技术领域technical field
本发明属于生物医用高分子材料领域，具体涉及一种基于蛋白二聚作用制备牛血清白蛋白纳米微球的方法。The invention belongs to the field of biomedical polymer materials, and in particular relates to a method for preparing bovine serum albumin nano-microspheres based on protein dimerization.
背景技术Background technique
基于天然高分子的纳米微球材料具有优异的生物相容性和可调的生物降解性，可用于药物、蛋白或基因的包埋和释放，目前已广泛用于药物控释、组织工程和再生医学等领域的研究。通常，药物在体内代谢较快，而频繁注射给药则会给病人带来极大的痛苦。选择合适的纳米载药体系，可实现药物在体内的缓慢释放，达到理想的治疗效果。在组织工程和再生医学领域，纳米微球常用于包埋细胞生长因子和基因，以提高细胞支架的生物活性，增加支架在体内的组织诱导能力。常用来制备纳米微球的天然高分子包括：牛血清白蛋白、明胶、壳聚糖、淀粉及纤维素等。Nano-microsphere materials based on natural polymers have excellent biocompatibility and adjustable biodegradability, and can be used for the embedding and release of drugs, proteins or genes, and have been widely used in drug controlled release, tissue engineering and regeneration Research in fields such as medicine. Usually, drugs are metabolized quickly in the body, and frequent injections will bring great pain to patients. Selecting an appropriate nano drug-carrying system can realize the slow release of drugs in the body and achieve the desired therapeutic effect. In the field of tissue engineering and regenerative medicine, nanospheres are often used to embed cell growth factors and genes to improve the biological activity of cell scaffolds and increase the tissue induction ability of scaffolds in vivo. Natural polymers commonly used to prepare nanospheres include: bovine serum albumin, gelatin, chitosan, starch, and cellulose.
基于天然高分子材料的纳米微球在制备工程中必须添加化学交联剂处理，其结构才能稳定。这些化学交联剂主要有多聚甲醛、戊二醛、水溶性碳化二亚胺和京尼平等，其作用是将天然高分子中的活性基团（如氨基、羧基和羟基等）经过缩合而达到交联的目的。虽然化学交联剂能使纳米微球在物理结构上比较稳定，但会导致包埋的活性蛋白类药物（如细胞生长因子等）变性而失活，不适合用于活性蛋白的传递；同时，化学交联剂具有细胞毒性，容易残留在材料中。如能避免使用有毒的化学交联剂，则有望得到高生物活性的纳米微球材料，这是降低材料毒性及提高药物输送效率的有效途径。 Nano-microspheres based on natural polymer materials must be treated with chemical cross-linking agents in the preparation process, so that their structures can be stabilized. These chemical cross-linking agents are mainly paraformaldehyde, glutaraldehyde, water-soluble carbodiimide and genipal, and their function is to condense the active groups in natural polymers (such as amino, carboxyl and hydroxyl, etc.) To achieve the purpose of cross-linking. Although the chemical cross-linking agent can make the nano-microspheres more stable in physical structure, it will cause the denaturation and inactivation of the embedded active protein drugs (such as cell growth factors, etc.), and is not suitable for the delivery of active proteins; at the same time, Chemical crosslinkers are cytotoxic and tend to remain in the material. If the use of toxic chemical cross-linking agents can be avoided, nano-microsphere materials with high biological activity are expected to be obtained, which is an effective way to reduce material toxicity and improve drug delivery efficiency. the
发明内容Contents of the invention
本发明的目的是提供一种制备牛血清白蛋白纳米微球的方法，避免使用有毒的化学交联剂，能保证材料的使用安全性和被包埋药物的生物活性。The purpose of the present invention is to provide a method for preparing bovine serum albumin nano-microspheres, which avoids the use of toxic chemical cross-linking agents and can ensure the safety of materials and the biological activity of embedded drugs.
