title: Mild conditions for releasing mono and bis-biotinylated macromolecules from immobilized streptavidin.
abstract: The high affinity (kd= approximately 10(-15)M) of streptavidin and avidin for biotin is key to a large number of biological applications and is essentially irreversible unless the complex is exposed to harsh conditions (e.g. heat (100 degrees C for 10 min)), detergents, and/or denaturants which damage macromolecules. Thus, high binding affinity becomes a disadvantage when a biotinylated target must be released for further processing. This work describes relatively mild conditions that release biotin and mono- and bis-biotinylated macromolecules from immobilized streptavidin on monodispersed magnetic beads.
authors.affiliation: Molecular Biology Research Laboratory, and Departments of Biomedical Engineering, Biology and Pharmacology, 36 Cummington Street, Boston, MA 02215, USA.
authors.forename: Giang H
authors.initials: GH
authors.lastname: Nguyen
authors.forename: Jaqueline S
authors.initials: JS
authors.lastname: Milea
authors.forename: Anurag
authors.initials: A
authors.lastname: Rai
authors.forename: Cassandra L
authors.initials: CL
authors.lastname: Smith
mesh_terms: D001360:Avidin / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry; D001665:Binding Sites; D001710:Biotin / Q000737:chemistry; D019921:Biotinylation; D007700:Kinetics; D008024:Ligands; D046911:Macromolecular Substances / Q000737:chemistry*; D008280:Magnetics; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D019809:Streptavidin / Q000737:chemistry*; D013696:Temperature; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D008024:Ligands; D046911:Macromolecular Substances; D001360:Avidin; D001710:Biotin; D019809:Streptavidin