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Polyelectrolyte complexes as vehicles for affinity precipitation of proteins
1Department of Biotechnology, Lund University, Sweden.
Journal of Biotechnology
|November 29, 1996
Summary
This study demonstrates effective purification of lactate dehydrogenase (LDH) using polyethylene imine (PEI) and polyacrylic acid (PA) polyelectrolyte complexes (PECs). The method achieved high yield and purity, showcasing potential for protein purification applications.
Area of Science:
- Biochemistry
- Polymer Science
- Protein Purification
Background:
- Polyelectrolyte complexes (PECs) formed from polyethylene imine (PEI) and polyacrylic acid sodium salt (PA) exhibit tunable solubility.
- Cibacron blue 3GA (CB) conjugation to PEI enables specific biomolecule interactions.
Purpose of the Study:
- To develop an affinity precipitation method for purifying lactate dehydrogenase (LDH) from beef heart extracts.
- To investigate the recovery and reusability of PECs in protein purification.
Main Methods:
- Formation of PECs using PEI and PA, with CB conjugation to PEI.
- Affinity precipitation of LDH induced by pH shift.
- Desorption and separation of LDH using KCl and pH adjustment.
- Removal of nucleic acids using PEI precipitation prior to LDH purification.
Main Results:
- LDH was purified with an 85% yield and an approximate 11-fold increase in purity.
- Recovered PECs showed similar performance upon single reuse.
- Interfering nucleic acids were effectively removed.
Conclusions:
- PEI-PA PECs functionalized with Cibacron blue offer an efficient method for LDH affinity precipitation.
- The developed method provides high yield and purity, with potential for polymer recovery and reuse.