 |
 |
 |
|
Albumin and IgG Depletion From Serum/Plasma for Proteomics
- IgG removal >90% (70-80% of total Immunoglobulins removed)
- Albumin removal >95%
- Seamless and simple < 1 hour protocol
- Low abundance enrichment equivalent to immuno-affinity
- Disposable, cost-effective, no column regeneration or cross-contamination
- Works for most species tested including human, sheep, rat, mouse, bovine
- On-bead protocols improve workflow and efficiency, especially suited to targeted proteomics
- Suitable for LC-MS, 1 and 2D Gels, ELISAs, Enzyme and other Functional Assays.
The classical plasma proteins generally fall into functional categories, forming the vast majority of the mid-to-high abundance proteome. In serum, these sub-proteomes by mass content are: Albumin 50-60%; Immunoglobulins 10-20%; Transport Proteins (Transferrin, Apo) 5-10%; Complement related Proteins 3-5%; Protease Inhibitors 2-5%; and all others 2-5%. While these sub-proteomes are required for normal body homeostasis, they nevertheless can become dysfunctional during acute-phase and chronic stimuli.
So, depending on the needs of the investigation, it can be valuable to consider that one or more of these categorical sub-proteomes is simply background noise whereby depletion is beneficial. While in other cases, these same categorical sub-proteomes might provide new data and information and consequently, should not be depleted. Different AlbuVoid™, AlbuVoid™ PLUS and AlbuSorb™ PLUS workflows support different proteomic biases as outlined in the following Table.
Click Here For The AlbuVoid™ Plus Product Sheet.
Products and digest conditions produce different sub-proteome windows of observation. So, depending on the needs of the investigation, it can be valuable to consider that one or more of these categorical sub-proteomes is simply background noise whereby depletion is beneficial. While in other cases, these same categorical sub-proteomes might provide new data and information and consequently, should not be depleted. Categorically the acute-phase sub-proteomes differentiated in disease may vary greatly from those associated with chronic sub-proteomes. So there is great benefit in having options to enrich or deplete one or more of these sub-proteomes.
BSG's Albumin and IgG Removal Kits offer many such options:
- The 'PLUS' products substantially deplete Immunoglobulins through separations at the protein level.
- The variable regions of Immunoglobulins are extremely heterogeneous, generating a background noise across the full LC gradient. On-bead digestion (BASP™) with AlbuVoid™ substantially reduces the influence of such Ig peptide features. So in addition to workflow simplicity, BASP™ can be advantageous utilized in targeted proteomic workflows whenever the target proteins do not require strong denaturing conditions.
- With the exception of Immunoglobulins whereby FASP generates many more spectral features, both strong denaturing conditions (FASP) and on-bead digest (BASP™) conditions produce similar protein profiles.
- Both Apolipoproteins and heavily glycosylated proteins (i.e., α1-Acid Glycoprotein) bind poorly to AlbuVoid™. For quantitative studies within these classes of proteins, AlbuSorb™ PLUS is recommended.
- The Complement sub-proteome is especially enriched by AlbuVoid™ PLUS. The digest conditions may bias towards one or more functional sub-populations, likely due to conformational transitions and protein-protein interactions (i.e., Factor Bb, Properdin) that occur upon activation. This needs further investigation.
- The low abundance sub-proteome is enriched 5+ fold with AlbuVoid™ and 4+ fold with AlbuSorb™ PLUS.
|
Product
|
Size
|
# Serum Preps
|
Item No.
|
|
AlbuVoid™ PLUS Kit
|
10 preps
|
10, 25µl Serum samples
|
NP-AVK-10
|
|
Items
|
Item No.
NP-AVK-05
|
Item No.
NP-AVK-10
|
Reagents
|
|
Kit#1 - IgG Depletion Kit
|
|
|
|
|
NuGel™ Protein A Beads (NP)
|
300mg
|
600mg
|
Supplied
|
|
Buffer 1
|
5 ml
|
10 ml
|
Supplied
|
|
Spin-X Centrifuge tube Filters
|
5
|
10
|
Supplied
|
|
Items
|
Item No.
NP-AVK-05
|
Item No.
NP-AVK-10
|
Reagents
|
|
Kit#2 - Albumin Depletion Kit
|
|
|
|
|
AlbuVoidTM Beads (AVK)
|
125mg
|
250mg
|
Supplied
|
|
Binding Buffer AVBB pH 6.0
|
3 ml
|
6 ml
|
Supplied
|
|
Wash Buffer AVWB pH 7.0
|
12 ml
|
24 ml
|
Supplied
|
|
Elution Buffer AVEB pH 10
|
2 ml
|
4 ml
|
Supplied
|
|
Spin-X Centrifuge tube Filters
|
5
|
10
|
Supplied
|
|
DTT, Iodoacetamide, Trypsin and Formic Acid
|
|
|
Not Supplied
|
|
|
|
 |
|
 |
|
|
|
 |
|
 |
|
References
-
AlbuVoid™ PLUS & AlbuSorb™ PLUS -
Evaluating Different Windows of Observation Solves The Many Challenges of Serum Proteomics
Zheng H, Soherwardy A, Roy S, Kuruc M, Avadhani S. AlbuVoid™ Enrichment & Antibody Depletion – Tackling the Challenges of Serum Proteomics Part II. Poster reprint first presented at 26th International Molecular Med TRI-CON, March 10-15, 2019, San Francisco, CA USA
Vialaret, Jerome & Kadi, Sarah & Tiers, Laurent & O Flynn, Robin & Lehmann, Sylvain & Hirtz, Christophe. (2018).
Albumin depletion of human serum to improve quantitative clinical proteomics. Current Topics in Peptide & Protein Research 19. 53-62.
Poillet-Perez, Laura, et al. "
Autophagy maintains tumour growth through circulating arginine." Nature (2018): 1.
Roy, Swapan, and Matthew Kuruc. "
Methods to Monitor the Functional Subproteomes of SERPIN Protease Inhibitors." Functional Proteomics. Humana Press, New York, NY, 2019. 41-54.
Mihara, Keisuke, et al. "
Identification of Specific Protein Markers of Rheumatoid Arthritis in Synovial Fluid and Serum." Journal of Hard Tissue Biology 27.1 (2018): 55-58.
David L. Wang, Chuanguang Xiao, Guofeng Fu, Xing Wang and Liang Li.
Identification of potential serum biomarkers for breast cancer using a functional proteomics technology. Biomarker Research (2017) 5:11. DOI 10.1186/s40364-017-0092-9
Zheng, H., et al. "
AlbuVoid™ coupled to on-bead digestion-tackling the challenges of serum proteomics." J Proteom Bioinformatics 8.9 (2015): 225.
Zheng, H., et al. Poster reprint first presented at 14th Human Proteome Organization World Congress (HUPO 2015), held on September 27 – 30, 2015 entitled “
New Proteomic Workflows Combine Albumin Depletion and On Bead Digestion, for Quantitative Cancer Serum”.
|
|
|
|
|
|
|
