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AlbuVoid™ - Albumin Depletion Plus Low Abundance Serum Protein Enrichment
- Albumin voids in flow-through >95%, with <30 minute bind/wash/elute protocol
- Low abundance enrichment equivalent or better than hexa-peptides or antibodies
- Disposable, cost-effective, no column regeneration or cross-contamination
- Mild elution maintains tertiary structure and simple transfer to secondary analysis
- The eluted fractions retain their enzymatic and biological activity
- Removes albumin from many species including human, mouse, sheep, bovine, goat, rat, calf, cell culture.
- The eluted fraction is compatible with LC-MS, activity based protein profiling and proteomic studies.
AlbuVoid™ is a albumin depletion reagent kit. It removes albumin from serum and plasma samples while concentrating low abundance, and/or low molecular weight proteins. The AlbuVoid™ protocol uses mild buffers; the protocol conditions are so gentle that native enzyme activity is retained in elution fractions. AlbuVoid™ considerably enhances resolution of proteins below 50 kD, a limitation of alternate enrichment protocols.
Click Here For AlbuVoid™ Product Sheet.
Click Here For AlbuVoid™ On Bead Digestion Protocol
AlbuVoid™ does not bind albumin. All other proteins(except albumin) in the sample binds to AlbuVoid™ and then you can elute off all the proteins minus the albumin. Resulting in low abundance serum protein enrichment. It is ideal for applications involving biomarker discovery, enzyme assays, toxicological studies for new drugs, protein profiling using SELDI analysis, protein array pixelation ,1D and 2D gel electrophoresis, LC/MS, and MALDI-TOF MS and cytokines research.
AlbuVoid™ derives from a silica-based library of individual mixed-mode polymeric ligands. The library was designed to facilitate weak binding of proteins, allowing for rapid elution from the matrix without any foreknowledge of the variety of proteins contained in the starting sample. Because of its specific binding properties, AlbuVoid™ depletes high abundance proteins in serum like albumin while improving the resolution of less abundant serum proteins.
Biotech Support Group has
AlbuVoid™ for albumin depletion plus low abundance serum protein enrichment. Both Albusorb™ and AlbuVoid™ are available in AlbuTrial™ kit™. AlbuTrial™ kit™ includes 1 gm of Albusorb™ and 5 Preps of AlbuVoid™ with respective buffers.
Product
|
Size
|
Total samples processed
|
Item No.
|
AlbuVoid™
|
10 Preps
|
10 x 200 µl samples
|
AVK-10
|
AlbuVoid™
|
50 Preps
|
50 x 200 µl samples
|
AVK-50
|
Kit includes:
Items Required
|
5 Prep
|
10 Prep
|
50 Prep
|
Reagent
|
AlbuVoid™
|
0.25 gram
|
0.5 gram
|
2.5 gram
|
Supplied
|
Binding Buffer AVBB, PH 6.0
|
4 ml
|
7 ml
|
35 ml
|
Supplied
|
Wash Buffer AVWB, PH 7.0
|
5 ml
|
10 ml
|
50 ml
|
Supplied
|
Elution Buffer AVEB, PH 9.8
|
2 ml
|
4 ml
|
20 ml
|
Supplied
|
SpinX Centrifuge tube filters
|
5
|
10
|
50
|
Supplied
|
Application Report
Haiyan Zheng1; Swapan Roy2; Devjit Roy MD2; Amenah Soherwardy1; Seema Rathman2; Matthew Kuruc2. 1Rutgers Center for Integrative Proteomics, Piscataway, NJ; 2Biotech Support Group LLC, Monmouth Junction, NJ. Stroma Liquid Biopsy – Pan-Cancer Dysregulation of the Serum Proteome.
