ResourceSeparations

Protein A Affinity IgG Capture Followed by AdvanceBio SEC Aggregation Analysis

25 Sept 2017

This application note describes the quantification of aggregates in biotherapeutic samples using the Agilent InfinityLab 2D-LC solutions. Protein A affinity capture was performed in the first dimension, and size exclusion chromatography in the second dimension. Multiple heart-cutting was used to transfer the samples from the first to the second dimension. This technique enabled the entire characterization to be done with a single analysis, eliminating the need for fraction collection and multiple injections.

Agilent 1290 Infinity III 2D-LC System

Agilent Technologies

Increase resolution where you need it with the 1290 Infinity III 2D-LC System, which offers comprehensive two-dimensional liquid chromatography (2D-LC), (multiple) heart-cutting 2D-LC analysis, and high-resolution sampling. This system allows you to use 2D-HPLC for making buffer separations compatible to MS detection, while easy-to-use software enables fast method setup and refined data analysis.

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UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Size Exclusion ChromatographySize exclusion chromatography (SEC) is a liquid chromatographic technique used to separate and identify molecules based on their size, including polymers, proteins, nanoparticles & nucleic acid. SEC is further subdivided into gel filtration chromatography (GFC) and gel permeation chromatography (GPC) when an aqueous or organic mobile phase is used respectively. HPLC-based SEC systems and columns are available for analytical separation alongside SEC spin columns or filters to prep samples for many protocols. Find the best size exclusion chromatography equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.LC-MSLC-MS (liquid chromatography-mass spectrometry) systems and equipment are used for separation and quantitative analysis of complex mixtures, combining liquid chromatography and mass spectrometry. Quantify proteins, contaminants, pesticides or screen for drug metabolites with a high level of sensitivity. LC-MS systems and equipment include reverse phase, normal phase and specialized columns integrated with various MS detectors such as time-of-flight (TOF), quadrupole, orbitrap or ion trap mass analyzers. LC-MS/MS instruments equipped with a qTOF or triple quadrupole analyzer give greater sensitivity and resolving power to your analysis. Find the best LC-MS equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Protein QuantificationThe detection and quantification of proteins in a sample is vital across life sciences, pharmaceutical research and clinical diagnostics, and a variety of equipment is available to scientists to simplify the workflow. Proteins of interest can be easily labeled and detected on light-based detection instruments. Immunoassay kits allow you to identify a specific protein of interest and protein detection beads or antibody microarrays allow you to identify multiple specific proteins at once. Protein interactions and enzyme activity can also be monitored with protein-protein interaction assays. Additionally, the biophysical characterization of proteins is made easy with biokinetic analyzers. Find the best protein detection and quantification products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.BiotherapeuticsBiotherapeutics are proteins and other compounds (such as nucleic acids) produced by living organisms that have uses as therapeutics or in <i>in vivo</i> diagnostics. The most well-known example of a biotherapeutic product, and the first to be approved for therapeutic use, was recombinant human insulin.2D ChromatographySECSize exclusion chromatography (SEC) separates molecules based on size, ideal for protein purification, polymer analysis, and biopharma applications. SEC ensures precise characterization of biomolecules for research and industry. Explore peer-reviewed SEC products and compare options for your lab.LC-MSLiquid Chromatography-Mass Spectrometry (LCMS) is a powerful analytical technique that combines the separation power of liquid chromatography with the detection capabilities of mass spectrometry. It is widely used for qualitative and quantitative analysis of complex mixtures in pharmaceuticals, proteomics, and environmental studies. Browse our peer-reviewed product directory to find the best LCMS systems, compare products, check reviews, and get pricing directly from manufacturers.
Protein A Affinity IgG Capture Followed by AdvanceBio SEC Aggregation Analysis