说明
贝融生物生产的重组猪胰蛋白酶（Trypsin porcine sequence，rTP-GMP）是由基因工程菌表达并层析纯化获得。rTP-GMP的氨基酸序列与猪胰腺来源的阳离子型胰蛋白酶完全一致，可替代猪胰腺来源的胰蛋白酶应用于各种生物技术过程中。rTP利用毕赤酵母表达。贝融生物承诺生产过程不使用任何动物来源成分，包括采用植物培养基，并出具无TSE/BSE申明。rPT-GMP由基因工程菌生产，其氨基酸序列与猪胰腺胰酶完全一致，特异性相同。rPT-GMP相对分子质量约为24kD，最适工作pH为7-10。与动物源性胰酶相比，rPT-GMP杂蛋白少，一般通过亲和层析的方式纯化，产品纯度极高；因为不含各种动物源性蛋白（尤其是糜蛋白酶等），酶消化效果更好，不会对消化效果产生负面影响；无外源因子污染的风险。在消化细胞的过程中，胰酶的实际使用量极大的减少，更利于胰酶残留指标的控制，对提高最终产品的质量具有积极意义。
胰蛋白酶（Trypsin；EC 34.4.21.4，CAS：9002-07-7）是一种丝氨酸蛋白酶，酶解赖氨酸及精氨酸C末端形成的肽键。WHO在生物学标准化委员会第65号报告中指出（点击查看），推荐采用重组胰酶。《 中国药典》三部（2015）版和《欧洲药典》8.0版也明确要求，动物源胰酶用于疫苗生产时，需检测胰酶相关的外源性病毒，包括猪细小病毒、牛细小病毒、PCV等。因检测方法灵敏度的限制，目前不能完全消除外源性病毒污染的风险。
在基因工程胰岛素的生产中，胰蛋白酶能将胰岛素原酶切为有活性的胰岛素。目前胰蛋白酶主要从猪、牛的胰脏中提取，但因其动物源性，存在未知病毒和外源因子污染的风险，且常因分离纯化不彻底，存在其它酶类（如糜蛋白酶等）对底物非特异性酶切的风险。另外，由于动物胰腺的个体差异，特别是胰腺中脂肪含量严重影响酶活力以及酶的活化和纯化，故每批的活性胰蛋白酶收率存在差异。同时，传统制备方法中分离纯化的步骤多、时间长，易对蛋白造成破坏以致失活，导致产率偏低。因此，提取类的胰蛋白酶在胰岛素等蛋白药物生产和分析中的应用受到较大限制。
产品用途
重组胰岛素生产；
质量文件完整：按客户需求，可提供相关法规支持文件。
One USP Unit will produce an A253nm of 0.003 per minute with BAEE as substrate at pH7.6 at 25℃ in a reaction volume of 3.2mL
1. Solution preparation: Melt Recombinant Trypsin at room temperature, take an appropriate amount of trypsin according to the protein content of COA, and add HBSS equilibration solution (or other buffer suitable for cell digestion. If necessary, add EDTA to a final concentration of 0-1mM, It is best not to exceed 2 mM). The recommended trypsin concentration is about 0.1-0.3mg / ml (concentration adjusted according to different cells), and gently mixed; this step can be operated at room temperature
2. Use a 0.22 μm filter to filter the trypsin solution and transfer it to a sterile container; this step can be operated at room temperature;
3. The filtered trypsin solution should be used directly as required on the day (for example, adding 1ml to T25 bottles, digesting cells at 37 °C), and the liquid can be stored at 2-8 °C for 1-2 weeks.
4. Long-term storage: If the prepared recombinant phosphatase cell digestion solution needs to be stored for a long time, it can be stored stably at -20 °C for 12 months.