实现本发明目的的技术解决方案为：The technical solution that realizes the object of the present invention is:
一种牛血清白蛋白纳米微球的制备方法，采用双端马来酰亚胺化聚乙二醇为交联分子，通过蛋白二聚作用及反相溶液法交联制备牛血清白蛋白，包括以下步骤：A method for preparing bovine serum albumin nano-microspheres, using double-ended maleimidated polyethylene glycol as a cross-linking molecule, and preparing bovine serum albumin through protein dimerization and reverse-phase solution method cross-linking, including The following steps:
步骤1、室温下将牛血清白蛋白溶于氯化钠溶液中；Step 1, dissolving bovine serum albumin in sodium chloride solution at room temperature;
步骤2、氩气保护下加入双端马来酰亚胺化聚乙二醇溶液；Step 2, adding double-ended maleimidated polyethylene glycol solution under the protection of argon;
步骤3、室温搅拌下滴加氢氧化钠溶液，调节pH；Step 3, adding sodium hydroxide solution dropwise under stirring at room temperature to adjust the pH;
步骤4、室温搅拌下滴加反相溶剂，至溶液混浊后停止搅拌；Step 4. Add the reverse phase solvent dropwise under stirring at room temperature, and stop stirring until the solution is cloudy;
步骤5、高速离心得到纳米微球，洗涤、冷冻、干燥后得到牛血清白蛋白纳米微球。Step 5, high-speed centrifugation to obtain nano-microspheres, washing, freezing, and drying to obtain bovine serum albumin nano-microspheres.
其中，步骤1中所述牛血清白蛋白在氯化钠溶液中的质量浓度为2~5%。Wherein, the mass concentration of bovine serum albumin in the sodium chloride solution described in step 1 is 2-5%.
步骤2中所述双端马来酰亚胺化聚乙二醇溶液的质量浓度为0.4~1%。The mass concentration of the double-end maleimidated polyethylene glycol solution described in step 2 is 0.4-1%.
步骤3中加入双端马来酰亚胺化聚乙二醇后，混合溶液pH值调节至7~9。After adding double-ended maleimidated polyethylene glycol in step 3, the pH value of the mixed solution is adjusted to 7-9.
步骤4中加入滴加的反相溶剂为丙酮或无水乙醇，滴加量为混合溶液体积的6~8倍。The reverse-phase solvent added dropwise in step 4 is acetone or absolute ethanol, and the dropwise amount is 6 to 8 times the volume of the mixed solution.
步骤5中先后用蒸馏水、无水乙醇洗涤。In step 5, wash with distilled water and absolute ethanol successively.
the
本发明采用双端马来酰亚胺化聚乙二醇为交联分子，在一定条件下能使牛血清白蛋白发生二聚作用产生交联而得到稳定的纳米微球。双端马来酰亚胺化聚乙二醇分子结构中含有马来酰亚胺基，它能与牛血清白蛋白分子中的巯基发生迈克尔加成反应，同理含有马来酰亚胺基的其它水溶性聚合物材料亦可作为交联分子。The invention adopts double-terminal maleimidated polyethylene glycol as the cross-linking molecule, and under certain conditions, bovine serum albumin can be dimerized to generate cross-linking to obtain stable nanometer microspheres. The molecular structure of double-ended maleimidated polyethylene glycol contains maleimide groups, which can undergo Michael addition reaction with the sulfhydryl groups in bovine serum albumin molecules. Other water-soluble polymeric materials can also serve as crosslinking molecules.
本发明制备的牛血清白蛋白纳米微球分布均匀，平均直径可控制在27~88纳米。纳米微球的直径随着牛血清白蛋白溶液的浓度的增加而变大，而乙二醇浓度对纳米微球的尺寸影响不大。本发明所制备的多聚糖纳米微球结构稳定，在磷酸盐缓冲溶液中培养24小时后直径变化很小，适用于体内药物治疗。The bovine serum albumin nanometer microspheres prepared by the invention are evenly distributed, and the average diameter can be controlled at 27-88 nanometers. The diameter of nanospheres increases with the concentration of bovine serum albumin solution, while the concentration of ethylene glycol has little effect on the size of nanospheres. The polysaccharide nanometer microsphere prepared by the invention has a stable structure, little change in diameter after being cultured in the phosphate buffer solution for 24 hours, and is suitable for drug treatment in vivo.