Biotech Support Group reports on recent research, presented as a poster report and highlighted as a speaker presentation at the 14th Annual US HUPO Conference, held March 11-14, 2018 in Minneapolis, Minnesota USA. This research shows that through robust quantitative mass spectrometry workflows, systemic response to the presence of cancer can be profiled with protein biomarkers. These biomarkers fall within three common pathways which change in the bloodstream regardless of the primary tumor, stage, metastatic disease, or tumor burden. Five different cancers – lung, breast, pancreatic, lymph and ovary, were profiled along with normal/healthy individuals of approximate age and matched sex for comparison. “After several years of optimizing our AlbuVoid™ bead based enrichment methods and aggregating our data, I am very pleased that we can report what we believe are significant discoveries. Our selection of biomarkers offer key advantages as they are all highly observable, being of relative high abundance in the bloodstream. Furthermore, they are all highly differentiated – many severely, in the cancer population, and very stable in the normal/healthy population. Under the guidance of a physician, such dysregulated patterns may serve within the context of current ‘liquid biopsy’ platforms, forming an early indicator for cancer even before clinical evidence. These same patterns may be prognostic as well as offer individualized and precision therapy for a patient. We hope to refine our reporting methods with additional investigations in the future. For this, we are engaging partnerships with organizations to commercialize our proprietary discoveries, so ultimately we can improve cancer patient outcomes.”, states Dr. Swapan Roy, Ph.D., President and Founder of Biotech Support Group.
Synovial Fluid
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.
Biotech Support Group reports on a new research report which highlights the simplicity and efficiency of their proteomic sample preparation technology as an initial enrichment step for synovial fluid and serum analysis. Rheumatoid arthritis (RA) is characterized by chronic inflammation, hyperproliferation of synovial cells, and bone destruction. In this study, AlbuVoid™ was used to remove Albumin and enrich the low abundance proteomes from serum and synovial fluid. The article states “…albumin and globulin were eliminated by using an AlbuVoid Albumin Depletion Kit (Biotech Support, Monmouth Junction, NJ).” The investigation searched for RA-specific protein/peptide markers to facilitate early and sensitive diagnosis of RA using three different methods: a protein chip, 2D-gel, and western blotting. The study identified several RA-specific protein and peptide marker candidates. “We are very pleased to see that AlbuVoid™ provided the enrichment necessary to uncover biomarker candidates for Rheumatoid Arthritis. Using three different analytical reporting methods, the versatility of AlbuVoid™ enrichment from both synovial fluid and serum is demonstrated” states Swapan Roy, Ph.D., President and Founder of Biotech Support Group.
Serum
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.
Biotech Support Group reports on a new research report which highlights the simplicity and efficiency of their Albumin depletion sample preparation technology as an enrichment step for clinical research of human serum using LC-MS proteomic analysis.Proteins are essential for biomarker and disease discovery though the most abundant proteins often provide very limited clinically relevant data, as can be the case for albumin or immunoglobulins. In this work, the investigators focused on depleting albumin from human serum samples using an albumin depletion and low abundance protein enrichment kit – AlbuVoid™, which enabled the detection of several low-abundance proteins. By employing an optimized protocol, enriched proteins known as biomarkers for various diseases were identified. The article states “Protein capture was performed using AlbuVoid™ Kit on serum samples… For the LCMS analysis, elution was made directly on the beads with denaturant solution (30 μL of 8 M urea / 20 mM DTT / 100 mM Tris pH 8.5) before tryptic digestion. In comparison, methods using antibodies needed at least one-half day more. The albumin depletion method allowed to save precious time. ”. The authors concluded that the AlbuVoid™ depletion method proved to be faster and more cost-effective than antibody based methods, and could be helpful for biomarker enrichment and detection in medical research. “Throughout our product and methods development efforts, we have been especially focused on overcoming the challenges of clinical proteomics. So I am are very pleased to see that AlbuVoid™ was advantageously used for this purpose. Because we use a porous silica bead surface chemistry, the strong denaturing conditions studied here as the bead eluent. can provide a seamless workflow to FASP type digests. Consequently, the versatility of digest conditions, either FASP or on-bead digest - what we call BASP™, can offer many workflow and unique digest efficiency options. Collectively we continue to find new benefits for clinical discovery and targeted proteomics.” states Swapan Roy, Ph.D., President and Founder of Biotech Support Group.
