Ion-Exchange Chromatography

PolyCAT A™ is a high-performance weak cation-exchange (WCX) material that delivers exceptional results for protein and peptide analysis. Its unique manufacturing process ensures sharp peaks, minimal tailing, and high binding capacity, resulting in efficient separations and recoveries.

Key Advantages:

• Excellent resolution for complex protein and peptide mixtures
  
   
• Versatile applications, including protein variant analysis, hemoglobin separation, monoclonal antibody analysis, and histone modification profiling 
  
   
• Compatible with a wide range of mobile phases, including volatile solvents for mass spectrometry analysis
  
  

Protein Variant Analysis Example: HGH

PolyCAT A™ can resolve many intact proteins differing by a single residue. The following figure shows the difference in the degradation pattern of recombinant human growth hormone (HGH) incubated at two different pHs. The desPhe-Pro variant, a nonenzymatic cleavage product, is well separated from HGH, as are its corresponding variants. Such studies are critical in determining which pH yields the most stable formulation.

Histone Modification Analysis

PolyCAT A™ has proven valuable in histone modification analysis, enabling researchers to explore the complex interplay of modifications on gene expression and chromatin structure. This has led to significant advancements in our understanding of epigenetic regulation and its impact on various cellular processes.

PEGylation Analysis

PolyCAT A™ columns excel in separating positional variants of PEGylated proteins, which is crucial for assessing their biological activity and therapeutic efficacy. The chromatogram below showcases the separation of PEGylated variants of a neurotrophic factor, where the main product and minor variants with differing PEGylation sites are clearly resolved.  

PolySULFOETHYL A™ is a strong cation-exchange (SCX) material specifically designed for HPLC of peptides. Its unique hydrophilic properties minimize hydrophobic interactions, leading to high recovery and reduced peak tailing.

Key Advantages:

• Exceptional separation capabilities for complex peptide mixtures, including tryptic digests in proteomics analyses
• Selective isolation of specific peptide classes, such as phosphopeptides, disulfide-linked peptides, and C-terminal fragments
• High capacity for improved retention and fractionation of weakly basic peptides

SCX-RPC of Tryptic Digests

The combination of SCX and Reversed-Phase Chromatography (RPC) offers a powerful approach for multidimensional HPLC of peptide mixtures. This technique allows for the separation of complex tryptic digests into fractions by differences in charge-to-mass ratio, enabling the identification of a vast number of peptides in a sample.

Isolation of Phosphopeptides and Other Classes of Peptides from Tryptic Digests

PolySULFOETHYL A™ columns excel in the selective isolation of specific peptide classes from tryptic digests. The 200-Å pore version of PolySULFOETHYL A™ is particularly effective in separating phosphopeptides from non-phosphorylated peptides due to its higher surface area.

PolyWAX LP™ is a weak anion-exchange (WAX) material that offers exceptional versatility for the separation of a wide range of biomolecules. Its unique properties make it ideal for applications where traditional reversed-phase chromatography falls short.

Key Advantages:

• High-resolution separations of polar and charged molecules, including proteins, peptides, and oligonucleotides
• Exceptional selectivity for the isolation of acidic peptides and post-translationally modified peptides
• Compatible with a variety of mobile phases, including those used in hydrophilic interaction chromatography (HILIC)

Protein Phosphorylation Analysis

PolyWAX LP™ exhibits exceptional selectivity in separating proteins that differ by a single phosphate group. This capability is crucial for studying protein phosphorylation, a ubiquitous post-translational modification that regulates various cellular processes.

Oligonucleotide and DNA Analysis

PolyWAX LP™ is the preferred method for high-resolution separation of oligonucleotides and their analogs, especially those with more than 15 bases. It also offers a faster and more convenient alternative to PAGE gels for purifying double-stranded DNA products from PCR reactions.

Additional Applications

PolyWAX LP™ is also highly effective in the following applications:

• Purification of acidic proteins and polypeptides from natural products
   
• Predigest fractionation of intact proteins via mixed-bed ion-exchange in proteomics   
   
• Anion-exchange of small, acidic solutes  
   

PolySAX LP™ is a silica-based strong anion-exchange (SAX) material with an adsorbed, cross-linked coating. It offers high capacity for both small and large solutes, making it a powerful tool for challenging separations.

Key Advantages:

• High resolution for the separation of complex mixtures of proteins, peptides, and other charged biomolecules
• Exceptional selectivity for the isolation and purification of negatively charged compounds
• Can be used in AEX and ERLIC-SAX modes

Featured Publication: Unveiling the Complexity of Non-Canonical Phosphorylation

A recent study published in The EMBO Journal utilized PolySAX LP™ to enrich and fractionate phosphopeptides, enabling the identification of a diverse range of non-canonical phosphorylation sites on human proteins. This research highlights the versatility of PolySAX LP™ in advancing our understanding of protein phosphorylation and its role in cellular regulation.