Hydrophobic resins have been meticulously formulated with a specialized focus on facilitating intermediate purification processes. These processes aim to eliminate host-cell contaminants from partially purified targets through the employment of hydrophobic interactions, be they robust or subtle in nature. Within the realm of protein and nucleic acid purification, there exists a selection of methacrylate-based beads expressly tailored to harness hydrophobic interactions. These beads prove to be valuable assets in purifying compounds characterized by regions featuring hydrophobic interactions of varying strengths.
Hydrophobic interaction chromatography (HIC)is a widely used method in the separation of macro-biomolecules such as proteins and peptides based on the hydrophobicity difference on the molecule surface. The biggest advantage HIC resins enjoy is their mild interaction with proteins, which helps to keep the natural structure and bioactivity of macro-biomolecules to the greatest extent.
The adsorption/separation approach (sample loading with high salinity and elution with low salinity) and chromatography principle help to make HIC resins an ideal supplementary chromatography method, as well as the adjustment technology between individual operational units. HIC resin is suitable for the separation of proteins with high hydrophobicity and excellent tolerance towards high salinity, serving as an efficient tool in the purification of macro-molecules including antibodies, vaccines, recombinant proteins, interferon, and hormones, particularly in the removal of aggregates.
How do hydrophobic interaction resins work?
Hydrophobic interaction chromatography serves as an effective method for the separation of proteins characterized by varying degrees of surface hydrophobicity. In this chromatographic process, proteins interact with a hydrophobic ligand present on the Hydrophobic Interaction Chromatography (HIC) resin within a binding buffer featuring a notably high concentration of salt.
Upon the reduction of the ionic strength of the buffer, a reversal of the hydrophobic interaction ensues. Consequently, the protein exhibiting the lowest degree of hydrophobicity is the first to be eluted. Subsequently, proteins with increasing hydrophobicity are sequentially eluted, with the most hydrophobic variant requiring a more significant reduction in salt concentration to effectively reverse the interaction and achieve elution.
What are the characteristics of hydrophobic interaction resins?
The following are the main characteristics of hydrophobic interaction resins:
Differentiated selectivity and superior separation capability
Robust stability
Unique surface bonding technology and high dynamic load
High mechanical strength and lower back pressure to improve production efficiency
Superior resolution
Flow rate independent performance
What are the applications of hydrophobic interaction resins?
Sample pretreatment of the high-end liquid phase, gas chromatography, or mass spectrometry
Capture and purification of various biomacromolecules such as antibodies, proteins, polypeptides, etc.
Removal of product-related impurities such as aggregates
Removal of product-related process contaminants such as host cell proteins
How to choose hydrophobic chromatography resin?
● Matrix and particle size
Similar to IEX resins, HIC resins are obtained by coupling hydrophobic ligands on the matrix. Bestchrom HIC resins fall into four categories, specifically, Bestarose FF(Fast Flow agarose matrix, 90 μm), Bestarose HP(High-Performance agarose matrix, 34 μm), Diamond (High rigidity agarose matrix, 90 μm) and Diamond Mustang(High Performance and high rigidity agarose matrix, 40 μm).
Thanks to their agarose-made matrix, Bestchrom HIC resins retain excellent hydrophilicity and porousness of natural polysaccharides. Among the four types of HIC resin, Diamond and Diamond Mustang enjoy better mechanical and chemical stability due to their high rigidity agarose matrix. Generally speaking, HIC resins enjoy different merits including high selectivity(due to smaller bead size) as well as tolerance towards high flow rate(due to highly rigid matrix), enabling their application over the whole process of protein chromatography from capture, intermediate purification to polishing phase.
● Levels of hydrophobicity
The key to successful HIC resin selection lies in the ligands with appropriate hydrophobicity. In other words, proteins with strong hydrophobicity require resins with weak hydrophobicity, and vice versa. The different hydrophobicity levels of common HIC ligands are as follows: Butyl-S < Butyl < Octyl < Phenyl. Phenyl ligands enjoy the strongest hydrophobicity, which can be categorized into high substitute groups(High Sub, HS) and low substitute groups(Low Sub, LS);
Octyl and Butyle are aliphatic hydrophobic ligands, among which, Octyl ligands enjoy stronger hydrophobicity than Butyl ligand due to its longer chain. Whereas, Butyl-S HIC resin possesses the weakest hydrophobicity, making it suitable for the purification of molecules with strong hydrophobicity at low salinity conditions. When exploring suitable chromatography methods, it is recommended to start with resins with the strongest hydrophobicity and make sure of the applicability of HIC resins before optimizing resin combinations.
