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Speciation Analysis of Plasma Proteins using Asymmetric Flow FFF-ICP-MS

Postnova Analytics GmbH

New application study demonstrates the plasma protein analyzing abilities of Postnova Analytics’ AF4-ICP-MS system

12 Jun 2016
Weylan Kiam-Laine
Microbiologist

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Postnova Analytics reports on exciting advances in the analysis of plasma proteins using a hyphenated system that couples asymmetric flow Field-Flow Fractionation (AF4) and inductively coupled plasma-mass spectrometry (ICP-MS). In a new application study researchers show how an AF4-ICP-MS system is able to obtain valuable information on the multi-element speciation and absolute molecular mass of human plasma proteins such as Albumin and Transferrin.

Human Serum Albumin (HSA) and Transferrin are the two main plasma proteins that are known to bind and transfer metal ions in biological fluids. Metal-protein complexes are involved in many health studies such as metal-based drugs for cancer or diabetes treatments. Asymmetric flow field-flow fractionation (AF4) is very similar to liquid chromatography, and has been successfully applied to protein fractionation in several biological applications. However unlike liquid chromatography for which fractionation occurs by interaction with a stationery phase, AF4’s lack of stationery phase offers improved recovery for a range of species from complex biological matrices. Further the AF4 technique has been able to achieve this (in some cases) using non-denaturing carriers, which offers significant benefits for the separation of unstable protein metal complexes.

In the application study, a Postnova AF2000 Multiflow Mid Temperature Field-Flow Fractionation system coupled with an Agilent 7700 ICP-MS system was used to separate a mixture of Human Serum Albumin binding 63Cu and Transferrin binding 56Fe metal ions. The AF2000 system separated the proteins by size and the ICP-MS system detected the metal ions to obtain metal distributions of the protein mixture. The researchers demonstrate that AF4-ICP-MS is an excellent tool for the non-denaturing and element-selective detection of plasma proteins.

The demonstrated benefits of the AF4-ICP-MS technique show particular promise for demanding applications in which the two plasma proteins under study act as binders of the same element (e.g. vanadium in Diabetes studies). Other applications under investigation by Postnova, using the AF4-ICP-MS technique, include patient physiology studies involving identification of metal uptake in plasma ; investigation and optimization of therapeutic protein and antibody production ; detection of protein contamination and quality control of therapeutic protein and antibodies.

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Mass SpectrometryMass spectrometry (MS) is a powerful analytical technique used to identify and quantify molecules based on the mass-to-charge ratio of gas-phase ions. It provides detailed information about the structure, composition, and properties of compounds and is widely used across fields such as environmental monitoring, materials science, drug discovery and development, food and beverage testing, and wider chemical research. Key MS techniques include tandem mass spectrometry (MS/MS), liquid chromatography–mass spectrometry (LS-MS) and inductively coupled plasma (ICP-MS). Choosing from these wide range of techniques and technologies can be a daunting task, so keep up to date with scientific applications, performance expectations, and customer reviews here all in one place. Visit our product directory to receive quotes direct from the manufacturer. AntibodiesAntibodies are used in techniques such as confocal and fluorescence microscopy, flow cytometry, ELISA, ELISPOT, immunohistochemistry, western blotting and immunopreciptation. Select specific antigen reactivity, high specific affinity, low non-specific binding, monoclonal or polyclonal, primary or secondary antibodies and associated conjugates such as an enzyme or dye for visualization.ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Bioprocessing / FermentationBioprocessing is the use of biological materials to perform commercial, scientific or medical research processes. Biological materials used include cells, enzymes and organisms. Usually bioprocessing requires a batch or continuous bioreactor such as a fermentor or cell culture system. The advantages of using a reactor include high productivity, easy configuration, adjustable values and automation.Protein BiologyThe analysis of protein expression, identity and function is vital for many areas of life science research and drug discovery. Some of the most commonly used techniques in protein analysis include Western blotting, electrophoresis and mass spectrometry.Molecular Weight AnalysisMolecular weight analysis determines the size and structure of biomolecules, essential in protein characterization, polymer studies, and drug development. Techniques like mass spectrometry ensure precise results. Compare solutions for molecular weight analysis in our peer-reviewed product directory.Human Serum AlbuminICP-MSInductively Coupled Plasma Mass Spectrometry (ICP-MS) is a powerful analytical technique used to determine the concentrations of elements and their isotopes in a variety of samples, including water, soil, and biological tissues. It involves ionizing elements in the samples with extremely high-temperature Argon (Ar) plasma and then using a mass spectrometer to measure the number of ions based on their mass-to-charge ratio (m/z). Explore ICP-MS systems in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Elemental AnalysisElemental analysis involves determining the elemental composition of a sample, often used in environmental, pharmaceutical, and material sciences. Techniques like ICP-MS, X-ray fluorescence, and atomic absorption spectroscopy allow precise quantification of elements such as metals and nonmetals in complex matrices. Browse our peer-reviewed product directory to find the best elemental analysis tools, compare products, check reviews, and get pricing directly from manufacturers.SpeciationPlasmaProcess OptimizationProcess optimization involves improving the efficiency, cost-effectiveness, and quality of processes in various industries, including manufacturing, pharmaceuticals, and biotechnology. Explore process optimization solutions in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Antibody ProductionAntibody production is essential for a wide range of applications in biochemistry and immunology. Ensuring high-quality monoclonal and polyclonal antibodies are produced is key. Explore antibody production tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.

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Speciation Analysis of Plasma Proteins using Asymmetric Flow FFF-ICP-MS