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
Biotech Support Group contributed the chapter, “Methods to Monitor the Functional Subproteomes of SERPIN Protease Inhibitors,” Although SERPINs are found in a variety of functional on/off sub-forms, customarily they are observed and reported in aggregate. As a result, their influence on disease is missed. This chapter describes Albumin depletion products AlbuVoid™ & AlbuSorb™ and and demonstrates their utility to satisfy investigations of serum SERPINs at a functional level.
Poillet-Perez, Laura, et al. "Autophagy maintains tumour growth through circulating arginine." Nature (2018): 1.
Biotech Support Group reports on a new research report which highlights the simplicity and efficiency of their Albumin depletion sample preparation technology as an enrichment step for mouse serum LC-MS proteomic analysis. Autophagy is a cellular mechanism which captures intracellular components and delivers them to lysosomes, where they are degraded and recycled, helping cells survive during times of starvation. One in vivo model to study autophagy is whole-body deletion of the essential autophagy gene Atg7 in adult mice which causes a systemic metabolic defect that manifests as starvation intolerance. In order to measure the systemic proteomic response of such deletion in this study, AlbuVoid™ was chosen as one method to remove Albumin and enrich the low abundance proteomes from serum. The article states “…AlbuVoid (Biotech Support Group) was used to deplete albumin…”. This was done for the purpose of estimating differential protein abundances by LC-MS derived spectral counts. The serum proteomic investigation identified arginine-degrading enzyme arginase I (ARG1) in the circulation of Atg7-deficient hosts as one of several differentially identified proteins. Such evidence helped the authors’ conclude that defective autophagy in the host leads to the release of ARG1 from the liver and the degradation of circulating arginine, which is essential for tumour growth; thus identifying a metabolic vulnerability of cancer.“I are very pleased to see that AlbuVoid™ was used for the mouse serum LC-MS analysis. Although only one piece of this very interesting autophagy puzzle, it nevertheless helped provide evidence for a metabolic vulnerability in tumors” states Swapan Roy, Ph.D., President and Founder of Biotech Support Group.
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.
Biotech Support Group reports on a new research report which highlights the simplicity and efficiency of their proteomic sample preparation technology as an initial enrichment step for synovial fluid and serum analysis. Rheumatoid arthritis (RA) is characterized by chronic inflammation, hyperproliferation of synovial cells, and bone destruction. In this study, AlbuVoid™ was used to remove Albumin and enrich the low abundance proteomes from serum and synovial fluid. The article states “…albumin and globulin were eliminated by using an AlbuVoid Albumin Depletion Kit (Biotech Support, Monmouth Junction, NJ).” The investigation searched for RA-specific protein/peptide markers to facilitate early and sensitive diagnosis of RA using three different methods: a protein chip, 2D-gel, and western blotting. The study identified several RA-specific protein and peptide marker candidates. “We are very pleased to see that AlbuVoid™ provided the enrichment necessary to uncover biomarker candidates for Rheumatoid Arthritis. Using three different analytical reporting methods, the versatility of AlbuVoid™ enrichment from both synovial fluid and serum is demonstrated” states Swapan Roy, Ph.D., President and Founder of Biotech Support Group.
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
In this study, a functional proteomics technology was used to systematically monitor metabolic enzyme and protease activities from resolved serum proteins produced by a modified 2-D gel separation and subsequent Protein Elution Plate, a method collectively called PEP. AlbuVoid™ was used to remove Albumin and enrich the low abundance serum proteome. The article states “The most dramatic difference for enzyme activity detection in using the AlbuVoid™ for serum protein enrichment was demonstrated. … Compared with the direct serum proteinase measurement, both the levels and species of proteases were increased significantly in the enriched serum sample. …protease activity in the direct serum analysis suggested that the protease levels in the serum were below the detection threshold of protease activity…,and it is necessary to use AlbuVoid™ to enrich these low level proteases to bring them to a high enough level to be detected.” Both qualitative and quantitative differences in the metabolic enzyme and protease activity were detected between breast cancer patient and control group, providing excellent biomarker candidates for breast cancer diagnosis and drug development.