What are the factors that affect the quality of hydrophobic interaction resin?
Resin property
Protein hydrophobicity
Buffer selection(salt type, salt concentration, pH, temperature)
Operational temperature can significantly impact HIC performance, therefore it is recommended to keep the temperature stable during the chromatography process.
Who are the major suppliers of hydrophobic interaction resins?
● GE Healthcare (Cytiva): A leading life sciences company, GE Healthcare, now known as Cytiva, offers a comprehensive range of affinity chromatography resins. Renowned for the HiTrap and Sepharose product lines, they provide solutions for protein purification and separation.
● Merck Millipore: Merck Millipore, part of Merck Group, is a global supplier of life science and analytical products. Their chromatography resins, including HisTrap and Ni Sepharose, are widely used for purifying proteins and other biomolecules.
● Bestchrom: They are China's leading chromatography resin supplier and can provide various specifications of chromatography resin. The main products include affinity resins, ion exchange resins, hydrophobic interaction resins, and other products,
● Bio-Rad: Bio-Rad is a multinational manufacturer and distributor of life science research and clinical diagnostic products. Their chromatography resin offerings, such as Profinity and CHT Ceramic Hydroxyapatite, cater to diverse purification needs.
● Thermo Fisher Scientific: Thermo Fisher Scientific is a major player in the scientific research and healthcare industry. Their chromatography resins, including Pierce and HisPur, are well-regarded for applications in protein purification and analysis.
● Agilent Technologies: Agilent Technologies is a global leader in analytical instrumentation and life sciences solutions. Their chromatography resins, like HisTrap FF, contribute to efficient protein purification processes.
● Pall Corporation (Danaher): Acquired by Danaher Corporation, Pall Corporation specializes in filtration, separation, and purification technologies. Their chromatography resins, such as His60 Ni Superflow Resin, are trusted for diverse bioseparation applications.
● Tosoh Bioscience: Tosoh Bioscience is a Japanese company known for its chromatography and separation technologies. Their chromatography resins, including Toyopearl and TSKgel, are utilized in research, bioprocessing, and pharmaceutical applications.
● Repligen: Repligen Corporation focuses on developing and commercializing bioprocessing technologies. Their chromatography resins, like CaptivA PriMAB, address the purification needs of biopharmaceutical manufacturing.
FAQs about hydrophobic interaction resins
1. What is a HIC resin?
Hydrophobic resins or HIC resins are the stationary phase of liquid chromatography. HIC resins are composed of a porous matrix, such as agarose beads coupled to a ligand. Hydrophobic resins are available in bulk or in prepacked columns for the capture and purification or polishing of biomolecules like antibodies or plasmids.
2. What are the different hydrophobic ligands?
Common ligands used during HIC include phenyl, butyl, and octyl groups. Bestchrom offers a range of hydrophobic resins that use these ligands.
3. How do I know which HIC resin to choose?
Select hydrophobic resins based on the hydrophobicity of your target biomolecules. We offer screening and selection kits to help you choose an appropriate resin for high flow, capacity, and resolution.
Conclusion
Do you need hydrophobic resins? We are a professional chromatography resin manufacturer and exporter, specializing in providing high-quality hydrophobic interaction resins to overseas customers. If you need help or services from us, please send an email or visit our website.
Reference
Mahn A. Hydrophobic interaction chromatography: fundamentals and applications in biomedical engineering[M]. INTECH Open Access Publisher, 2012.
Established in 2008, Bestchrom Biosciences Ltd. is a professional manufacturer focusing on providing high-quality chromatography resins and resin-based services and solutions.
We are a leader in resin-based separation of China with the capability to produce chromatography resins on bio-process scale. Bestchrom offers a wide range of separation resins, pre-packed columns, lab and process scales empty columns, microcarriers, and customization services.
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