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Antibody Purification
Antibody purification involves isolation of antibody from serum (polyclonal antibody), ascites fluid, or from the culture supernatant of a hybridoma cell line (monoclonal antibody). Purification methods range from very crude to highly specific. The level of purification necessary to obtain usable antibody depends upon the intended application(s) for the antibody. Antibodies may be purified by class (e.g., IgG) without regard to antigen specificity, or affinity purified to an immobilized antigen that is antibody specific.
We offer a wide selection of ligands and formats to purify total or specific antibodies from serum, ascites fluid, or tissue culture supernatant. Our portfolio is designed to meet small-scale (screening) to large-scale (bioprocess) needs.
- Broad product selection—affinity supports for the purification and enrichment of multiple antibodies, including Protein A, Alkaline Stable Protein A, Protein G, Protein A/G, and Protein L
- More formats—magnetic beads, loose resin, spin columns and kits, FPLC cartridges, and 96-well filter plates enable antibody purification from microgram to kilogram scales
- High performance—resins are designed to maximize protein yield and reduce background
- Economical—pricing is similar to or better than our leading competitors
Choose the right antibody purification resin
| Protein A | Protein A, Alkaline Stable | Protein G | Protein A/G | Protein L | Melon Gel | |
|---|---|---|---|---|---|---|
| Target | IgG enrichment | IgG enrichment | IgG enrichment | IgG enrichment | IgG enrichment | Negative selection |
| Use for | Polyclonal IgG from rabbit, pig, dog, cat serum | Polyclonal IgG from rabbit, pig, dog, cat serum Alkaline stability characteristics under clean-in-place conditions For purification of mAbs, subpopulation of scFv’s and Fab fragments containing a VH3 domain | IgG from mouse, human, cow, goat and sheep | Polyclonal IgG from many species | Human or mouse monoclonal antibodies known to have appropriate kappa light chains | Antibodies that do not purify well using Protein A or Protein G |
| Formats | Magnetic beads, filter plates, loose resins (agarose and POROS), spin columns and kits, FPLC cartridges | Loose resins | Magnetic beads, filter plates, loose resins (agarose and POROS), spin columns and kits, FPLC cartridges | Magnetic beads, filter plates, loose resins (agarose and POROS), spin columns and kits, FPLC cartridges | Magnetic beads, filter plates, loose resins, spin columns and kits, FPLC cartridges | Serum and monoclonal antibody kits, FPLC cartridges, spin plate |
| Processing scale | µg–kg | µg–kg | µg–kg | µg–kg | µg–mg | µg–g |
Featured antibody purification categories
Protein A binds to human, mouse, goat, and rabbit IgG (except for human IgG3 and mouse IgG1) at the Fc portion of the intact Ig and is typically the ligand of choice when purifying IgG from rabbit polyclonal antisera.
Protein G binds all human, goat, and murine IgG and subclasses at the Fc portion of the intact Ig and is typically the ligand of choice for purifying IgG from goat or mouse polyclonal antisera.
Protein A/G is a genetically engineered protein with combined IgG binding profiles of both Protein A and G. It contains four Fc binding domains from Protein A and two from Protein G, providing additive class-specific binding properties for Protein A and G.
Protein L binds specifically to Ig kappa light chains, and members of all antibody classes (IgG, IgM, IgA, IgD, and IgE). It is the affinity molecule of choice for the positive selection of kappa Fab fragments or recombinant single-chain variable fragments.
Antigen-specific antibody purification
Choose from a variety of activated affinity supports (porous resins or magnetic beads) to successfully immobilize peptides, proteins, and other antigens, like amino, sulfhydryl, carboxyl, or glycosyl functional groups.
When you need high performance and high throughput on a large scale, POROS bulk chromatography resins are the industry standard for process-scale bioseparations.
Dynabeads and other magnetic bead and resin products and technologies for IP, co-IP, pull-down, ChIP, and other small-scale affinity purification and analysis assays.
General characteristics of Ig-binding proteins
| Native Protein A | Recombinant Protein A | Recombinant Protein G | Recombinant Protein A/G | Recombinant Protein L | |
|---|---|---|---|---|---|
| Native source | Staphylococcus aureus | Staphylococcus aureus | Groups C and G Streptococci | N/A | Peptostrepto-coccus magnus |
| Production source | S. aureus | E. coli | E. coli | E. coli | E. coli |
| Molecular mass | 46,700 | 44,600 | 21,600 | 50,460 | 35,800 |
| Apparent mass by SDS-PAGE | 42 kDa | 45 kDa | 32 kDa | 40 to 45 kDa | 36 kDa |
| Ig-binding sites (#) | 5 | 5 | 2 | 4+2 | 4 |
| Albumin binding site | No | No | No | No | No |
| Optimal binding pH | 8.2 | 8.2 | 5 | 5 to 8.2 | 7.5 |
| Ig binding target | Fc | Fc | Fc | Fc | VL-kappa |
Partner with us to scale-up your process or product development pipeline with bulk quantities of high-quality immunoassay reagents and biochemicals. We offer a variety of compounds, devices, and resins for your large-scale applications.
Resources
For Research Use Only. Not for use in diagnostic procedures.