Swapan Roy, Matthew Kuruc. The Functional Subproteomes of Serpin Protease Inhibitors are Now Open for LC-MS Biomarker Discovery. MOJ Proteomics Bioinform 2016, 3(6): 00106
The authors consider that the conformational variants of the unique family of protease inhibitors annotated as SERPINs, are most often underrepresented in proteomic analyses. This limits understanding the complex regulation that this family of proteins presents to the networks within the protease web of interactions. Using bead-based separation provided by the NuGel™ family of proteomic enrichment products - notably AlbuVoid™ & AlbuSorb™, the authors demonstrate their utility to satisfy investigations of serum SERPINs. The authors also suggest their use to develop functional profiles of the SERPIN Proteoform, and how those can establish relationships to disease phenotypes, gene mutations, and deregulated mechanisms.
Featured Application - Functional Proteomics Sun, Zhenyu, Xiaofeng Chen, Gan Wang, Liang Li, Guofeng Fu, Matthew Kuruc, and Xing Wang. "Identification of functional metabolic biomarkers from lung cancer patient serum using PEP technology." Biomarker Research 4, no. 1 (2016): 1.
In brief, the article’s authors report a on a functional proteomics top-down method to systematically monitor metabolic enzyme activities in resolved serum proteins produced by a modified 2-D gel separation and subsequent Protein Elution Plate, a method collectively called PEP. The article states “Since most of the functional proteins or enzymes exist at relatively low level in the human serum and there is a limited loading capacity on the 2-DE gel, it is important to enrich the low abundance proteins before 2-DE and PEP analysis. AlbuVoid™ (Biotech Support Group, Monmouth Junction NJ) has been shown to effectively enrich low abundance serum proteins while depleting the Albumin…whereby more functional features were observed with AlbuVoid™ than without…”. The study identified several potential functional enzyme biomarkers from lung cancer patient serum and evidence that the methods provide an alternative and complementary approach to sequence annotation for the discovery of biomarkers in human diseases.
“We are pleased that AlbuVoid™ integrates so well with the PEP platform. Maintaining the integrity of the functional features of proteins is a key feature in all of our enrichment products, as the landscape of protein conformations is highly variable; each conformation contributing to its own unique functional activity. Furthermore, the PEP technology supports great resolving power in functional proteomic studies”, states Swapan Roy, Ph.D., President and Founder of Biotech Support Group.
Cancer Proteomics Application Report
Poster Reprint First Presented At AACR Annual Meeting 2016 Conference, Held April 17-20, 2016 New Orleans, LA USA. The Commonality of the Cancer Serum Proteome Phenotype as analyzed by LC-MS/MS, and Its Application to Monitor Dysregulated Wellness Haiyan Zheng1; Caifeng Zhao1; Swapan Roy2; Devjit Roy MD; Amenah Soherwardy2; Ravish Amin2; Matthew Kuruc21Rutgers Center for Integrative Proteomics, Piscataway, NJ; 2Biotech Support Group LLC, Monmouth Junction, NJ
Princeton based Biotech Support Group announced today that it has submitted provisional patents disclosing its data on many common proteins dysregulated in cancer. This breakthrough research shows that most of these biomarkers are believed to be dysregulated
regardless of the primary tumor, stage of progression or tumor burden. The study which collaborated with the Rutgers Center for Integrative Proteomics shows the most distinguishable pattern of early dysregulation observable as a cancer serum phenotype to date. This forms the basis of a new commercialization plan featuring:
Variant sub-populations (also known as proteoforms) of a common blood protein – Alpha-1-Antitrypsin, with functional reporting features severely distinguishable between cancer patients and normal/healthy individuals
A measurable serum cancer profile that can be modeled with categorical biomarker proteins taken from inflammation, blood coagulation, tissue remodeling, and glycolysis, as well as new markers of unknown function.
