ProteomeLab™ IgY-12 High Capacity Primate-Optimized Proteome Partitioning Kits

Albumin IgG Transferrin Fibrinogen IgA α2-macroglobulin IgM α1-Antitrypsin Haptoglobin Orosomucoid Apolipoprotein A-I Apolipoprotein A-II The ProteomeLab IgY-12 Proteome Partitioning kits target these twelve highly abundant proteins.

The ProteomeLab IgY-12 High Capacity Proteome Partitioning kits are specifically designed to remove twelve highly abundant proteins from human/primate biological fluids such as serum and plasma.

This provides an enriched pool of lower abundant proteins, free from interference by the removed proteins, ready for discovery and validation of biomarkers, drug targets, and therapeutic proteins and antibodies.

Kits are available in both spin column and LC column formats, depending upon the desired capacity per cycle. The observed capacity of human or other primate serum or plasma varies from 20 µL to 250 µL depending on the kit selected (see back). Our IgY (avian) based technology uses physiological buffers for binding and washing, and avoids urea and other chaotropic agents for elution that can precipitate at low temperature.

Our ProteomeLab IgY-12 spin column kit is specifically designed to work in combination with the ProteomeLab SP Sample Preparation system, while the higher capacity LC kits are designed to work with the ProteomeLab PPS Sample Preparation system.

The lower abundant proteins in the flow-through fraction and the highly abundant proteins in the bound/eluted fractions can be further fractionated with the ProteomeLab PF 2D system, advancing the detection and identification of low abundant proteins.

ProteomeLab IgY High Capacity primate-optimized 12-plex proteome partitioning kits.

The ProteomeLab IgY-12 High Capacity SC, IgY-12 LC2, and IgY-12 LC10 Proteome Partitioning kits are selected based on the following:

• the volume of human / primate biological fluids needed to yield the target quantity of partitioned protein for subsequent analysis
• the sample throughput requirements

IgY-12 High Capacity SC Spin Column kit The IgY-12 SC Spin Column kit is based on centrifugation and has a capacity of 20 µL human or primate serum or plasma per spin column cycle. The expected yield of a sample partitioned of the 12 highly abundant proteins is about 160 µg per cycle. We recommend the use of the ProteomeLab SP Sample Preparation System with the IgY-12 SC kit.

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IgY-12 High Capacity LC2 Column Kit The IgY-12 LC2 column kit requires liquid chromatography equipment and has a capacity of 50 µL human or primate serum or plasma per LC column cycle. The expected yield of a sample partitioned of the 12 highly abundant proteins is about 400 µg per cycle. We recommend the use of the ProteomeLab PPS Proteome Partitioning System with the IgY-12 LC2 column kit.

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IgY-12 High Capacity LC10 column kit The IgY-12 LC10 column kit also requires liquid chromatography equipment and has a capacity of 250 µL human or primate serum or plasma per LC column cycle. The expected yield of a sample partitioned of the 12 highly abundant proteins is about 2 mg per cycle. Samples can be pooled for ProteomeLab PF 2D (recommended 1 - 5 mg) or other fractionation methods and can be analyzed by 2D electrophoresis or LC-MS methods. We recommend the use of the ProteomeLab PPS Proteome Partitioning System with the IgY-12 LC10 column kit.

The above formats have been integrated into the ProteomeLab IgY spin column and ProteomeLab high capacity solutions.

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Cleaner Capture and Broader Antigen-Binging Host Range

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Figure 1: Since the polyclonal IgYs are avian, the Fc region of the antibody does not bind mammalian complement factors, Rheumatoid Factor, IgM, Fc receptor and Protein A or G, a known issue related to capture with mammalian antibodies. Coupled with high specificity across multiple epitopes, you get very specific binding across species while achieving very clean capture with little background. Firgure 1 highlights whole serum run by SDS-PAGE in lane #1, serum with just albumin partitioned out by Proteome-Lab IgY-HSA SC chemistry in lane #2, and the isolated/partitioned albumin captured cleanly in lane #3. The partitioning efficiency of targeted highly abundant proteins from human plasma is highlighted in Table 1 below. Additionally, due to the phylogenetic difference, mammalian proteins are often more immunogenic in birds than in mammals, hence the broader antigen-binding host range. ProteomeLab IgY Chemistry effectively leverages this well-documented evolutionary advantage.

Figure 2: 2DE protein fingerprinting of human serum before and after ProteomeLab IgY-12 partitioning. (A) Unfractionated human serum proteins before ProteomeLab IgY-12 treatment, (B) Human serum protein fraction after ProteomeLab IgY-12 treatment partitioned off the 12 most abundant proteins. (C) Proteins eluted from ProteomeLab IgY-12 column. 100 µg of protein was loaded on each gel.

Figure 3: ProteomeLab PF 2D pI/hydrophobicity maps of human serum before and after ProteomeLab IgY-12 partitioning. (A) Unfractionated human serum proteins before ProteomeLab IgY-12 treatment, (B) Human serum protein fraction after ProteomeLab IgY-12 treatment partitioned off the 12 most abundant proteins. (C) Proteins eluted from ProteomeLab IgY-12 column. 2.5 mg of protein was loaded in each PF 2D run. The ProteomeLab PF 2D fractionates a proteome into hundreds of liquid phase fractions and generates differential displays for the comparison of these proteomes. By coupling the enrichment power of the IgY-12 proteome partitioning with the fractionation power of the ProteomeLab PF 2D you will vastly enhance the discovery and validation of biomarkers and drug targets from biofluids such as serum and plasma.

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ProteomeLab IgY-12 Kits