alexa Protein interaction with immobilized metal ion affinity ligands under high ionic strength conditions.
Chemical Engineering

Chemical Engineering

Journal of Bioprocessing & Biotechniques

Author(s): Jiang W, Hearn MT, Jiang W, Hearn MT

Abstract Share this page

Abstract An investigation into the adsorption behavior under batch equilibrium binding conditions of hen egg white lysozyme (HEWL) with the immobilized metal ion affinity chromatographic adsorbent, Cu(2+)-IDA Sepharose CL-4B, in the presence of high concentrations of NaCl or KCl has been undertaken. When the concentration of NaCl or KCl in the adsorption buffer was < or = 0.2 M, the adsorption data correlated with predictions based on the Langmuir model. However, when the salt concentration was > or = 0.5 M, the adsorption data appeared to follow the Freundlich-Langmuir model under these higher ionic strength conditions. Under elevated ionic strength conditions, the protein-ligand interactions were shown to involve positive cooperativity by Scatchard plot analysis of the experimental data. The number, n, of interacting protein molecules increased with the salt concentration, while the apparent dissociation constant Kd under these high ionic strength conditions was less dependent on the type of salt present in the adsorption buffer. These studies reveal that the binding of Cu(2+)-IDA Sepharose CL-4B to HEWL acceptor molecules under conditions of high ionic strength is characteristic of general heterogeneous interactions, whereby the immobilized metal ion affinity chromatography (IMAC) ligands bind to a protein acceptor undergoing isodesmic indefinite self-association and multisite attachment. As a consequence, these results on the adsorption behavior of HEWL have important general implications for the preparative use of immobilized metal ion affinity adsorbents in packed and expanded bed systems, particularly with regard to the resolution and productivity of the separation for the target protein. In addition, the results from these studies are relevant to investigations involving analytical affinity chromatographic or biosensor assessment of the interaction of proteins with immobilized IMAC ligands when high ionic strength equilibration feedstocks or conditions are employed. This article was published in Anal Biochem and referenced in Journal of Bioprocessing & Biotechniques

Relevant Expert PPTs

Relevant Speaker PPTs

Recommended Conferences

  • 17th Euro Biotechnology Congress
    September 25-27, 2017 Berlin, Germany
  • 2nd World Biotechnology Congress
    December 04-06, 2017 Sao Paulo, Brazil

Relevant Topics

Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2017-18
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

Agri, Food, Aqua and Veterinary Science Journals

Dr. Krish

[email protected]

1-702-714-7001 Extn: 9040

Clinical and Biochemistry Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Business & Management Journals

Ronald

[email protected]

1-702-714-7001Extn: 9042

Chemical Engineering and Chemistry Journals

Gabriel Shaw

[email protected]

1-702-714-7001 Extn: 9040

Earth & Environmental Sciences

Katie Wilson

[email protected]

1-702-714-7001Extn: 9042

Engineering Journals

James Franklin

[email protected]

1-702-714-7001Extn: 9042

General Science and Health care Journals

Andrea Jason

[email protected]

1-702-714-7001Extn: 9043

Genetics and Molecular Biology Journals

Anna Melissa

[email protected]

1-702-714-7001 Extn: 9006

Immunology & Microbiology Journals

David Gorantl

[email protected]

1-702-714-7001Extn: 9014

Informatics Journals

Stephanie Skinner

[email protected]

1-702-714-7001Extn: 9039

Material Sciences Journals

Rachle Green

[email protected]

1-702-714-7001Extn: 9039

Mathematics and Physics Journals

Jim Willison

[email protected]

1-702-714-7001 Extn: 9042

Medical Journals

Nimmi Anna

[email protected]

1-702-714-7001 Extn: 9038

Neuroscience & Psychology Journals

Nathan T

[email protected]

1-702-714-7001Extn: 9041

Pharmaceutical Sciences Journals

John Behannon

[email protected]

1-702-714-7001Extn: 9007

Social & Political Science Journals

Steve Harry

[email protected]

1-702-714-7001 Extn: 9042

 
© 2008-2017 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version
adwords