原料和试剂：牛血清白蛋白，购于南京生兴生物技术有限公司；双端马来酰亚胺化聚乙二醇，购于上海西宝生物科技有限公司；丙酮，分析纯，上海凌峰化学试剂有限公司；磷酸氢二钠、无水乙醇，分析纯，成都科龙化工试剂厂。氯化钠、氯化钾、磷酸二氢钾、氢氧化钠，分析纯，南京化学试剂有限公司。磷酸盐缓冲溶液的配制：称取分析纯氯化钠8克、氯化钾0.2克、磷酸氢二钠2.9克、磷酸二氢钾0.2克，溶于1000毫升蒸馏水中。Raw materials and reagents: bovine serum albumin, purchased from Nanjing Shengxing Biotechnology Co., Ltd.; double-ended maleimidated polyethylene glycol, purchased from Shanghai Xibao Biotechnology Co., Ltd.; acetone, analytically pure, Shanghai Lingfeng Chemical Reagent Co., Ltd.; disodium hydrogen phosphate, absolute ethanol, analytical grade, Chengdu Kelong Chemical Reagent Factory. Sodium chloride, potassium chloride, potassium dihydrogen phosphate, sodium hydroxide, analytically pure, Nanjing Chemical Reagent Co., Ltd. Preparation of phosphate buffer solution: Weigh 8 grams of analytically pure sodium chloride, 0.2 grams of potassium chloride, 2.9 grams of disodium hydrogen phosphate, and 0.2 grams of potassium dihydrogen phosphate, and dissolve them in 1000 ml of distilled water.
纳米微球的粒径检测：纳米微球的直径通过纳米粒度分析仪（Malvern Zetasizer Nano ZS）测算。Particle size detection of nanospheres: the diameter of nanospheres is measured by a nanoparticle size analyzer (Malvern Zetasizer Nano ZS).
纳米微球的形貌检测：将干燥后的纳米微球喷金（Cressington 108 Auto），在扫描电子显微镜（JSM-6330F, JEOL）上观察微观形貌。Morphology detection of nano-microspheres: The dried nano-microspheres were sprayed with gold (Cressington 108 Auto), and the microscopic morphology was observed on a scanning electron microscope (JSM-6330F, JEOL).
本发明采用ANOVA方差分析法，显著差异值p设为≤0.05。The present invention adopts ANOVA variance analysis method, and the significant difference valuep is set to ≤0.05.
本发明与现有技术相比，其显著优点是：1）避免使用了有毒化学交联剂，所制备的纳米微球中不存在毒性残留，保证了纳米微球的安全性；2）所涉及的蛋白二聚作用基于牛血清白蛋白与聚乙二醇分子之间，不与被包埋活性药物发生化学反应，能有效保护药物的生物活性，特别适用于基因、活性蛋白类药物的包埋与传递；3）本发明工艺和设备简单、易行、操作安全，同时具有制备温度低、处理周期短等优点，适合商业化生产。Compared with the prior art, the present invention has the following remarkable advantages: 1) avoiding the use of toxic chemical cross-linking agents, and there is no toxic residue in the prepared nano-microspheres, which ensures the safety of the nano-microspheres; 2) the involved The protein dimerization is based on the interaction between bovine serum albumin and polyethylene glycol molecules. It does not chemically react with the embedded active drug and can effectively protect the biological activity of the drug. It is especially suitable for the embedding of genes and active protein drugs. and transfer; 3) The process and equipment of the present invention are simple, easy to operate, safe to operate, and have the advantages of low preparation temperature and short processing cycle, and are suitable for commercial production.
the
下面结合附图对本发明做进一步详细的描述。The present invention will be described in further detail below in conjunction with the accompanying drawings.
