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Effective method for purification of lysozyme from human urine
1Department of Internal Medicine, Branch Hospital, Nagoya University School of Medicine, Japan.
Journal of Chromatography. B, Biomedical Applications
|October 11, 1996
Summary
Researchers purified lysozyme from hemodialysis patient urine using DEAE Sephadex and Sephacryl chromatography. The purified protein confirmed as lysozyme, showing high purity and identical N-terminal sequence.
Area of Science:
- Biochemistry
- Protein Purification
- Clinical Chemistry
Background:
- Lysozyme is an enzyme with antimicrobial properties.
- Urinary lysozyme levels can be altered in patients undergoing hemodialysis.
- Efficient purification methods are crucial for studying lysozyme's role in renal disease.
Purpose of the Study:
- To purify and characterize lysozyme from the urine of a hemodialysis patient.
- To confirm the identity and purity of the isolated protein.
Main Methods:
- Two-step purification involving DEAE Sephadex and Sephacryl S-100 chromatography.
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for purity assessment.
- Molecular mass determination and N-terminal amino acid sequencing for protein identification.
Main Results:
- A two-step chromatographic procedure successfully isolated lysozyme.
- The purified protein exhibited a single band on SDS-PAGE, indicating high purity.
- The molecular mass was determined to be 14,500 Da, consistent with lysozyme.
- N-terminal amino acid sequencing confirmed the protein's identity as lysozyme.
Conclusions:
- Lysozyme can be effectively purified from the urine of hemodialysis patients.
- Sephacryl S-100 chromatography demonstrates specific affinity for lysozyme, enhancing purification efficiency.
- This study validates a method for obtaining pure lysozyme for further investigation in clinical settings.