Dr.Swapan Roy; Amenah Soherwardy; Ravish Amin; Matthew Kuruc Biotech Support Group LLC, Monmouth Junction, NJ - Application Report. AlbuSorb™ Product Extension combines Albumin and Immunoglobulin Depletion in a Consumable Format
The NuGel™ platform chemistry, a porous polymer coated (ionic, polymeric, aliphatic) silica library of bead architecture allows liquid chromatography mass spectometry experiments. Albusorb™ from Biotech Support Group depletes albumin. Proteomic enrichment of low abundance proteins from beads and high abundance serum protein depletion may require albumin and immunoglobulin removal. Samples can be human serum or mouse plasma. To obtain LC-MS spectra counts various parameters such as fixed modification (carbamide methylation or cysteine and SDS with Tris) and variable modification (deamidation on Asparagine oxidation) of samples can be set. Some experiments on globin depletion can require protein A combination protocols. A database of serum proteins can be quantified on cancer sera phenotype and normal healthy patients. The sub proteome functional analysis can be applicable to downstream experiments such as antibody arrays, ELISA, LC-MS, 1&2 DE, Western blot etc.
Haiyan Zheng1; Caifeng Zhao1; Swapan Roy2; Devjit Roy MD; Amenah Soherwardy2; Ravish Amin2; Matthew Kuruc2
1Rutgers Center for Integrative Proteomics, Piscataway, NJ; 2Biotech Support Group LLC, Monmouth Junction, NJ. Application Report - The Comparison of the Serum Proteome in Individuals with Cancers versus those without Cancer, and its application to Wellness
Cancer Biomarker and Wellness protein discovery
Biomarker discovery from genomic analysis of singlular biomarkers or protein panels can be enhanced by doing albumin depletion experiments and on bead digestion.
A wellness proteome strategy of over 200 spectrally quantified proteins from healthy/normal or disease cancerous sera of pancreatic, breast, lung patients are obtained.
Individuals from pancreatic, breast and lung cancer sera are analyzed by adding a binding buffer. The bead retains serum proteome and albumin is depleted. Subsequently the serum proteins on bead are analyzed by wash, reduction, alkylation, trypsin digestion and are labelled. Spectra from liquid chromatography mass spectrometry (LC-MS/MS) runs and proteins analyzed by Xtandem with protein filters are obtained.
An example of a protein is the SERPIN family protease inhibitor. It allows validating quantitative proteomics data, discoverying newly annotated serum proteome.
Protein
ID
|
Protein
Description
|
Plasma
Proteome Annotation
(approx.
conc.)
|
TMT
Report Ratio
Threshold
|
#
Individuals Threshold/All
|
Cancer
Types
|
Evidence
in Cancer Sera by (Others)
|
Comments
|
ITIH3/4
(at least one subunit
chain)
|
Inter alpha trypsin
inhibitor, heavy chain ¾
|
40 µg/ml
|
>
1.5
(up-regulated)
|
7/13
|
Pancreatic,
Breast,
Lung
All Stages
|
Up-regulated
(3)(4)(5)(6)
|
4 Sub-unit protease
inhibitors, suspected role in inflammation
|
APOA1
|
Apolipoprotein A I
|
1 mg/ml
|
<
0.7
(down-regulated)
|
9/13
|
Pancreatic,
Breast,
Lung
All Stages
|
Down-regulated (7)
|
major protein
component of high density lipoprotein (HDL)
|
APOC3
|
Apolipoprotein C III
|
100-200 µg/ml
|
<
0.7
(down-regulated)
|
5/13
With
APOA1
11/13
|
Pancreatic,
Breast,
Lung
4/5 Stage 1
|
Up-regulated
(8)
|
a
component of very low density lipoprotein (VLDL)
Our results conflict
with other reports, maybe APOC3 reports as cyclical up & down
dysregulation
|
CRP
|
C-reactive protein
|
1-2 µg/ml
|
>
1.5
(up-regulated)
|
7/11
|
Pancreatic,
Breast,
Lung
All Stages
|
Up-regulated (9)
|
Acute phase response
protein, may scavenge nuclear material released from damaged
circulating cells
|
PF4
|
Platelet Factor 4
|
5-10 ng/ml
|
>
1.5
>
2.0
(up-regulated)
|
13/13
9/13
|
Pancreatic,
Breast,
Lung
All Stages
|
Up-regulated (8
)(9)(10)
|
promotes blood
coagulation, may play role in wound repair and inflammation
|
PPBP
|
Beta
thromboglobulin
aka CTAPIII/NAP-2
|
5-10 µg/ml
|
>
1.5
>
2.0
(up-regulated)
|
13/13
9/13
|
Pancreatic,
Breast,
Lung
All Stages
|
Up-regulated (1)
|
Platelet
activation
Elevated levels
predated lung cancer diagnosis by 29 months, with cyclical levels
upon surgical resection and recurrent disease (1).
