Protein Chromatography
Chromatography is a technique for separating components in a mixture. Typically, the mixture is dissolved in a mobile phase and flows through a stationary phase. Different components flow out of the stationary phase at different times to be separated from one another. Following the expression of recombinant proteins, target proteins usually co-exist with host cell proteins (HCPs) and other contaminants such as nucleic acids and lipids in host cells. HCPs and other contaminants affect the quality of target protein in medical applications as they are very likely to cause inflammations, toxicity, and other side effects. Fast protein liquid chromatography (FPLC) is an extensively adopted tool to remove impurities from recombinant proteins and prepare target proteins. Different types of FPLC are grounded in distinct separation mechanisms, thus can achieve the removal of almost all types of impurities. Chromatography is usually the core step in the purification pathway. Another advantage of FPLC is its high scalability, which makes mass production of purified proteins convenient and inexpensive.
Creative BioMart Nanocage develops fast and cost-saving purification chromatography-based pathways to obtain high-purity recombinant protein nanocages for our customers. Our other purification techniques such as protein precipitation, high-performance liquid chromatography, and ultrafiltration are usually used in combination with chromatography to achieve a better, faster, and more affordable purification process.
What We Offer
Creative BioMart Nanocage owns a wide range of columns and resins for FPLC manufactured by Cytiva and Thermo Fisher. Our chromatography column volume ranges from 1 mL to over 1 L so as to conduct purification service at various scales. We also have 96-well plate resins for high throughput screening.
Fig 1. Different Scales of Chromatography Service We offer
The chromatography types and resins we include are listed in the table below.
Table 1. Chromatography for Recombinant Protein Purification
| Types of Chromatography | Seperation Mechanism | Description |
|---|---|---|
| Ion-exchange Chromatography (IEC) | Charge density differences between the protein of interest and other impurities. | Strong and weak cation/anion IEC columns are provided, including but not limited to the classical Q Fast Flow, DEME Fast Flow, SP Fast Flow, and CM Fast Flow. Columns with different resin bead sizes are available for different resolution requirements. High-resolution Mono series columns (Mono Q , Mono S) suitable for separating target proteins from similar impurities are available. Columns with porous resin beads such as Poros HQ, Poros HS are good choices in separating large proteins such as protein nanocages because of the easier diffusion of target protein into resin beads. |
| Hydrophobic interaction Chromatography (HIC) | Hydrophobic interaction differences between the protein of interest and other impurities. | HIC columns differ in binding strength, functional group density, and resin bead sizes are all used in our HIC service. Commonly used columns with Butyl-S, Butyl, Octyl, Phenyl functional groups are available. |
| Affinity Chromatography (AC) | Specific affinity of the protein of interest to the functional group on stationary phase. | AC columns include metal ion affinity column, glutathione columns, Protein A column for antibody purification, Heparin column for DNA-binding protein purification, and so on. |
| Size-exclusion Chromatography (SEC) | Molecular hydrodynamic size differences between the protein of interest and other impurities. | Resins of SEC column, in theory, do not interact with sample components. Creative BioMart Nanocage provides SEC columns for separating proteins in different molecular weight ranges. Our SEC column include Sephadex 75, Sephadex 200, Superose 6 increase, Sephacryl S-100, S-200, S-300, S-400, S-500, etc. |
| Mutimodal Chromatography (MC) | A combinantion of SEC with IEC or SEC with HIC. | Columns such as Capto core and Capto adhere series are suitable for proteins that are challenging for other types of chromatography. |
| Desalting and Buffer Exchange | Molecular hydrodynamix size difference between proteins and buffer salts | We own G10 and G25 desalting columns in different column volumes. |
Our Serive Advantages
Creative BioMart Nanocage is equipped with a variety of advanced devices for protein chromatography, including AKTA Start, AKTA Pure, AKTA Avant, AKTA PCC, AKTA Pilot, and AKTA Process. Our service is designed to be comprehensive and customized to meet distinct requirements from our customers. Final product purity can be > 85%, > 90%, > 95%, and > 99%. Product mass ranges from milligram to kilogram level. Our chromatography team has long-proven experiences in recombinant protein purification. We cover the purification of recombinant protein nanocages, engineered protein nanocages, and other recombinant proteins of our customer's interest.
If you are interested in our protein chromatography service, please do not hesitate to contact us through online inquiry.
