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Antibody separation by hydrophobic charge induction chromatography
1BioSepra, Process division of Ciphergen Biosystems, 48 Avenue des Genottes, F-95800 Cergy Pontoise, France. egisto.boschetti@biosepra.com
Trends in Biotechnology
|July 20, 2002
Summary
Hydrophobic charge induction chromatography effectively separates antibodies from various sources. This method uses a pyridine ligand and pH-based desorption, achieving high purity without preliminary concentration.
Area of Science:
- Biochemistry
- Chemical Engineering
- Protein Chemistry
Background:
- Antibody purification is crucial for therapeutic and diagnostic applications.
- Existing methods often require harsh conditions or preliminary sample concentration.
- Developing efficient and scalable antibody separation techniques is essential.
Purpose of the Study:
- To evaluate hydrophobic charge induction chromatography (HCIC) for antibody separation.
- To investigate the use of 4-mercapto-ethyl-pyridine as a ligand in HCIC.
- To assess the compatibility of HCIC with crude biological samples.
Main Methods:
- Utilized HCIC with a 4-mercapto-ethyl-pyridine ligand.
- Performed antibody adsorption under physiological conditions.
- Achieved antibody desorption by lowering pH to induce charge repulsion.
- Analyzed antibody purity using various feedstock types.
Main Results:
- HCIC demonstrated effective antibody adsorption without prior concentration.
- Antibody desorption was successfully triggered by pH reduction.
- The method showed compatibility with crude samples regarding pH, conductivity, binding capacity, and expression levels.
- Antibody purity reached up to 98%, depending on the feedstock.
Conclusions:
- HCIC using 4-mercapto-ethyl-pyridine is a robust method for antibody purification.
- The technique offers a scalable and efficient alternative for antibody separation from diverse sources.
- Physiological adsorption and pH-controlled desorption provide a gentle yet effective purification strategy.