Engineering streptavidin and its target ligands with both infinite binding affinity and reversible binding capability

For development of reusable biosensor chips, bioreactors, protein arrays, and matrices for affinity purification of proteins, it would be ideal for streptavidin to have extremely tight binding to its target ligands (biotin and its binding tags) and at the same time to retain the feature of reversible binding capability. To achieve this objective, a streptavidin mutein (SAVSBPM32) was engineered based on a previously engineered streptavidin variant (SAVSBPM18) that can bind both biotin and its binding tag in a reversible manner. Cysteine residues were placed in strategic positions in both the streptavidin mutein and its binding tags. Disulfide bond formation allows immobilization of tagged proteins to streptavidin. Incubation with biotin in the presence of reducing agents allows stripping off tagged proteins from streptavidin.

Name thesis:degree_name

Master of Science (MSc)

Discipline thesis:degree_discipline

Biological Sciences

Grantor dc:publisher.institution

Graduate Studies

Year dc:date.issued

2015

Author dc:creator

• Fogen, Dawson

Advisor dc:contributor.advisor

• Wong, Sui-Lam

• Biology
• Microbiology
• Biology--Molecular

Statement dc:rights

• University of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.

Language dc:language.iso

eng

DOI

http://dx.doi.org/10.11575/PRISM/25896

OAI identifier oai:identifier

oai:ucalgary.scholaris.ca:11023/2088