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Achieve ultrahigh resolution and high efficiency separations of proteins and glycoproteins with a pI 3–10 and molecular weight over 10,000 units. Thermo Scientific ProPac Protein LC columns have a long-standing reputation for providing excellent reproducibility and durability under a broad range of pH, temperature, and mobile phase compositions.
ProPac columns along with Thermo Scientific MAbPac and Acclaim SEC 300 columns comprise our broader portfolio that address the challenges in biopharmaceutical separations.
Separations are based on the accessible surface charge of the protein or glycoprotein with the column’s functional group. Unsure if you need an anion or cation exchange column? The protein’s pI can help to determine which column to use.
Recommended Ion Exchange Columns by Type | |||||
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Column Type | Strong anion exchange (SAX) columns | Weak anion exchange (WAX) columns | Weak cation exchange (WCX) columns | Strong cation exchange (SCX) columns | |
Column | ProPac SAX-10 | ProPac WAX-10 | ProPac Elite WCX 5μm | ProPac WCX-10 | MAbPac SCX |
Functional group | Quaternary ammonium | Tertiary amine | Carboxylic acid | Sulfonic acid | |
Sample pI | Typically for protein pI is < 6 | Typically for protein pI is > 6 | |||
Notes | Based on WCX-10 column Ultrahigh resolution Faster, more efficient, highly reproducible | Long standing gold-standard column | Strong Cation Exchange columns offer an alternate selectivity to Weak Cation Exchange columns | ||
Gradient | Use salt gradient | WAX will buffer your phase more than SAX | Use salt or pH gradients* | Use salt gradient | Use salt or pH gradients |
* Using our CX-1 pH Gradient Buffers
With many opportunities for post translational modifications (PTMs), it is often necessary to employ multiple column supports to detect, characterize, and quantify all structural variants and modifications and to monitor product stability.
HIC columns separate proteins and mAb molecules in order of increasing hydrophobicity under non-denaturing conditions. Analytes bind to the weakly hydrophobic stationary phase in the presence of high salt concentration and elute off the column as the salt concentration decreases.
In contrast to reversed phase liquid chromatography, HIC typically preserves the biological activity of the protein, which is useful for downstream functional analysis such as binding and cell-based potency assays.
Also see our Thermo Scientific MAbPac HIC columns
MAbPac HIC-10 LC Column | MAbPac HIC-20 LC Columns | MAbPac HIC-Butyl LC Columns |
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The biotechnology and biopharmaceutical industry has an extensive development pipeline for protein and monoclonal antibody (mAb) therapeutics. We understand that mAbs have very complex structures, with many possible site-specific variations that can change safety, efficacy, and stability profiles. With such potential for post translational modifications (PTMs), it is essential to detect, characterize, and quantify structural variants and modifications and to monitor product stability.
We see it as our job to develop and deliver innovative and efficient analytic tools during mAb development and production to prove safety, efficacy, and stability to regulatory agencies. Look to Thermo Scientific HPLC and LC/MS instruments and HPLC columns as your essential tools for the characterization of mAb and other protein biotherapeutics. They have been designed to address your challenges.
Analysis | Description | Columns & Buffers |
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Charge variant analysis | Routine charge variant profiling/screening, including lysine truncation, deamidation & acylation | |
Methionine & tryptophan oxidation | Targeted analysis of methionine and tryptophan oxidation | |
Antibody drug conjugate (ADC) analysis using MS | ADC DAR analysis/ Drug to Antibody ratio and intact mass | |
Intact mAb & fragment analysis | Light chain (LC) and heavy chain (HC) analysis Fragment (Fab and Fc) analysis scFc and F(ab’)2 analysis |
Learn how to easily produce linear pH gradients for fast and robust therapeutic protein separations pH gradient buffer kits with ion exchange columns.
Learn how to optimize the challenging characterization of proteins, mAbs, biosimilars, and their charge variants using new ProPac Elite weak cation exchange column chemistry.
Collaboration between NIBRT and Thermo Fisher Scientific is designed to enhance structural knowledge, accelerate innovations, and ensure drug safety and quality.
Unique applications compendium from NIBRT and Thermo Fisher Scientific focused on simplified, faster methods that deliver highly informative characterization data.
Learn more about the innovative technologies for charge variant analysis.
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