|
CLU
|
Clusterin
|
100 µg/ml
|
>
1.5
(up-regulated)
|
12/13
|
Pancreatic,
Breast,
Lung
All Stages
|
Up
& down-regulated
(11)
|
Multiple
isoforms, associated with clearance of cellular debris and tissue
remodeling.
Up & down
regulation have been described for many cancers, may be isoform
specific (11)
|
Read the application report to view more proteins from cancer patient samples.
These proteins function on tissue remodeling, wound repair, inflammation, transport of polymeric immunoglobulins, platlet activation, glycolysis regulation, cell viability, growth factor inhibition, MAP kinase activation, metalloproteinases, acute phase markers, neural development and as thyroid hormone carriers, components of VLDL and HDL, multiple isomers etc.
Peptide level annotation and regulation of protease inhibition are examples of research applications of the wellness proteins. Variant subproteomes in proteomic workflows.
This research suggests how find measurable serum cancer phenotypes. Searching and discovering new conformational and isoform variants with high abundance proteins from SERPIN, clusterin, immunoglobulin requires albumin depletion.
Such research progresses biomarker discovery (inflammatory, tissue remodeling, glycolysis, coagulation etc) and minimizes protelytic noise and has less digestive time.
Xing Wang, Michael Davies, Swapan Roy and Matthew Kuruc. Bead Based Proteome Enrichment Enhances Features of the Protein Elution Plate (PEP) for Functional Proteomic Profiling . Proteomes 2015, 3(4), 454-466 .
doi:
10.3390/proteomes3040454
In this short case study, the enrichment of select sub-populations of proteins is beneficial to systematically analyze protein functions of a whole enzyme family from an entire proteome. AlbuVoid™ was used to remove Albumin and enrich the low abundance proteome, noting that distinguishable activity features are presented from the lung cancer vs. the normal sera. KinaSorb™ was used to enrich for both a narrow spectrum substrate profile—Hexokinase activity, and a broad-spectrum protein kinase activity. The number of observable features was consistent with such narrow and broad-spectrum activities. AlbuVoid™ enrichment and PEP processing proved suitable for profiling the functional activities of Hexokinase, Protease and Alkaline Phosphatase. These enzyme feature profiles are indicative of the functional diversity that can be generated, annotated and compared within and between sample phenotypes, using the combined methods.
Zheng et al.
AlbuVoid™ Coupled to On-Bead Digestion - Tackling the Challenges of Serum Proteomics. J Proteomics Bioinform 2015, 8:9
Abstract
For cancer research, serum and plasma are especially attractive sample types as collection of blood is common, simple and only minimally invasive. Yet serum samples can offer unique challenges in LC-MS proteomic analyses. The two biggest challenges being: 1) the high abundance of Albumin accounting for about 50% of the total protein mass and, 2) proteolytic resistance, in large part due to substantial amount of glycoprotein, a modification that manifests proteolytic resistance. In this short report, we describe new methods using a surface/bead based product, AlbuVoid™, which depletes Albumin through a negative selection or voidance strategy, retaining the vast amount of the remaining serum proteome on the bead. We then combine this novel enrichment, with a direct and seamless integration with Trypsin digestion, a method conventionally referred to as on-bead digestion. We evaluated the digestion time as a parameter to identify whether different sub-populations of peptides and proteins can be observed by LC-MS analyses. Using 2 different allotted digestion times - 4 hours, and overnight, each with a singular 3 hour gradient LC-MS run, between 400-500 total proteins were observed for both human and rat sera, with overlapping and distinct sub-populations observable at each digest time. These results support that the described methods gain efficiencies over other high abundance depletion and in-solution digestion workflows. We solicit that such workflows will minimize many of the inconsistencies of proteolytic hydrolysis for both discovery and quantitative serum proteomic applications.
