to as AA) is slowly added dropwise, and the reaction is continued After 5 hours, a solid foamy product was formed at this time; the solid was collected by filtration and vacuum-dried for 24 hours; the polymer was taken out and ground into a fine powder to obtain a PS-AA copolymer with a molecular weight of about 2500-15000.
[0158] Infrared spectrum such as figure 1 As shown, the...
Embodiment 2
[0161] Example 2 Anti-FK506 Monoclonal Antibody Coupling with Carboxylated PS Microspheres and Microsphere Blocking
[0162] Add 60mg surface carboxy-modified latex microspheres (325nm, JSR), 250.0mg NHS and 10.0mg EDCI to 6mL MES buffer successively, activate microsphere carboxyl groups for 25min (25°C); centrifuge at 12000rpm for 30min, discard the supernatant , the microspheres were dispersed with 6ml of 10mM PBS (pH7.5) to form a suspension containing latex microspheres.
[0163] In the suspension, slowly drop 4.75mg anti-FK506 monoclonal antibody (Shanghai Yunze Biotechnology Co., Ltd., Feng-Bo Wu, Yun-Yun Yang, Xue-Bin Wang, et al.A sample processing method forimmunoassay of whole blood tacrolimus. Analytical Biochemistry, 2019, 576, 13–19), placed on a rotator for 24 hours (25°C); centrifuged at 12,000 rpm for 30 minutes, and removed the supernatant. Add 6 mL of TSA-polysorbate-20 (50 mM Tris-HCl, pH 7.5, containing 0.05% polysorbate-20, 0.05% NaN 3 ) to wash the micr...
Embodiment 3
[0170] Example 3 Effects of using different blocking solutions on the storage stability of microspheres
[0171] The latex microspheres coupled with anti-FK506 monoclonal antibody blocked by the above six different blocking solutions (1#, 2#, 3#, 4#, 5#, 6#) were stored at 2-8°C for 1, At 3, 5, 7, 9, 12, and 15 months, the particle size and particle size distribution of the microspheres were detected with a nanoparticle size analyzer (Nanotrac Wave II).
[0172] The result is as Figure 4-5 And as shown in Table 2. Figure 4 is the particle size distribution diagram of the initial microspheres (latex microspheres coupled with anti-FK506 monoclonal antibody before blocking). Figure 5 It is the particle size distribution diagram of microspheres sealed by 1#-6# blocking solution and stored for 15 months.
[0173] Table 2 is the microsphere dispersion property after 15 months storage of the microsphere suspension sealed by 6 kinds of blocking solutions
[0174] Table 2
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