胰蛋白酶是一种丝氨酸蛋白酶，能选择地水解蛋白质中由赖氨酸或精氨酸的羧基末端所构成的肽链。本品是从重组毕赤酵母中分离纯化，再经冷冻干燥得到的重组胰蛋白酶粉末。本品属于生物化学级别的基因工程酶，不含DFP、PMSF和TLCK等酶抑制剂，具有与天然胰蛋白酶相同的活性和特异性。本品活性受丝氨酸蛋白酶抑制剂如PMSF和TLCK等、金属离子螯合剂如EDTA等的抑制。
产品优势
1、无动物源性：无动物病毒、无致病物质、无外源因子污染，安全性高。
2、纯度高：比活高；宿主蛋白残留小于生物制品限度要求。
3、符合中国药典2020版和USP43的标准。
4、合规性：生产设备和生产环境符合相关法规要求，符合GMP指导原则。
产品特性
性状：白色或类白色冻干粉
分子量：23.5kDa±2.4kDa
纯度（HPLC）：α-胰蛋白酶不得大于20%; β-胰蛋白酶不得小于70%
蛋白含量：不低于40%
外源DNA残留量：≤100ng/mg pro.
菌体蛋白残留量：≤100ng/mgpro.
最佳：pH7.0~9.0
溶解性：在水或缓冲液中可溶
推荐使用方法
1、蛋白酶切使用方法
(1) 蛋白质的准备——将蛋白质溶解在25mMTris-HCl，pH8.0的溶液中。
(2) 酶液准备——称取所需的酶粉，用1mMHCl溶解，使酶液浓度为1-10mg/ml。
(3) 反应条件——酶量：目的蛋白（质量比）=1:50~1:1000，最适pH为7.0~9.0。
2、细胞消化
(1) 溶液配制：重组胰蛋白酶室温下融化，取适量胰蛋白酶，加入HBSS平衡液（或其他适宜细胞消化的缓冲液，如需要加入EDTA其终浓度推荐0-1mM，最好不超过2mM），推荐一般使用的胰蛋白酶浓度约为0.1-0.3mg/ml（浓度根据不同细胞进行调整），轻柔混匀；该步骤可在室温环境操作；
(2) 使用0.22μm滤膜对上述胰蛋白酶溶液过滤并转移至无菌容器中；该步骤可在室温环境操作；
(3) 过滤后的胰蛋白酶溶液应在当天按照要求直接使用（例如加入1ml至T25瓶，37°C消化细胞）
产品用途
1、用于生物药物生产，如胰岛素、GLP-1药物。
2、蛋白质结构与功能研究，如蛋白质质谱、测序、肽图谱分析。
储存稳定性
冻干粉在-15°C及以下条件下至少能稳定存放24个月。1mM盐酸或50mM醋酸溶解后储存于-15°C及以下，反复冻融10次，无活性损失。溶解后2~8°C保存建议不超过7天，溶解后-15°C及以下保存建议不超过30天。
注意事项与免责声明
1、本品具有吸湿性，应室温平衡后再取用。
2、本品不得用于临床诊断和治疗。仅供科研生产使用，不能用于人体。
Trypsin is a serine‑protease that selectively hydrolyzes peptide bonds formed at the carboxyl‑terminal side of lysine or arginine residues in proteins. This product is recombinant trypsin powder obtained by isolation and purification from recombinant Pichia pastoris, followed by lyophilization. It is a biochemical‑grade genetically‑engineered enzyme, free of enzyme inhibitors including DFP, PMSF and TLCK. It exhibits identical activity and specificity as native trypsin. Its activity can be inhibited by serine‑protease inhibitors (e.g. PMSF, TLCK) and metal‑ion chelators (e.g. EDTA).
Product Advantages
1. Animal‑origin‑free: Free of animal viruses, pathogenic agents and exogenous‑factor contamination for high safety.
2. High purity: High specific activity; host‑cell protein residues are below the limits specified for biological products.
3. Complies with the requirements of Chinese Pharmacopoeia 2020 Edition and USP 43.
4. Regulatory compliance: Manufacturing equipment and environment meet relevant regulatory requirements and GMP guidelines.
Product Properties
Appearance: White or off‑white lyophilized powder
Molecular Weight: 23.5 kDa ± 2.4 kDa
Purity (HPLC): α‑trypsin ≤ 20%; β‑trypsin ≥ 70%
Protein Content: ≥ 40%
Residual Exogenous DNA: ≤ 100 ng/mg pro.
Host‑Cell Protein Residue: ≤ 100 ng/mg pro.
Optimal pH: 7.0‑9.0
Solubility: Soluble in water or buffer solutions.
Recommended Protocols
1. Proteolytic Digestion
(1) Protein preparation: Dissolve target protein in 25 mM Tris‑HCl, pH 8.0.
(2) Enzyme preparation: Weigh required enzyme powder and dissolve with 1 mM HCl to prepare enzyme solution at 1‑10 mg/mL.
(3) Reaction conditions: Enzyme : target protein (mass ratio) = 1:50 ~ 1:1000; optimal pH 7.0‑9.0.
2. Cell Dissociation
(1) Solution preparation: Thaw recombinant trypsin at room temperature. Take an appropriate amount and dilute with HBSS buffer (or other suitable cell‑dissociation buffers). If EDTA is required, the recommended final concentration is 0‑1 mM and should preferably not exceed 2 mM. The typical working concentration of trypsin is 0.1‑0.3 mg/mL (adjust concentration according to cell type). This step can be performed at room temperature.
(2) Sterile‑filter the prepared trypsin solution through a 0.22 μm membrane and transfer into a sterile container. This step can be performed at room temperature.
(3) Use the filtered trypsin solution on the same day of preparation (e.g. add 1 mL into a T25 flask for cell dissociation at 37 °C).
Applications
1. Biopharmaceutical manufacturing, e.g. production of insulin and GLP‑1‑related drugs.
2. Protein structure‑function research, including protein mass spectrometry, protein sequencing and peptide‑mapping analysis.
Storage & Stability
Lyophilized powder is stable for at least 24 months stored at ≤ −15 °C. When dissolved in 1 mM HCl or 50 mM acetic acid and stored at ≤ −15 °C, no activity loss is observed after 10 freeze‑thaw cycles. After reconstitution, storage at 2‑8 °C is recommended for no more than 7 days; storage at ≤ −15 °C is recommended for no more than 30 days.
Precautions & Disclaimer
1. This product is hygroscopic. Equilibrate to room temperature before opening the vial for sampling.
2. Not for clinical diagnosis or therapeutic use. For research and manufacturing purposes only; not for human administration.
ZJ26F0333129
危险品化学品经营许可证（带存储）