Antibody Protein Purification

Monoclonal and polyclonal antibodies are commonly used in basic research and drug discovery. Antibody-based biologics also comprise a growing proportion of new therapeutics. Affinity chromatography using Protein A, Protein G, and Protein L crosslinked media plays a major role in the isolation and production of purified antibodies for lab scale, process scale, and industrial scale workflows.


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Agarose beads and affinity gels

Agarose beads and affinity gels enable antibody purification using Protein A or Protein G cross-linked to beaded agarose. 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. Immunoprecipitation kits include affinity gels, mini-spin columns, and lysis reagent and are specifically designed to allow maximal recovery of proteins from immunoprecipitates.

Fast Flow Sepharose™ and Fast Flow agarose resins

Fast Flow Sepharose™ and Fast flow agarose use highly crosslinked agarose beads or gels that can withstand higher pressures compared to standard agarose bead resins. This makes these resins suitable for fast protein liquid chromatography (FPLC) purification methods. These resins are recommended for medium to large scale antibody purification.

PureProteome™ Magnetic Bead System

PureProteome™ Protein A, Protein G, and Protein A/G magnetic beads provide a rapid and reproducible way to purify immunoglobulins from complex mixtures such as serum, plasma or cell culture supernatant samples. 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 automation.

Cytiva™ Amersham chromatography columns and media

A wide range of Cytiva™ Amersham chromatography media are available for purification of monoclonal and polyclonal antibodies from different sources.

  • GraviTrap™ columns are prepacked gravity-flow columns designed for fast and efficient manual purification of monoclonal and polyclonal antibodies, antibody fragments from cell culture supernatant, and biological fluids.
  • SpinTrap™ columns are prepacked, single-use spin columns for simple, small-scale purification of antibodies or enrichment of specific proteins.
  • HiTrap™ HP columns are prepacked with Protein A or Protein G Sepharose™ High Performance resin for routine, preparative purification of monoclonal and polyclonal antibodies such as human IgG. HiTrap™ MabSelect columns are prepacked with MabSelect resin, a bioprocess medium for capturing monoclonal antibodies (Mabs) from large sample volumes.
  • MultiTrap™ 96-well filter plates provide a fast, reliable format for small-scale purification of antibodies or enrichment of proteins of interest using prepacked 96-well filter plates amenable to centrifugation or vacuum filtration, manually or using automated robotic systems.
  • HiScreen™ Capto L columns are prepacked with Capto L, a bioprocess affinity chromatography resin for capture of antibodies and antibody fragments such as Fab fragments, single-chain variable fragments (scFv), and domain antibodies (Dabs). HiScreen™ Capto L columns are part of the process development platform. These columns are an excellent choice for method optimization and parameter screening.
  • HiScreen™ MabSelect™ columns are prepacked with MabSelect™ resin and are part of the process development platform available from Cytiva. MabSelect™ resin is a bioprocess resin for capture of monoclonal antibodies from large sample volumes. These columns are an excellent choice for method optimization and parameter screening for capture of monoclonal antibodies.
  • Protein A and Protein G 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 monoclonal and polyclonal antibodies from serum and cell supernatants.
  • GammaBind and GammaBind Plus Sepharose™ are Protein G affinity chromatography resins for purifying immunoglobulins (IgGs) from a variety of species, and is used to analyze and purify classes, subclasses, and fragments of immunoglobulins from any biological fluid or cell culture medium. This resin is well suited for immunoprecipitation procedures.
  • Protein A and Protein G Sepharose™, Sepharose™ Fast Flow, Capto L, and MabSelect™ resins are also available separately for scaling procedures.

Table: Immunoglobulin affinity for Protein A/Protein G/Protein L




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