Quick Albumin Removal Kit

Introduction

Human serum or plasma contains a wealth of proteomic information and is frequently analyzed for protein biomarkers of normal and pathological states. The extensive concentration range of proteins contained in human serum is one of the most challenging obstacles in studying it. Proteins that are abundantly expressed, such as albumin and IgG, account for 70 percent of total protein in serum/plasma. In contrast, protein biomarkers may present considerably lower amounts (pg/ml). High abundance proteins pose problems for analytical procedures like one-dimensional and two-dimensional electrophoresis, high-performance liquid chromatography, and mass spectroscopy because they hide the less abundant proteins of interest. High abundant proteins must be effectively, reproducibly, and extracted explicitly from blood samples to correctly detect the lower abundant proteins. The selective removal of albumin from human samples (e.g., plasma, serum, urine, or cerebrospinal fluid) is necessary before performing proteome analysis, particularly when using mass spectrometry.

Although Cibacron Blue F3GA, an affinity dye ligand, was covalently bonded onto commercially available supports for HSA adsorption from both aqueous solutions and human plasma, it lacks selectivity. In immunoaffinity systems, anti-albumin antibodies are also employed. These techniques have reasonable specificity in general, but they are costly. Furthermore, because many of these systems are based on agarose resin or its derived column, their procedure for the enrichment of the less abundant proteins is time-consuming, unable to handle large samples volume in a short time and challenging to adapt to the automation system. At the same time, they are frequently performed by increasing the sample volume, requiring an additional protein concentration step. New albumin removal methods with improved efficiency, homogeneity, and throughput are required to address the above issues.

Hygia Quick Albumin Removal Kit is based on our unique magnetic beads modified with our proprietary chemistry for quick and easy albumin depletion from serum and plasma samples. The beads bind to all other proteins in the sample (except albumin) in particular buffer conditions, allowing the binding and release of all less abundant proteins but albumin. The enriched and eluted proteins can be used for downstream applications such as proteome analysis, biomarker detection, enzyme assays, SELDI analysis, protein array pixelation, 1D and 2D gel electrophoresis, LC/MS, and MALDI-TOF MS.

Workflow

Work with the Albumin Removal Kit is straightforward. Mix the beads with serum or plasma samples and incubate them with continuous rotation. The beads remain suspended in the sample solution during mixing, allowing all other proteins in the sample (except albumin) to bind to the beads. After incubation, the beads are collected and separated from the sample using a magnet rack. Then the bound proteins are eluted.

Features and Advantages

● Quick, Easy, eliminates columns, centrifuge, or filters.

● Remove >95% Albumin

● High throughput: Rapidly process up to 96 samples in less than 30 minutes

● Low abundance enrichment equal to or better than hexapeptides or antibodies

● Mild elution preserves the tertiary structure and allows for easy transfer to secondary analysis

● Scalable – easily adjusts for sample size and automation

● Minimal sample dilution

● Suitable for LC-MS, activity-based protein profiling, and proteomic research