說明
貝融生物生產的重組豬胰蛋白酶（Trypsin porcine sequence，rTP-GMP）是由基因工程菌表達并層析純化獲得。rTP-GMP的氨基酸序列與豬胰腺來源的陽離子型胰蛋白酶完全一致，可替代豬胰腺來源的胰蛋白酶應用于各種生物技術過程中。rTP利用畢赤酵母表達。貝融生物承諾生產過程不使用任何動物來源成分，包括采用植物培養(yǎng)基，并出具無TSE/BSE申明。rPT-GMP由基因工程菌生產，其氨基酸序列與豬胰腺胰酶完全一致，特異性相同。rPT-GMP相對分子質量約為24kD，最適工作pH為7-10。與動物源性胰酶相比，rPT-GMP雜蛋白少，一般通過親和層析的方式純化，產品純度極高；因為不含各種動物源性蛋白（尤其是糜蛋白酶等），酶消化效果更好，不會對消化效果產生負面影響；無外源因子污染的風險。在消化細胞的過程中，胰酶的實際使用量極大的減少，更利于胰酶殘留指標的控制，對提高最終產品的質量具有積極意義。
胰蛋白酶（Trypsin；EC 34.4.21.4，CAS：9002-07-7）是一種絲氨酸蛋白酶，酶解賴氨酸及精氨酸C末端形成的肽鍵。WHO在生物學標準化委員會第65號報告中指出（點擊查看），推薦采用重組胰酶。《 中國藥典》三部（2015）版和《歐洲藥典》8.0版也明確要求，動物源胰酶用于疫苗生產時，需檢測胰酶相關的外源性病毒，包括豬細小病毒、牛細小病毒、PCV等。因檢測方法靈敏度的限制，目前不能完全消除外源性病毒污染的風險。
在基因工程胰島素的生產中，胰蛋白酶能將胰島素原酶切為有活性的胰島素。目前胰蛋白酶主要從豬、牛的胰臟中提取，但因其動物源性，存在未知病毒和外源因子污染的風險，且常因分離純化不徹底，存在其它酶類（如糜蛋白酶等）對底物非特異性酶切的風險。另外，由于動物胰腺的個體差異，特別是胰腺中脂肪含量嚴重影響酶活力以及酶的活化和純化，故每批的活性胰蛋白酶收率存在差異。同時，傳統(tǒng)制備方法中分離純化的步驟多、時間長，易對蛋白造成破壞以致失活，導致產率偏低。因此，提取類的胰蛋白酶在胰島素等蛋白藥物生產和分析中的應用受到較大限制。
產品用途
重組胰島素生產；
質量文件完整：按客戶需求，可提供相關法規(guī)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.
質量標準
| 檢測項目 | 標準 |
外觀 | Sterile Filtered White lyophilized (freeze-dried) powder and sterile clear, colorless liquid |
氨基酸分析（賴氨酸） | 39 |
分子量 | 23KDa |
純度 | 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) |
內毒素 | ＜0.5 EU/100U as determined by TAL method |
無菌 | Negative |
支原體 | 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 |
外觀：無色澄清透明或白色狀粉末 |
分子量：23kDa |
宿主DNA殘留 ≤ 0.02 ng/μg |
宿主蛋白殘留 ≤ 0.5 ng/μg |
酶活 ≥ 3800 USP U/g |
純度：β-trypsin≥70%，α-trypsin≤20%（SDS-Page/HPLC） |
內毒素 ＜0.5 EU/100U |
無菌：陰性 |
| 支原體：陰性 |
外源病毒：陰性 |
重金屬殘留 ≤ 10 ppm |
儲存分類代碼 | WGK | 閃點（F） |
8A - Combustible, corrosive hazardous materials | WGK 3 | Not Applicable |
閃點（C) | 個人防護裝備 | |
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. |
| 產品使用 |
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. |
| 培養(yǎng)基制備 |
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. |