附图说明Description of drawings
图1本发明制备纳米微球的示意图。 Fig. 1 is a schematic diagram of preparing nano-microspheres in the present invention. ``
图2为本发明实施例1纳米微球的直径与牛血清白蛋白浓度的关系。Fig. 2 is the relationship between the diameter of the nano-microsphere and the concentration of bovine serum albumin in Example 1 of the present invention.
图3为本发明实施例1纳米微球的直径与聚乙二醇浓度的关系。Fig. 3 is the relationship between the diameter of nano-microspheres and the concentration of polyethylene glycol in Example 1 of the present invention.
图4为本发明实施例1纳米微球的直径与pH值的关系。Fig. 4 is the relationship between the diameter of the nano-microspheres and the pH value in Example 1 of the present invention.
图5为本发明实施例1纳米微球的直径分布在37oC磷酸盐缓冲溶液中孵育24小时后的变化（实线为孵育前，虚线为孵育后）。Figure 5 shows the changes in the diameter distribution of nanospheres in Example 1 of the present invention after incubation in phosphate buffered saline solution at 37o C for 24 hours (the solid line is before incubation, and the dotted line is after incubation).
具体实施方式Detailed ways
本发明提供的一种制备牛血清白蛋白纳米微球的方法，其步骤包括：A kind of method for preparing bovine serum albumin nano microsphere provided by the invention, its step comprises:
步骤1、配制浓度为10mM的氯化钠水溶液，将100~250mg牛血清白蛋白溶解于5ml氯化钠溶液；Step 1, preparation concentration is the sodium chloride aqueous solution of 10mM, and 100～250mg bovine serum albumin is dissolved in 5ml sodium chloride solution;
步骤2、将4~10mg双端马来酰亚胺化聚乙二醇溶解于1ml氯化钠溶液中，氩气保护下滴加到牛血清白蛋白溶液中，室温下搅拌2~4小时；Step 2. Dissolve 4-10 mg of double-ended maleimidated polyethylene glycol in 1 ml of sodium chloride solution, add it dropwise to the bovine serum albumin solution under argon protection, and stir at room temperature for 2-4 hours;
步骤3、室温搅拌下滴加浓度为1N的氢氧化钠溶液，调节混合溶液pH值为7~9；Step 3, add dropwise a sodium hydroxide solution with a concentration of 1N under stirring at room temperature, and adjust the pH value of the mixed solution to 7~9;
步骤4、室温搅拌下滴加6~8倍体积的丙酮或乙醇，至溶液混浊；Step 4. Add 6-8 volumes of acetone or ethanol dropwise with stirring at room temperature until the solution becomes turbid;
步骤5、高速离心得到纳米微球，随后分别用蒸馏水、无水乙醇清洗，最后冷冻干燥。Step 5, high-speed centrifugation to obtain nano-microspheres, followed by washing with distilled water and absolute ethanol respectively, and finally freeze-drying.
the
实施例1：Example 1:
（1）配制浓度为10mM的氯化钠水溶液，将100牛血清白蛋白溶解于5ml氯化钠溶液，制备示意图见图1，牛血清白蛋白浓度与纳米微球直径的关系见图2；(1) Prepare a sodium chloride aqueous solution with a concentration of 10mM, and dissolve 100% bovine serum albumin in 5ml sodium chloride solution. The schematic diagram of the preparation is shown in Figure 1, and the relationship between the concentration of bovine serum albumin and the diameter of the nanospheres is shown in Figure 2;
（2）将4mg双端马来酰亚胺化聚乙二醇溶解于1ml氯化钠溶液中，氩气保护下滴加到牛血清白蛋白溶液中，室温下搅拌2小时，制备示意图见图1，双端马来酰亚胺化聚乙二醇浓度与纳米微球直径的关系见图3；(2) Dissolve 4mg of double-ended maleimidated polyethylene glycol in 1ml of sodium chloride solution, add it dropwise into the bovine serum albumin solution under the protection of argon, and stir at room temperature for 2 hours. The schematic diagram of the preparation is shown in the figure 1. The relationship between the concentration of double-ended maleimidated polyethylene glycol and the diameter of nanospheres is shown in Figure 3;
（3）室温搅拌下滴加浓度为1N的氢氧化钠溶液，调节混合溶液pH值为7， pH值与纳米微球直径的关系见图4；(3) Add dropwise a sodium hydroxide solution with a concentration of 1N under stirring at room temperature, and adjust the pH value of the mixed solution to 7. The relationship between the pH value and the diameter of the nanospheres is shown in Figure 4;
（4）室温搅拌下滴加6倍体积的丙酮或乙醇，至溶液混浊；(4) Add 6 times the volume of acetone or ethanol dropwise with stirring at room temperature until the solution is cloudy;
（5）高速离心得到纳米微球，随后分别用蒸馏水、无水乙醇清洗，最后冷冻干燥得到牛血清白蛋白纳米微球。(5) High-speed centrifugation to obtain nanospheres, followed by washing with distilled water and absolute ethanol respectively, and finally freeze-drying to obtain bovine serum albumin nanospheres.