20MM CaCl2，10MM HCl，pH 2±0.5
For cell culture
1. Tissue block digestion, primary cell acquisition;
2. Passage digestion of adherent cells;
3. Cell culture by microcarrier method;
4. Gently digesting stem cells;
5. Immune cell therapy, etc.
For recombinant protein
1. Recombinant insulin production;
2. Protein sequencing, peptide mapping;
3. Specific proteolytic processes such as proteomics research.
质量标准
| 检测项目 | 标准 |
外观 | Sterile Filtered White lyophilized (freeze-dried) powder and sterile clear, colorless liquid |
氨基酸分析（赖氨酸） | 39 |
分子量 | 23KDa |
纯度 | β-trypsin≥70%，α-trypsin≤20% |
活性 | ≥ 3800U/mg (One USP Unit will produce an A253nm of 0.003 per minute with BAEE as substrate at pH7.6 at 25℃ in a reaction volume of 3.2mL) |
内毒素 | ＜0.5 EU/100U as determined by TAL method |
无菌 | Negative |
支原体 | Negative |
宿主DNA | <0.02 ng/μg of protein tested by DNA Fluorescent Staining method. |
宿主蛋白 | <0.5 ng/μg of protein tested by ELISA |
外观：无色澄清透明或白色状粉末 |
分子量：23kDa |
宿主DNA残留 ≤ 0.02 ng/μg |
宿主蛋白残留 ≤ 0.5 ng/μg |
酶活 ≥ 3800 USP U/g |
纯度：β-trypsin≥70%，α-trypsin≤20%（SDS-Page/HPLC） |
内毒素 ＜0.5 EU/100U |
无菌：阴性 |
| 支原体：阴性 |
外源病毒：阴性 |
重金属残留 ≤ 10 ppm |
储存分类代码 | WGK | 闪点（F） |
8A - Combustible, corrosive hazardous materials | WGK 3 | Not Applicable |
闪点（C) | 个人防护装备 | |
Not Applicable | Eye shields, gloves | |
COMPOSITION E.Coli Recombinant Trypsin（Porcine Sequence) | |
MOLECULE WEIGHT APPROX. 23kDa | |
CAT. NO. GA04.1(liquid)/GA04.2/GA04.3/GA04.4/GA04.5 | APPEARENCE Sterile Filtered White lyophilized (freeze-dried) powder and sterile clear, colorless liquid |
ENDOTOXINES < 0.5 EU/100U | Host Cell DNA Residue <0.02 ng/μg |
PURITY β-trypsin≥70%，α-trypsin≤20% | Host Cell Protein <0.5 ng/μg |
| BIOACTIVITY ≥ 3800U/mg | |
FORMULATION 20MM CaCl2，10MM HCl，pH2±0.5 | |
USAGE For Research Use or Further Manufacture Use | |
RECONSITITUION Before use this product, please read the direction below carefully. 1. This vial must be briefly centrifuged prior to opening to bring the contents to the bottom. 2. Reconstitute in a sterile aqueous buffer to an appropriate concentration. 3. Stock solutions should be apportioned into working aliquots and stored at ≤ -20 °C. Further dilutions should be made in appropriate buffered solutions. | |
STABILITY & STORAGE 12 months from date of receipt, the product should be stored ≤-20°C. Please avoid repeated freeze-thaw cycles after reconstitution. | |
| SHIPPING The product is shipped with dry ice(liquid) or wet ice (powder). Upon receipt, store it immediately at the temperature recommended. | |
Specs
FAQs
Composition | Recombinant Trypsin (porcine sequence) |
Cat No. | GB04.1/GB04.2/GB04.3/GB04.4/GB04.5 |
| Appearence | Sterile filtered clear liquid/Sterile filtered white lyophilized (freeze-dried) powder. |
| Host | E.Coli |
| Molecule Weight | 23.29KDa |
| Biological Activity | ≥ 3800U/mg (One USP Unit will produce an A253nm of 0.003 per minute with BAEE as substrate at pH7.6 at 25℃ in a reaction volume of 3.2mL) |
| Purity | β-trypsin≥70%，α-trypsin≤20% |
| Formulation | 20MM CaCl2，10MM HCl，pH 2±0.5 |
| Usage | This GMP product can be for research use or further manufacturing use |
| Endotoxin | < 0.05 EU/100U by LAL |
| Mycoplasma | Negative |
In Vitro Virus Assay | Negative |
| Host Cell DNA | Negative |
Host Cell Protein | Negative |
| Sterility | Negative |
Reconstitution | Before use this product, please read the direction below carefully. 1. This vial must be briefly centrifuged prior to opening to bring the contents to the bottom. 2. Reconstitute in a sterile aqueous buffer to an appropriate concentration. 3. Stock solutions should be apportioned into working aliquots and stored at ≤ -20 °C. Further dilutions should be made in appropriate buffered solutions. |
| Stability&Storage | 12 months from date of receipt, the product should be stored ≤-20°C. Please avoid repeated freeze-thaw cycles after reconstitution. |
Shipping | The product is shipped with dry ice(liquid) or wet ice (powder). Upon receipt, store it immediately at the temperature recommended. |
| 产品使用 |
For cell culture 1. Tissue block digestion, primary cell acquisition; 2. Passage digestion of adherent cells; 3. Cell culture by microcarrier method; 4. Gently digesting stem cells; 5. Immune cell therapy, etc. For recombinant protein 1. Recombinant insulin production; 2. Protein sequencing, peptide mapping; 3. Specific proteolytic processes such as proteomics research. |
| 培养基制备 |
1. Melt BETAGENE™ rPT at room temperature, take an appropriate amount of rPT according to the protein content of COA, and add HBSS equilibration solution (or other buffer suitable for cell digestion. If necessary, add EDTA to a final concentration of 0-1mM. Do not exceed 2 mM). The recommended rPT concentration is about 0.1-0.3mg/mL (concentration adjusted according to different cells); 2. Filter rPT solution with 0.22 μm and transfer it to a sterile container; 3. The filtered rPT solution should be used directly as required on the day (for example, adding 1mL to T25 bottles, digesting cells at 37 °C), and the liquid can be stored at 2-8 °C for 1-2 weeks. 4. Store the digestion solution prepared at -20 °C for long term. |
