Description
Protein G HTC Agarose Beads are high-flow affinity chromatography beads designed for purification and isolation of IgG antibodies from cell culture media, biological fluids, and other antibody-containing samples.
Recombinant Protein G is covalently immobilized on highly cross-linked 4% agarose beads, providing approximately 20 mg human IgG binding capacity per mL of resin. The HTC matrix supports high flow rates and can be used for batch or column purification as well as FPLC, HPLC, and process-scale antibody purification.
Recombinant Protein G contains only the IgG-binding domains of native Protein G. The albumin-binding and cell-associated domains have been removed to maximize specific IgG binding. Protein G binds a broad range of IgG species and subclasses, including antibodies that may have lower affinity for Protein A. Binding capacity can vary depending on the source and subclass of the immunoglobulin, so antibody compatibility should be considered when selecting between Protein G and other affinity ligands.
The highly cross-linked HTC agarose matrix provides the rigidity and mechanical resistance needed for faster flow rates while maintaining good chromatographic performance. Protein G is covalently attached to the matrix to minimize ligand leakage and allow the resin to be regenerated and reused when properly handled. These properties make Protein G HTC Agarose Beads useful for antibody purification, immunoprecipitation, and applications requiring scalable or higher-flow antibody separation.
Binding Capacity
~20 mg Human IgG / ml resin
Mechanism
Protein G HTC Agarose Beads capture antibodies through the natural affinity of Protein G for immunoglobulins. Immobilized recombinant Protein G binds primarily to the Fc region of compatible IgG antibodies. This allows antibodies to remain bound to the agarose matrix while unbound proteins and other sample components are removed during washing.
Protein G has a different species and IgG subclass binding profile than Protein A. It generally provides strong binding to a broad range of mammalian IgGs, including several antibodies with weaker affinity for Protein A. Researchers should consider the species and subclass of the antibody when selecting between Protein G, Protein A, and other antibody-binding ligands.
Common Applications
(Click each for more information)
IgG Antibody Purification
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Purpose: To selectively isolate IgG antibodies from serum, biological fluids, cell culture supernatants, and other antibody-containing samples.
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How It Works: Immobilized Protein G binds primarily to the Fc region of compatible IgG molecules. Other sample components are removed during washing, and the captured antibodies are subsequently eluted from the resin.
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Applications: Human and mouse IgG purification, polyclonal antibody purification, antibody isolation from serum, and preparation of purified antibodies for downstream research.
Fishman, J. B., & Berg, E. A. (2019). Protein A and Protein G purification of antibodies. Cold Spring Harbor Protocols, 2019(1), pdb.prot099143. https://doi.org/10.1101/pdb.prot099143
Monoclonal Antibody Purification from Cell Culture
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Purpose: To recover secreted monoclonal antibodies from hybridoma or recombinant cell culture supernatants.
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How It Works: Protein G selectively captures compatible IgG from clarified culture media while media proteins and other contaminants pass through or are removed during washing.
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Applications: Purification of hybridoma-derived antibodies, recombinant monoclonal antibody purification, antibody screening, and preparation of antibodies for characterization.
Aybay, C., & Imir, T. (2000). Development of a rapid, single-step procedure using protein G affinity chromatography to deplete fetal calf serum of its IgG and to isolate murine IgG1 monoclonal antibodies from supernatants of hybridoma cells. Journal of Immunological Methods, 233(1–2), 77–81. https://doi.org/10.1016/S0022-1759(99)00195-7
High-Flow and Scalable Antibody Purification
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Purpose: To purify antibodies using chromatography systems that require higher flow rates, increased operating pressures, or larger sample volumes.
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How It Works: The highly cross-linked HTC agarose matrix provides increased mechanical strength and resistance to compression. This allows Protein G affinity capture to be performed using FPLC, HPLC, and other higher-flow chromatography systems.
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Applications: Automated antibody purification, process development, larger-volume IgG purification, and scale-up studies.
Hage, D. S., Anguizola, J. A., Bi, C., Li, R., Matsuda, R., Papastavros, E., ... & Zheng, X. (2012). Pharmaceutical and biomedical applications of affinity chromatography: Recent trends and developments. Journal of Pharmaceutical and Biomedical Analysis, 69, 93–105.
Purification of IgG with Limited Protein A Affinity
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Purpose: To isolate antibodies from species or subclasses that bind more effectively to Protein G than Protein A.
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How It Works: Protein G has a broader IgG-binding profile than Protein A for several commonly studied antibodies. This includes strong affinity for human IgG subclasses and mouse IgG, including mouse IgG1, which binds poorly to Protein A.
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Applications: Mouse monoclonal antibody purification, human IgG purification, antibody method development, and selection of an affinity resin based on species and subclass.
Björck, L., & Kronvall, G. (1984). Purification and some properties of streptococcal protein G, a novel IgG-binding reagent. Journal of Immunology, 133(2), 969–974. https://doi.org/10.4049/jimmunol.133.2.969
Key Benefits
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Broad IgG Binding: Protein G binds IgG from a wide range of mammalian species and subclasses, including several antibodies with limited affinity for Protein A.
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High-Flow HTC Matrix: Highly cross-linked agarose provides the mechanical strength needed for faster flow rates and operation at pressures up to 300 kPa.
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Suitable for FPLC and HPLC: The rigid HTC support makes the resin appropriate for automated chromatography systems in addition to conventional gravity-flow purification.
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Strong Antibody Binding Capacity: Provides approximately 20 mg human IgG binding capacity per mL of resin for efficient research-scale antibody purification.
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Stable, Reusable Affinity Matrix: Protein G is covalently immobilized on the agarose support to minimize ligand leakage, and the resin can be regenerated and reused when properly handled.
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Scalable Purification: The combination of Protein G affinity and a mechanically robust HTC matrix supports applications ranging from routine antibody isolation to larger-volume and process-development studies.
Storage/Handling
Store at 2 to 8°C.
Do not freeze
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