Waters - Protein Reversed-Phase Columns
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Reversed-Phase Chromatography
Reversed-phase chromatography of proteins, done with columns consisting of wide-pore size particles ( e.g, 300Å) functionalized with short-ligand length chemistries, is a separation technique based on the relative hydrophobic characteristics of the proteins in solution. Gradients of increasing organic solvent concentration are frequently used to affect separations in the presence of ion-pairing reagents(e.g., 0.1% TFA) that minimize undesired ionic interactions. In general, the hydrophobicity of the protein or protein sub-unit determines the elution order, with the least hydrophobic proteins eluting first. Factors such as particle composition (silica vs hybrids), pore size, ligand type and density, as well as separation conditions (e.g., gradient duration, separation temperature, flow rate) all play an important role in obtaining a separation that meets application requirements.
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Features of Protein Reversed-Phase Columns
[u]BEH Particle Technology[/u]
Designed for Protein Separations
The separation of large molecular weight biological compounds can be challenging. For more than 20 years, chromatographers have benefited from performance characteristics that Waters reversed-phase columns have provided for protein separations. Waters Ethyl Bridged Hybrid (BEH) Particle Technology can help overcome performance limitations experienced using 100% silica-based materials for challenging protein separations.
300Å pores
C4 ligand
Minimal undesirable secondary interactions
Minimal detectable carryover at elevated temperature separations
[u]Reproducible Protein Separations[/u]
Well characterized, state-of-the-art bonding procedures
Low pH and high temperature stability
Quality-control tested with diverse protein mixture
Consistent batch-to-batch reproducibility
Available 1.7 Μm particles for UPLC and nano UPLC applications
zoom batch to batch reproducibility
[u]Protein Separation with nano- and microflow LC-MS[/u]
Microflow LC-MS for protein analysis is greatly simplified with the 150 µm iKey Separation device due to plug-and-play design and fitting-less fluid connections. The iKey offers an up to 40x increase in sensitivity compared to 2.1mm ID UPLC.
For nanoflow LC-MS protein analysis, ACQUITY UPLC M-Class Columns are specifically designed for low dispersion nano LC. The nano- to microscale LC/MS ACQUITY UPLC M-Class Columns and trap columns enable nano- and microscale separations under UPLC conditions at 15,000 psi, fully leveraging the separation power of sub-2-µm particle technology.
[u]BioSuite pPhenyl Reversed-Phase Chromatography (RPC) HPLC Columns[/u]
BioSuite RPC Column offerings include a phenyl (pPhenyl) chemistry bonded to a pH stable, methacrylic ester-based polymeric resin. The 1000A pore size of the pPhenyl base matrix accommodates proteins up to 5,000,000 Daltons.
The pPhenyl RPC chemistries are available in 21.5 x 150 mm columns for “lab-scale” isolations while a 2.0 x 75 mm column is well suited for narrow-bore HPLC and LC-MS applications.
[u]Symmetry300 C4 HPLC and UHPLC Columns[/u]
Symmetry300 C4 particles are 100% silica-based and are synthesized using ultrapure organic reagents resulting in high-purity material with very low silanol activity for outstanding peptide and protein separations and recoveries.
300A pore for peptide and protein applications
Fully end-capped to minimize undesired secondary interactions
Alternative separation selectivity compared to Waters BEH C4, 300A hybrid material
QC tested with peptide samples to help ensure excellent batch-to-batch consistency
[u]Delta-Pak Columns[/u]
Delta-Pak HPLC Columns are based on highly stable, bonded, endcapped 5 Μm or 15 Μm spherical 100% silica-based particles. These columns are offered in two different pore sizes -- 100Å or 300Å -- with either C4 or C18 ligands to address different selectivity and retentivity needs. They are available in different 15 Μm cartridge and column configurations, providing consistent and predictable scale-up capabilities for milligram-to-gram scale purification.
[u]MassPrep Protein Standard[/u]
This intact protein validation mix is designed for verification of HPLC/UPLC instruments. This standard is also used to evaluate the performance of columns used for protein analysis, specifically the ACQUITY UPLC and XBridge Protein BEH C4 Columns. The MassPrep Protein Standard contains proteins that have a wide range of isoelectric points, molecular weights, and hydrophobicities.
General Specifications
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