In the field of biotechnology, affinity chromatography is often used to purify viruses and proteins. Affinity chromatography is a kind of adsorption chromatography, in which the target substances are selectively adsorbed on the ligands of column packing, and then the buffer is changed to achieve separation and recovery. Affinity chromatography is much more accurate than gel filtration or ion exchange chromatography in recovering target components.
QuikSep Sulfate is a specific affinity filler suitable for the isolation and purification of certain viruses, virus-like particles, and certain antigens and proteins (see table 2).
Features:
1. Quick and simple (one-step purification).
2. High load.
3. Easy to enlarge.
表1：性能参数
介质 | QuikSep Sulfate |
基质 | 高刚性琼脂糖 |
粒径范围 | 45-165µm |
平均粒径 | 90µm |
溶菌酶载量 | ≥3mg/ml 介质 |
pH稳定性 | 5-12（长期） 5-12（短期） |
操作压力 | ≤0.3MPa |
流速 | （16mm× 300mm，0.1MPa）300cm/h |
贮存溶液 | 20%乙醇 |
贮存温度 | 4-30℃ |
表2：QuikSep Sulfate适用性
Viruses | Viral/Microbial Agents |
Rabies Influenza Japanese Enchephalitis Feline Leukemia Feline Herpes Feline Calicivirus Respiratory Syncytial Virus Human Herpes Simplex Human Measles Human Parainfluenza | Herpes Simplex gA and gB Glycoprotein Subunits Hepatitis B Surface Antigen Filamentous Hemagglutinin from B. pertussis Leucocytosis Promoting Factor Hemagglutinin |
QuikSep Sulfate Purification of rabies virus chromatography
1. The chromatography column
介质：QuikSep Sulfate； 规格：1ml 预装柱
2. Sample pretreatment
高倍浓缩后的样品用平衡液稀释 20 倍，上样前 0.45um 过滤，0.45um 过滤。
3. buffer
A：10mM PB，pH 7.2； B：10mM PB，1M NaCl，pH 7.2。
备注：所有溶液需 0.45um 过滤。
4. The purification process
流速：1ml/min 或者 30cm/h；
平衡：溶液 A 平衡 10CV（CV 指柱体积，下同）；
上样：上样 30CV；
清洗：溶液 A 平衡 10CV；
洗脱：溶液 B 洗脱 10CV，收集洗脱液。
5. Purification of map
5. Test results
体积 （ml） | 蛋白浓度 （ug/ml） | 抗原含量 （IU/ml） | 蛋白去除率 （%） | 收率 （%） | 浓缩倍数 | 纯化倍数 | |
原液 | 30 | 615 | 1.51 | 94 | 75.4 | 11 | 13 |
流穿液 | 30 | 557 | 0.22 | ||||
洗脱液 | 2 | 531 | 17.08 |
7. conclusion
该悬浮培养狂犬病毒纯化工艺中，病毒与 QuikSep Sulfate进行吸附结合，洗脱液中蛋白去除率≈94%，回收率≈75.4% 。
8. Order information
产品 | 规格(ml) | 货号 |
QuikSep Sulfate | 25 | HZ1010-1 |
QuikSep Sulfate | 100 | HZ1010-1 |
QuikSep Sulfate | 500 | HZ1010-1 |
QuikSep Sulfate | 1000 | HZ1010-1 |
