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Comparison of Yaxin Recombinant Aprotinin (RTI16) and Sigma Extract Aprotinin (A1153)
OBJECTIVE: The goal of this study is to compare the differences and similarities in specific activity, absorbance, acidity, and purity between Yaxin Recombinant Aprotinin (Cat. No. RTI16, Lot No. RTI161101) and Sigma Aprotinin (Cat. No. A1153, Lot No. SLBP2656V).
Experimental Method:
1. Determination of protein content, specific activity, absorbance, and acidity: A certain amount of each enzyme powder was weighed and dissolved in purified water to prepare a 2.5g/ml enzyme solution. Protein content was measured using the absorption coefficient method at 280 nm, where 1 mg/ml corresponds to an absorbance of 0.84. Activity was determined according to the standard operating procedure (SOP), and specific activity was calculated. The enzyme solution was diluted to 3 EPU/ml, and absorbance was measured at 277 nm. Acidity was tested and should not exceed 0.8.
2. Determination of HPLC purity:
- Column used: TSK-GEL G2000SWXL
- pH range: 2.5–7.5
- Flow rate: 0.5–1.0 ml/min (up to 1.2)
- Temperature: 10–30°C
- Eluent salt concentration: less than 0.5M
- Detection wavelength: 280 nm
- Method:
| Method | pH | Flow rate (ml/min) | Temperature (°C) | Eluent | Molecular sieve |
| 5.8 | 1 | 25 | 0.1M HAC-NaAC, pH 5.8, 0.1 M NaCl | TSK-GEL G2000SWXL |
- Other procedures followed the liquid chromatography SOP.
- Sample load: 20 µg
- Sample preparation: lyophilized powder dissolved in pure water.
Experimental Results:
1. Protein content, specific activity, absorbance, and acidity:
| Protein content (%) | Specific activity (EPU/mg) | Absorbance | Acidity (pH) | |
| Yaxin RTI | 72% | 3.71 | 0.505 | 5.0–7.0 |
| Sigma Aprotinin | 76.9% | 3.56 | 0.526 | 5.0–7.0 |
2. HPLC Purity:
- Yaxin Recombinant Aprotinin (RTI161101): Images show a more concentrated peak compared to Sigma Aprotinin.
- Sigma Aprotinin (A1153): Images show a broader peak.
- Both samples showed 100% purity based on HPLC analysis.
Conclusion: The results indicate that both Yaxin Recombinant Aprotinin (RTI16) and Sigma Aprotinin (A1153) have similar protein content, absorbance, and acidity. However, the specific activity of Yaxin's recombinant aprotinin was slightly higher than that of Sigma's extract. In terms of HPLC purity, both samples were found to be 100% pure, but Yaxin’s sample showed a more defined peak, suggesting better consistency and quality.
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