图5为所得纳米微球在37oC磷酸盐缓冲溶液中孵育24小时后的直径变化（实线为孵育前，虚线为孵育后），结果显示纳米微球在孵育前后，其直径变化很小，说明该纳米微球结构稳定，满足药物释放应用。Figure 5 shows the diameter change of the obtained nanospheres after incubation in 37o C phosphate buffer solution for 24 hours (the solid line is before incubation, and the dotted line is after incubation). The results show that the diameter of the nanospheres changes little before and after incubation , indicating that the structure of the nano-microsphere is stable and meets the application of drug release.
the
实施例2：Example 2:
（1）配制浓度为10mM的氯化钠水溶液，将150mg牛血清白蛋白溶解于5ml氯化钠溶液；(1) Prepare a sodium chloride aqueous solution with a concentration of 10mM, and dissolve 150mg bovine serum albumin in 5ml sodium chloride solution;
（2）将5mg双端马来酰亚胺化聚乙二醇溶解于1ml氯化钠溶液中，氩气保护下滴加到牛血清白蛋白溶液中，室温下搅拌2小时；(2) Dissolve 5 mg of double-ended maleimidated polyethylene glycol in 1 ml of sodium chloride solution, add dropwise to the bovine serum albumin solution under argon protection, and stir at room temperature for 2 hours;
（3）室温搅拌下滴加浓度为1N的氢氧化钠溶液，调节混合溶液pH值为7；(3) Add dropwise sodium hydroxide solution with a concentration of 1N under stirring at room temperature, and adjust the pH value of the mixed solution to 7;
（4）室温搅拌下滴加6倍体积的丙酮或乙醇，至溶液混浊；(4) Add 6 times the volume of acetone or ethanol dropwise with stirring at room temperature until the solution is cloudy;
（5）高速离心得到纳米微球，随后分别用蒸馏水、无水乙醇清洗，最后冷冻干燥得到牛血清白蛋白纳米微球。(5) High-speed centrifugation to obtain nanospheres, followed by washing with distilled water and absolute ethanol respectively, and finally freeze-drying to obtain bovine serum albumin nanospheres.
the
实施例3：Example 3:
（1）配制浓度为10mM的氯化钠水溶液，将150mg牛血清白蛋白溶解于5ml氯化钠溶液；(1) Prepare a sodium chloride aqueous solution with a concentration of 10mM, and dissolve 150mg bovine serum albumin in 5ml sodium chloride solution;
（2）将6mg双端马来酰亚胺化聚乙二醇溶解于1ml氯化钠溶液中，氩气保护下滴加到牛血清白蛋白溶液中，室温下搅拌3小时；(2) Dissolve 6 mg of double-ended maleimidated polyethylene glycol in 1 ml of sodium chloride solution, add dropwise to the bovine serum albumin solution under the protection of argon, and stir at room temperature for 3 hours;
（3）室温搅拌下滴加浓度为1N的氢氧化钠溶液，调节混合溶液pH值为8；(3) Add dropwise sodium hydroxide solution with a concentration of 1N under stirring at room temperature, and adjust the pH value of the mixed solution to 8;
（4）室温搅拌下滴加7倍体积的丙酮或乙醇，至溶液混浊；(4) Add 7 times the volume of acetone or ethanol dropwise with stirring at room temperature until the solution is cloudy;
（5）高速离心得到纳米微球，随后分别用蒸馏水、无水乙醇清洗，最后冷冻干燥得到牛血清白蛋白纳米微球。(5) High-speed centrifugation to obtain nanospheres, followed by washing with distilled water and absolute ethanol respectively, and finally freeze-drying to obtain bovine serum albumin nanospheres.
