Biotinylated Protein Purification

Biotinylated proteins and biotinylated antibodies are commonly used in basic research and drug discovery. Various affinity chromatography resins and prepacked columns and plates are available to bind, isolate, and purify biotinylated proteins and antibodies.

• Agarose Beads and Affinity Gels
• Magnetic Beads
• SigmaScreen™ Streptavidin-coated High Capacity Plates
• Cytiva™ Amersham Chromatography Columns and Media

Agarose beads and affinity gels

Agarose beads and affinity gels enable biotinylated protein and biotinylated antibody purification using streptavidin or avidin cross-linked to beaded agarose. Both avidin and streptavidin bind to biotin with high affinity, though avidin can exhibit high non-specific binding. Extravidin®-agarose uses a modified avidin with very high affinity and specificity for biotin. Good for small to large scale purification, these resins are suitable for gravity flow column purification, low-speed centrifugation in batch processing, and low-pressure chromatography procedures. EZview™ affinity gels incorporate a red dye that allows the user to easily differentiate pellet from supernatant, resulting in less protein loss during batch purification.

Magnetic beads

PureProteome™ Streptavidin magnetic beads provide a rapid and reproducible way to purify immunoglobulins from complex mixtures such as serum, plasma or cell culture supernatant samples. Biotinylated proteins and biotinylated antibodies are purified using a magnetic rack or platform to separate beads from wash and elution fractions. This procedure is suitable for low throughput using single microfuge tubes or higher throughput using 96-well plates with or without automation. Streptavidin Magnetic Particles can also be used for purification of biotinylated proteins and biotinylated antibodies.

SigmaScreen™ Streptavidin High Capacity Coated Plates

SigmaScreen™ streptavidin-coated high capacity plates utilize a proprietary coating technology that provides substantially greater biotin binding capacity than standard streptavidin or ExtrAvidin coated plates. With binding capacity >300 pmol/well (96-well format) or >150 pmol/well (384-well format), these plates allow the captured protein to be eluted for post-capture analysis by various methods such as MALDI, ICAT or SDS-PAGE. SigmaScreen™ streptavidin-coated high capacity plates are manufactured under ISO 9002 in our GMP facility and deliver high well-to-well and plate-to-plate consistency.

Cytiva™ Amersham chromatography columns and media

A variety of Cytiva™ Amersham chromatography columns and media resins are available for reliable, high resolution small- to large-scale purification of biotinylated proteins and antibodies.

• SpinTrap™ columns are prepacked, single-use spin columns for simple, small-scale purification of biotinylated proteins and biotinylated antibodies.
• HiTrap™ HP columns are prepacked with Streptavidin Sepharose™ High Performance resin for routine, preparative purification of biotinylated proteins and antibodies.
• MultiTrap™ 96-well filter plates provide a fast, reliable format for small-scale purification of biotinylated proteins and antibodies using prepacked 96-well filter plates amenable to centrifugation or vacuum filtration, manually or using automated robotic systems.
• Streptavidin Mag Sepharose™ media are comprised of Sepharose™-based magnetic beads designed to simplify enrichment of target proteins by immunoprecipitation or pull-down applications, and can be used for rapid, small-scale purification and screening of biotinylated proteins and biotinylated antibodies from cell supernatants.
• Streptavidin Sepharose™ High Performance resin is also available separately for scaling purification.

Related Product Resources

Brochure: Refine Protein Preparation
Purification or Removal of Biotin and Biotinylated Substances