Genetic Engineering News Omics Tutorial Describes Biotech Support Group and ArrayBridge Joint Functional Proteomic Data
A Genetic Engineering News article describes a joint collaboration with ArrayBridge (St. Louis, MO). The article describes the combination of first low abundance protein enrichment/high abundance protein depletion with the Biotech Support Group product – AlbuVoid™, followed by a modified 2-dimensional electrophoretic separation, and transfer via the ArrayBridge PEP plate into microplates. From there, the resolved functionally active proteins were measured and characterized.
The citation is:
Xing Wang, Ph.D., Zhenyu Sun, M.D., Xiaofeng Chen, M.D., Xiong Su, Ph.D., Gan Wang, Ph.D., Matthew Kuruc. Genetic Engineering News Jan 1, 2015 (Vol. 35, No. 1) OMICS Tutorial Discovery of Functional Serum Biomarkers, Exploring Cancer's Signature in the Sensitive Functional Domain of the Human Proteome
http://www.genengnews.com/gen-articles/discovery-of-functional-serum-bio...
Plasma
Espes, Daniel, Joey Lau, and Per-Ola Carlsson. "Increased circulating levels of betatrophin in individuals with long-standing type 1 diabetes." Diabetologia(2013): 1-4.
Authors Espes et al published an article in the journal Diabetologia which discovered an increase in hormone betatrophin in type 1 diabetics as compared to health individuals. Betatrophin causes an increase of pancreatic β cell replication and regulates glucose levels. Found in liver and adipose tissue, the hormone also increases β cell mass expansion. Analysis of betatrophin by mass spectrometry (MS) or gel electrophoresis could require enriching the protein mixture and depleting high abundance proteins to display less abundant proteins. The article titled, “Increased circulating levels of betatrophin in individuals with long-standing type 1 diabetes” utilized AlbuVoid™’s albumin depletion protocol for removing albumin from plasma samples. The article quotes AlbuVoid™ from Biotech Support Group, “Plasma samples were depleted of albumin using AlbuVoid Albumin Depletion Kit (Biotech Support Group, Monmouth Junction, NJ, USA). Data were normalized for total protein content.” Betatrophin increases proliferation of beta cell and this study identified double the concentration of betatrophin as measured by western immunoblot using a betatrophin primary antibody in type 1 diabetics. Moreover factors such as age in healthy controls displayed a direct relationship with increase in betatrophin whereas triacylglycerol, LDL-cholesterol, HDL-cholesterol levels and insulin were not affected. Increasing concentrations of betatrophin did not prevent against the loss of C-peptide suggesting type 1 diabetes on betatropin treatment would benefit from combination treatment. The authors concluded, “An intervention in patients with type 1 diabetes with betatrophin treatment might require supraphysiological dosing as well as combination with immune regulatory treatment.”
Cell Culture
Narain, Niven Rajin, Rangaprasad Sarangarajan, Vivek K. Vishnudas, and Michael Andrew Kiebish. "USE OF MARKERS IN THE IDENTIFICATION OF CARDIOTOXIC AGENTS AND IN THE DIAGNOSIS AND MONITORING OF CARDIOMYOPATHY AND CARDIOVASCULAR DISEASE." U.S. Patent 20,140,100,128, issued April 10, 2014. Secretome sample preparation involves preparing a medium of cells cultured in media not containing serum which is concentrated, reduced, alkylated, desalted and digested. Another medium of cells is cultured in serum containing medium, reduced using columns and reconstituted using PBS. Serum albumin is depleted from these samples using Biotech Support Group's AlbuVoid™ column.