the
实施例4：Example 4:
（1）配制浓度为10mM的氯化钠水溶液，将150mg牛血清白蛋白溶解于5ml氯化钠溶液；(1) Prepare a sodium chloride aqueous solution with a concentration of 10mM, and dissolve 150mg bovine serum albumin in 5ml sodium chloride solution;
（2）将7mg双端马来酰亚胺化聚乙二醇溶解于1ml氯化钠溶液中，氩气保护下滴加到牛血清白蛋白溶液中，室温下搅拌3小时；(2) Dissolve 7 mg of double-ended maleimidated polyethylene glycol in 1 ml of sodium chloride solution, add dropwise to the bovine serum albumin solution under the protection of argon, and stir at room temperature for 3 hours;
（3）室温搅拌下滴加浓度为1N的氢氧化钠溶液，调节混合溶液pH值为8；(3) Add dropwise sodium hydroxide solution with a concentration of 1N under stirring at room temperature, and adjust the pH value of the mixed solution to 8;
（4）室温搅拌下滴加7倍体积的丙酮或乙醇，至溶液混浊；(4) Add 7 times the volume of acetone or ethanol dropwise with stirring at room temperature until the solution is cloudy;
（5）高速离心得到纳米微球，随后分别用蒸馏水、无水乙醇清洗，最后冷冻干燥得到牛血清白蛋白纳米微球。(5) High-speed centrifugation to obtain nanospheres, followed by washing with distilled water and absolute ethanol respectively, and finally freeze-drying to obtain bovine serum albumin nanospheres.
the
实施例5：Example 5:
（1）配制浓度为10mM的氯化钠水溶液，将200mg牛血清白蛋白溶解于5ml氯化钠溶液；(1) Prepare a sodium chloride aqueous solution with a concentration of 10mM, and dissolve 200mg bovine serum albumin in 5ml sodium chloride solution;
（2）将9mg双端马来酰亚胺化聚乙二醇溶解于1ml氯化钠溶液中，氩气保护下滴加到牛血清白蛋白溶液中，室温下搅拌4小时；(2) Dissolve 9 mg of double-ended maleimidated polyethylene glycol in 1 ml of sodium chloride solution, add dropwise to the bovine serum albumin solution under the protection of argon, and stir at room temperature for 4 hours;
（3）室温搅拌下滴加浓度为1N的氢氧化钠溶液，调节混合溶液pH值为9；(3) Add dropwise sodium hydroxide solution with a concentration of 1N under stirring at room temperature, and adjust the pH value of the mixed solution to 9;
（4）室温搅拌下滴加8倍体积的丙酮或乙醇，至溶液混浊；(4) Add 8 times the volume of acetone or ethanol dropwise with stirring at room temperature until the solution is cloudy;
（5）高速离心得到纳米微球，随后分别用蒸馏水、无水乙醇清洗，最后冷冻干燥得到牛血清白蛋白纳米微球。(5) High-speed centrifugation to obtain nanospheres, followed by washing with distilled water and absolute ethanol respectively, and finally freeze-drying to obtain bovine serum albumin nanospheres.
