Ion Exchange Chromatography Services
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Ion exchange chromatography is a powerful technique used in the separation and purification of charged molecules, such as proteins, peptides, and nucleic acids. It is based on the principle of selectively binding charged analytes to oppositely charged stationary phases, typically a resin or gel matrix, under conditions that promote the desired interactions.
The team at GL Technologies fully understands the basics of ion exchange chromatography, including its mechanism of action, types of stationary phases, and applications in research and industry.
To speak with the experts about your Ion Exchange HPLC Calibration needs, please fill out our online form or give us a call!
Mechanism of Action
Ion exchange chromatography relies on the interaction between charged analytes and oppositely charged stationary phases. The stationary phase can be an anion exchanger, which binds positively charged analytes, or a cation exchanger, which binds negatively charged analytes.
The binding strength between the analyte and the stationary phase is determined by the charge density, size, and shape of the analyte, as well as the charge density, type, and size of the stationary phase. The pH, salt concentration, and buffer composition of the mobile phase also play important roles in modulating the binding interactions.
In general, a sample containing a mixture of charged analytes is loaded onto the ion exchange column, which contains the stationary phase. The mobile phase, typically a buffer solution, is then passed through the column to elute the bound analytes in order of decreasing strength of interaction. By adjusting the conditions of the mobile phase, such as the pH and salt concentration, the selectivity and resolution of the separation can be optimized.
Types of Stationary Phases
There are two main types of stationary phases used in ion exchange chromatography: resin-based and gel-based matrices.
Resin-based matrices are typically made of a porous polymer, such as polystyrene-divinylbenzene, that has been functionalized with charged groups, such as carboxylate or quaternary ammonium, to create the anion or cation exchanger, respectively. These resins have high binding capacity, fast kinetics, and good stability under a wide range of pH and salt conditions.
Gel-based matrices, on the other hand, are typically made of a cross-linked hydrophilic polymer, such as agarose or dextran, that has been functionalized with charged groups to create the anion or cation exchanger. These gels have lower binding capacity and slower kinetics than resin-based matrices, but they offer better resolution and are more gentle on delicate analytes, such as proteins.
Applications
Ion exchange chromatography has a wide range of applications in research and industry, including protein purification, nucleic acid isolation, and antibody purification. It is often used in combination with other chromatographic techniques, such as size exclusion chromatography and affinity chromatography, to achieve higher purity and yield.
In protein purification, ion exchange chromatography is commonly used as a second or third step after initial crude purification. It can separate different isoforms of a protein, such as those with different post-translational modifications or charges, as well as remove contaminants such as host cell proteins or DNA.
In nucleic acid isolation, ion exchange chromatography is used to separate single-stranded DNA or RNA from double-stranded DNA or RNA, as well as remove contaminants such as salts or proteins.
In antibody purification, ion exchange chromatography is used to separate different forms of an antibody, such as the intact antibody, the Fab fragment, or the Fc fragment, as well as remove contaminants such as host cell proteins or endotoxins.
To speak with the experts about your Ion Exchange HPLC Calibration needs, please fill out our online form or give us a call!
Ion exchange chromatography is a powerful technique that has revolutionized the separation and purification of charged analytes. It offers high selectivity, high resolution, and high yield, and can be used in a wide range of applications. By understanding the principles of ion exchange chromatography and the