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HPLC Analysis

At CellMosaic, we have tremendous experience in analyzing and purifying a variety of small organic compounds, biopolymers, and bioconjugates by liquid chromatography utilizing different media and columns. The equipment we use is an Agilent 1100 HPLC system with automatic sample injector, fraction collector, and diode array detector. We have a variety of columns to use and established standard HPLC methods for analyzing peptides, oligos, proteins, antibodies, and some bioconjugates. For the development of HPLC methods involving special molecules, we encourage you to talk with our specialist.

  • Reverse-phase HPLC analysis: This type of analysis involves HPLC/UV analysis of small organic compounds, peptides, oligos, antibodies, enzymes, proteins, and their conjugates. We help customers develop HPLC methods for analyzing bioconjugates, such as proteins, peptides, oligos, and antibody-drug conjugates, among others. We have a variety of reversed phase columns, including C4, C8, C18, and phenyl-modified columns, with which to perform method development. Generally, we can narrow down the type of column and buffers needed for separation based on the nature of the analyte.
  • Size-exclusion chromatography: We have a variety of gel filtration columns from Tosoh Bioscience, Phenomenex, Applied Biosystems, and GE life science. This chromatographic method is suitable for analyzing mixtures of biopolymers, separating the components according to size. It can be used to determine the approximate molecular weight of the molecules. In the case of bioconjugate analysis, it can be used to assess the amount of unlabeled molecules and degree of bioconjugate aggregation in a sample.
  • Hydrophobic interaction chromatography (HIC): This chromatographic method is good for analyzing proteins or antibody/protein bioconjugates based on their hydrophobicity. The theory behind hydrophobic interaction chromatography is similar to reverse-phase chromatography; however, the degree of substitution of HIC adsorbents is diluted compared to reverse-phase chromatography. Thus, the compound can be eluted in a more polar and less denaturing environment. For example, HIC has been used extensively to analyze drug loading in antibody-drug or protein-drug conjugates. At CellMosaic, we have a selection of HIC columns from YMC to perform such analyses.
  • Hydrophilic interaction chromatogrpaphy (HILIC): This chromatographic method is good for analyzing small molecules that are very hydrophilic and will not retain on reversed phase columns, such as amino acids and hydrophilic metabolites containing polar charged groups. On the other hand, if a compound is so hydrophobic that it is insoluble in aqueous buffer, one can use >95% organic buffer and a HILIC column to conveniently analyze the molecules. At CellMosaic, we have a selection of HILIC columns to perform such analyses.
  • Anion and cation exchange chromatography: This type of chromatography uses the charged state of the molecule to achieve separation. It is generally a mild and selective method for analyzing peptides, oligos, or proteins based on the charge. We offer such analyses at a customer’s request.

Pricing Information for Some Standard HPLC Services

CodeService TypeDescriptionSetup Fee/Rate
Purity assay (Std. HPLC)
Standard C18 reverse phase chromatography for oligos, peptides, and small size proteins.
Purity plus UV characterization (Std HPLC)
Standard C18 reverse phase chromatography plus the UV spectra of individual peaks.
Purity assay (size exclusion)
Size exclusion column chromatography for large biomolecules and bioconjugates
Purity plus UV (size exclusion)
Size exclusion column chromatography plus the UV spectra of individual peaks.
Molecular weight determination (Size exclusion)
This analysis uses size exclusion columns and compares the HPLC data with the standard MW curve to determine the approximate molecular weight.
Hydrophobic interaction HPLC (HIC)
This analysis uses hydrophobic interaction chromatography to analyze protein-drug and antibody-drug conjugates.
Anion or cation HPLC
Non-standard HPLC that uses an anion or cation stationary phase.
Hydrophilic interaction HPLC (HILIC)
This analysis uses hydrophilic interaction chromatography to analyze small organic compounds.

HPLC: high performance liquid chromatography.

Note: Price is per Injection. Standard service is 1-3 days. Please ask us for a quote before placing an order.