the
实施例6：Embodiment 6:
（1）配制浓度为10mM的氯化钠水溶液，将250mg牛血清白蛋白溶解于5ml氯化钠溶液；(1) Prepare a sodium chloride aqueous solution with a concentration of 10mM, and dissolve 250mg bovine serum albumin in 5ml sodium chloride solution;
（2）将10mg双端马来酰亚胺化聚乙二醇溶解于1ml氯化钠溶液中，氩气保护下滴加到牛血清白蛋白溶液中，室温下搅拌4小时；(2) Dissolve 10 mg of double-ended maleimidated polyethylene glycol in 1 ml of sodium chloride solution, add dropwise to the bovine serum albumin solution under the protection of argon, and stir at room temperature for 4 hours;
（3）室温搅拌下滴加浓度为1N的氢氧化钠溶液，调节混合溶液pH值为9；(3) Add dropwise sodium hydroxide solution with a concentration of 1N under stirring at room temperature, and adjust the pH value of the mixed solution to 9;
（4）室温搅拌下滴加8倍体积的丙酮或乙醇，至溶液混浊；(4) Add 8 times the volume of acetone or ethanol dropwise with stirring at room temperature until the solution is cloudy;
（5）高速离心得到纳米微球，随后分别用蒸馏水、无水乙醇清洗，最后冷冻干燥得到牛血清白蛋白纳米微球。(5) High-speed centrifugation to obtain nanospheres, followed by washing with distilled water and absolute ethanol respectively, and finally freeze-drying to obtain bovine serum albumin nanospheres.
the
以上实施例涵盖了最具代表性的实验数据。The above examples cover the most representative experimental data.
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310687518.2ACN104710630A (en) | 2013-12-17 | 2013-12-17 | Bovine serum albumin nano microsphere preparation method |
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310687518.2ACN104710630A (en) | 2013-12-17 | 2013-12-17 | Bovine serum albumin nano microsphere preparation method |
| Publication Number | Publication Date |
|---|---|
| CN104710630Atrue CN104710630A (en) | 2015-06-17 |
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310687518.2APendingCN104710630A (en) | 2013-12-17 | 2013-12-17 | Bovine serum albumin nano microsphere preparation method |
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| CN (1) | CN104710630A (en) |
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| CN108801736A (en)* | 2018-06-29 | 2018-11-13 | 闽南师范大学 | A kind of acetaldehyde crosslinking bovine serum albumin(BSA) and its preparation and application |
| CN109010804A (en)* | 2017-06-09 | 2018-12-18 | 理查德·C·K·延 | Albumin Nanosphere Formulations for Controlling Surgical Bleeding |
| CN111420067A (en)* | 2020-03-09 | 2020-07-17 | 西南交通大学 | A kind of composite microsphere nanocarrier and its preparation method and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1524880A (en)* | 2003-02-28 | 2004-09-01 | 中国科学院过程工程研究所 | A protein cross-linking agent and its cross-linking method |
| CN101732258A (en)* | 2008-11-19 | 2010-06-16 | 复旦大学附属华山医院 | Nano microsphere preparation used for chemotherapy of tumors and preparation method thereof |
| CN102688499A (en)* | 2012-06-05 | 2012-09-26 | 中国科学院过程工程研究所 | Albumin-polyethylene glycol-medicinal molecular conjugate |
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| CN109010804A (en)* | 2017-06-09 | 2018-12-18 | 理查德·C·K·延 | Albumin Nanosphere Formulations for Controlling Surgical Bleeding |
| CN108801736A (en)* | 2018-06-29 | 2018-11-13 | 闽南师范大学 | A kind of acetaldehyde crosslinking bovine serum albumin(BSA) and its preparation and application |
| CN108801736B (en)* | 2018-06-29 | 2021-07-30 | 闽南师范大学 | A kind of acetaldehyde cross-linked bovine serum albumin and its preparation and application |
| CN111420067A (en)* | 2020-03-09 | 2020-07-17 | 西南交通大学 | A kind of composite microsphere nanocarrier and its preparation method and application |
| CN111892907A (en)* | 2020-07-15 | 2020-11-06 | 广东省医疗器械研究所 | Albumin adhesive and preparation method thereof |
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