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Chromatography technology plays a critical role in the biopharmaceutical sector, by offering an efficient approach to achieve the separation of complex components. Chromatography resin consists of ligands and a base matrix, which serves as a critical factor in the downstream processing by determining the productivity and quality of pharmaceutical ingredients purification.
Chromatography resins can be categorized into affinity resin, ion exchange (IEX) chromatography resin, hydrophobic interaction chromatography (HIC) resin, Size exclusion chromatography (SEC) resin, and Mix-mode resin in terms of separation mode.
From this article you will learn:
Chromatography resins are a class of key materials used in chromatography processes and are widely used in the pharmaceutical, biotechnology and chemical fields. These resins are often highly selective and can effectively separate compounds in mixtures.
The choice of chromatography resin depends on the desired separation properties, including molecular size, charge, hydrophilicity and hydrophobicity, etc. Common chromatography resin types include ion exchange resins, hydrophilic and hydrophobic chromatography resins, and metal chelate chromatography resins.
These resins play a key role in the preparation and purification of biomolecules, drugs and compounds, improving the efficiency and effectiveness of chromatography processes in laboratories and industry.
There are different types of chromatography resins, and each resin has its own advantages and functions according to user requirements.
| Category | Feature |
|---|---|
| Affinity chromatography | This technique is commonly employed for the purification of proteins, antibodies, and enzymes. |
| Ion exchange (IEX) chromatography | They facilitate the exchange of ions between the resin and the sample. |
| Hydrophobic interaction chromatography | HIC resins separate biomolecules based on hydrophobic interactions |
| Size exclusion chromatography (SEC) | SEC resins separate molecules based on their size |
| Mixed-mode chromatography | They offer enhanced selectivity and flexibility |
Affinity chromatography is a chromatography method separating biomolecules based on the specific interaction among biomolecules. Thanks to its high selectivity, affinity resin is able to capture protein from complexes at a purity higher than 90% via one-step purification.
Affinity resin is widely applicable in the efficient purification of antibodies, tag proteins, and other bio-molecules with specific adsorption.
Advantages: High selectivity, excellent purity outcomes, and suitability for purifying specific target molecules.
Ion exchange (IEX) chromatography is a separation method based on the different mass and quantity of electric charges on biomolecules. Enjoying advantages including high selectivity, high binding capacity, high yield, and convenient operation, the versatile chromatography method can be used in the initial capture step as well as intermediate purification and polishing.
Therefore, it is widely applicable in the purification of electric-charged bio-molecules including amino acids, peptides, antibodies, proteins, saccharides, viruses, and nucleotides.
Advantages: High specificity for charged molecules, effective separation of biomolecules, and applicability in both analytical and preparative chromatography.
Hydrophobic interaction chromatography (HIC) is a widely used method in the separation of macro-biomolecules based on the hydrophobicity difference on the molecule surface. The biggest merit lies in its mild interaction with proteins, which enables the maintenance of natural structure and bio-activity in macro-biomolecules.
HIC adopts an adsorption mode in terms of high salinity sample loading and low salinity elution, which makes it an ideal purification option after eluting with high salinity. HIC provides an efficient method for the separation of macro-molecules including antibodies, recombinant proteins, vaccines, peptides, and nucleic acid.
Advantages: Gentle separation conditions, effective purification of proteins, and compatibility with proteins that denature in other chromatographic methods.
Size exclusion chromatography (SEC), also known as gel filtration, is a non-adsorption chromatography method. It achieves chromatography separation based on the size and shape of biomolecules, which means the key to SEC resin selection lies in the right fractionation range.
SEC enjoys the advantage of easy operation, which enables the completion of chromatography separation via a single buffer. Therefore, it is an ideal option in the desalting, buffer exchange, and polishing step of chromatography.
Advantages: Non-destructive separation method, compatibility with a wide range of biomolecules, and suitability for determining molecular weight.
Mixed-mode chromatography is an innovative chromatography method that can simultaneously provide different interactions for the binding between ligands and macro-biomolecules. Dominant interactions in this category include electric charge interaction and hydrophobic interaction.
Mixed-mode chromatography enjoys wider binding condition and a bigger operation room, which simplifies process step and boost productivity. It is widely applicable in the purification of viruses, antibodies, peptides, recombinant proteins, and nucleic acids, especially in providing effective solutions to challenges faced in the purification process.
Advantages: Versatility in separating complex samples, increased selectivity, and the ability to handle challenging separation tasks.
1. Affinity Chromatography:
Use When: High specificity is required for the separation of target molecules based on specific binding interactions.
Ideal for: Purification of proteins, antibodies, and enzymes where a strong affinity between the target and ligand is present.
2. Ion Exchange (IEX) Chromatography:
Use When: Separation based on charge differences is needed, making it suitable for molecules with varying ionization states.
Ideal for: Purification of proteins, peptides, and nucleic acids by exploiting differences in charge.
3. Hydrophobic Interaction Chromatography:
Use When: Separation is desired based on hydrophobicity, offering a milder condition compared to other hydrophobic methods.
Ideal for: Purifying proteins that are sensitive to high salt concentrations or denaturation.
4. Size Exclusion Chromatography (SEC):
Use When: Separation based on molecular size is crucial, allowing for the analysis of size distributions and purification of biomolecules.
Ideal for: Determining molecular weights and separating biomolecules based on their size without denaturation.
5. Mixed-Mode Chromatography:
Use When: Enhanced selectivity is needed by combining different separation mechanisms like ion exchange, hydrophobic interaction, and affinity.
Ideal for: Complex samples requiring versatile approaches, providing increased selectivity in bioseparation processes.
Chromatography resin plays a critical role in chromatography purification. Different combinations of functional groups and base matrices provide various functions to resins, which enables the efficient purification of complex components via a selection of the right resin.
When choosing suitable resins, the following factors should be taken into account: matrix property (polymer-based matrix provides a high flow rate due to its good mechanical property; Agarose-based resin enjoys better non-specific adsorption due to its excellent hydrophilicity), bead size(finer beads provide high selectivity while big beads can endure high flow rate), resin binding capacity (which depends on ligand types and ligand concentration) as well as scalability.
In addition, evaluation results in terms of sample yield and quality should also be considered when making the resin purchase decision.
Bestchrom Biosciences focuses on the process development and product R&D required in bio-pharmaceutical downstream processing. We take our core strength in product quality and production capacity, which enable us to continuously provide chromatography resins with more variety, better performance, and cost-effectiveness.
Contact us today by sending an email or visit our resin product page!
References
Hydrophobic interaction resins
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