Ion Exchange Chromatography Services

Biopharmaceutical and related organizations rely on the GL Technologies team of experts to ensure FDA cGMP compliance, product efficacy, lot repeatability, safety of patients, utilities and manufacturing equipment repeatability, and safety of personnel. We provide full-service ion exchange chromatography and HPLC certification for our clients of all sizes throughout the state of California. The team at GL Technologies fully understands the value of ion exchange chromatography, including its mechanism of action, types of stationary phases, and applications in research and development.

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The Stationary Phase of Ion Exchange Chromatography

Ion exchange chromatography relies on the interaction between charged analytes and oppositely charged stationary phases. The stationary phase can be an an ion 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 (pH and salt concentration), the selectivity and resolution of the separation can be elevated.

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

They are 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 range of pH and salt conditions.

Gel-Based Matrices

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.

Ion Exchange Uses in Research

Ion exchange chromatography has a wide range of applications in research and development, examples include includin: protein purification, nucleic acid isolation, and antibody purification. It is often used in combination with other chromatographic techniques like exclusion chromatography and affinity chromatography, to achieve higher purity and yield.

Protein Purification

Ion exchange chromatography is a technique used in protein purification that makes use of the charge differences between proteins and their surrounding environment. Using a resin matrix containing charged groups, ions of opposite charge to the protein of interest can be immobilized and selectively bind to the resin, creating separation and purification. It a method that is highly effective for separating proteins based on their net charge and further improved by adjusting pH concentrations. This process has allowed researchers the ability to obtain biologically active proteins for research in medicine and biotech studies.

Nucleic Acid Isolation

With ion exchange chromatography and nucleic acid isolation, it relies on selectively binding charged molecules to a solid support with the help of an ion exchange resin. During this process, the nucleic acides in the sample are loaded on to the column and the resin selectively binds either positively or negatively charged nucleic acids (depending on the type of resin used). By applying specific buffer conditions, the bound nucleic acids can be eluted in a purified form, free from contaminants. This makes this technique perfect for research applications that include DNA sequencing, gene expression, and genetic engineering as a few examples.

Antibody Purification

Antibody purification relies on the principle of selectively binding charged molecules to a stationary phase with opposite charge. During this process, the mixture containing the desired antibody is loaded onto a column packed with ion exchange resin, where the target protein binds to the resin while unwanted impurities pass through. By changing the pH and ionic strength of the mobile phase, the bound antibody can be collected in a highly purified form. The antibody purification is a full asset for research in various immunological research and developmnet studies.