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Using Cleanascite™ as a Lipid Absorption and Clarification Reagent


Validation of a proteomic sample preparation technology, Cleanascite™, as a simple, yet effective high binding capacity for lipids with minimal cross-reactivity with proteins.


Background

BSG supplies an environmentally sound replacement for chlorinated/fluorinated hydrocarbons (eg. freon) to help purify and analyze antibodies, recombinant proteins, nucleic acids, and proteoglycans. Cleanascite™ selectively removes lipids, cell debris, lipoproteins, as well as impurities from lipemic serum/plasma, Cohn paste, transgenic milk, egg yolk and biological samples. It is ideal for pretreatment of ascites, serum, cell & tissue culture, bile, and organ homogenates, prior to macromolecule analysis or further purification.

Cleanascite™ has been extensively cited in over 30 journal articles. In this case report, we describe its use in:

Steven W. Taylor, Nigel J. Clarke, Zhaohui Chen, Michael J. McPhaul. A high-throughput mass spectrometry assay to simultaneously measure intact insulin and C-peptide. Clinica Chimica Acta. January 2016, doi:10.1016/j.cca.2016.01.019


The Challenge

When removing lipid interferences, it’s essential to do so without compromising the quality or quantity of biomarkers to be measured. Quality assurance in the purification and analysis process is a key consideration.

In this particular study, simultaneously measuring intact insulin and proinsulin derived C-peptide, was needed to help predict development of diabetes mellitus, as well as in differential diagnosis in cases of hypoglycemia. There was necessity for removing lipids without compromising the quantity or quality of these 2 biomarkers. In this study, Cleanascite™ is shown both to improve LC-MS measurements of these biomarkers, and validated in accordance with CLIA ’88 guidelines.


The Solution

As one of the main advantages of Cleanascite™, it offers a simple and efficient proteomic sample preparation technology for clearing lipid-associated matrix effects from human sera.

The article states “…15 µl of internal standard were added to each well followed by 50 µl of Cleanascite™ delipidation reagent previously mixed into a uniform suspension by a brief aspiration/dispense cycle within its reagent reservoir.” The article further notes a key component of the methodology as “…the use of a delipidation reagent to enhance immunocapture…The result was greatly enhanced recoveries and tighter CVs for the IS {internal standard} throughout the plate”.

Additional key advantages of Cleanascite™ include:

  • Extends the life of membrane and chromatographic columns.
  • Ideal for delipidation treatments for downstream processing of large-scale therapeutic proteins, enzymes and antibodies.
  • The reagent is supplied as a ready to use, solid-phase suspension in saline.

The Outcome

Without a reliable lipid removal reagent and clarification like Cleanascite™, the study could have been compromised due to matrix effects or cross-reactivity.


BSG Featured Advantage: Consumable

  • Cost-effective, not derived from biologicals
  • No specialized instruments or HPLC
  • Consumable easy to use suspension format
  • No regeneration, so no prep to prep variability.

This article along with these notable 2017 citations, illustrate the importance of having an effective and reliable sample prep method for removal of lipids, in the purification and analysis of antibodies, bile proteins, nucleic acids, proteoglycans, and other macromolecules.


Notable 2017 References

View all of our references