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：性能參數(shù)
介質(zhì) | QuikSep Sulfate |
基質(zhì) | 高剛性瓊脂糖 |
粒徑范圍 | 45-165μm |
平均粒徑 | 90μm |
溶菌酶載量 | ≥3mg/ml?介質(zhì) |
pH穩(wěn)定性 | 5-12（長期）?5-12（短期） |
操作壓力 | ≤0.3MPa |
流速 | （16mm× 300mm，0.1MPa）300cm/h |
貯存溶液 | 20%乙醇 |
貯存溫度 | 4-30℃ |
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表2：QuikSep?Sulfate適用性
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 |
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QuikSep Sulfate?Purification of rabies virus chromatography
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1.?The chromatography column
介質(zhì)：QuikSep Sulfate；?規(guī)格：1ml?預裝柱
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2.?Sample pretreatment
高倍濃縮后的樣品用平衡液稀釋?20?倍，上樣前?0.45um?過濾，0.45um?過濾。
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3.?buffer
A：10mM PB，pH 7.2；?B：10mM PB，1M NaCl，pH 7.2。
備注：所有溶液需?0.45um?過濾。
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4.?The purification process
流速：1ml/min?或者?30cm/h；
平衡：溶液?A?平衡?10CV（CV?指柱體積，下同）；
上樣：上樣?30CV；
清洗：溶液?A?平衡?10CV；
洗脫：溶液?B?洗脫?10CV，收集洗脫液。
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5.?Purification of map
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5.?Test results
體積 （ml） | 蛋白濃度 （ug/ml） | 抗原含量 （IU/ml） | 蛋白去除率 （%） | 收率 （%） | 濃縮倍數(shù) | 純化倍數(shù) | |
原液 | 30 | 615 | 1.51 | 94 | 75.4 | 11 | 13 |
流穿液 | 30 | 557 | 0.22 | ||||
洗脫液 | 2 | 531 | 17.08 |
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7.?conclusion
該懸浮培養(yǎng)狂犬病毒純化工藝中，病毒與?QuikSep Sulfate進行吸附結(jié)合，洗脫液中蛋白去除率≈94%，回收率≈75.4%?。
8.?Order information
產(chǎn)品 | 規(guī)格(ml) | 貨號 |
QuikSep Sulfate | 25 | HZ1010-1 |
QuikSep Sulfate | 100 | HZ1010-1 |
QuikSep Sulfate | 500 | HZ1010-1 |
QuikSep Sulfate | 1000 | HZ1010-1 |
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