The patent quotes "In one embodiment, the cells can be cultured in serum containing medium: The volume of the medium can be reduced using 3k MWCO Vivaspin columns (GE Healthcare Life Sciences), then can be reconstituted with 1×PBS (Invitrogen®). Serum albumin can be depleted from all samples using AlbuVoid column (Biotech Support Group, LLC) following the manufacturer's instructions with the modifications of buffer-exchange to optimize for condition medium application."
Patents
Patent Application Cites AlbuVoid™ For Secretome Sample Preparation & Neuroscience Research
Narain, Niven Rajin, and Paula Patricia Narain. COMPOSITIONS AND METHODS FOR DIAGNOSIS AND TREATMENT OF PERVASIVE DEVELOPMENTAL DISORDER United States Patent Application 20150023949, Pub. Date 01/22/2015.
The patent provides albumin sample preparation research application to track modulators of disease processes and experiment data on metabolomics, transcriptomics, single nucleotide polymorphisms using sample preparation and an artificial intelligence based informatics platform. Serum containing media are reduced using columns, reconstituted with buffers and serum albumin is depleted using Biotech Support Group's AlbuVoid™.
Narain, Niven Rajin, Rangaprasad Sarangarajan, and Vivek K. Vishnudas. "INTERROGATORY CELL-BASED ASSAYS AND USES THEREOF." U.S. Patent No. 20,120,258,874. 11 Oct. 2012.
Serum albumin depletion for secretome sample preparation using AlbuVoid™ while developing a cellular modeling system. The patient cites AlbuVoid™ from Biotech Support Group “In one embodiment, the cells can be cultured in serum containing medium: The volume of the medium can be reduced using 3k MWCO Vivaspin columns (GE Healthcare Life Sciences), then can be reconstituted withlxPBS (Invitrogen). Serum albumin can be depleted from all samples using AlbuVoid column (Biotech Support Group, LLC) following the manufacturer's instructions with the modifications of buffer-exchange to optimize for condition medium application.”
Patent - Serum Containing Medium Narain, Niven Rajin, Rangaprasad Sarangarajan, and Vivek K. Vishnudas . "Interrogatory cell-based assays for identifying drug-induced toxicity markers." U.S. Patent Application No. 16/180,446.
Biotech
Support Group reports on a new patent application which highlights
the simplicity and efficiency of their Albumin depletion sample
preparation technology as an enrichment step for serum containing
medium used in cell culture for LC-MS proteomic analysis. The
lack of reliable tools that can help with predicting toxicity early
in drug development is partly to blame for a 90% attrition of
potential compounds entering clinical development, 30% of which is
owing to poor clinical safety. Cardiotoxicity particularly is a
leading cause of drug withdrawal. The invention described is useful
for identifying markers associated with drug-induced toxicity. The
patent application states “In
one embodiment, the cells can be cultured in serum containing medium:
The volume of the medium can be reduced using 3 k MWCO Vivaspin
columns (GE Healthcare Life Sciences), then can be reconstituted with
1×PBS (Invitrogen). Serum albumin can be depleted from all samples
using AlbuVoid column (Biotech Support Group, LLC) following the
manufacturer's instructions with the modifications of buffer-exchange
to optimize for condition medium application.” The patent
description includes a novel, collaborative utilization of network
biology, genomic, proteomic, metabolomic, transcriptomic, and
bioinformatics tools and methodologies, which, when combined, may be
used to study any biological system of interest, such as obtaining
insight into the molecular mechanisms associated with or causal for
drug-induced toxicity. “I
are very pleased to see that AlbuVoid™ was referenced for the
proteomic analysis of cell culture. As we do not use immunoaffinity,
separations are efficient across all species” states Swapan Roy,
Ph.D., President and Founder of Biotech Support Group.
Poster
Improved proteomic enrichment and workflow strategies
US HUPO 2014, Poster 089
Proteomic sample preparation technology for depleting hemoglobin, and enriching the low abundance proteome from human erythrocyte lysates and determination of the position and nature of human protein N termini in different tissues and disease states. Biotech Support Group has developed a series of enrichment reagents that offer common features and key advantages. After separations, the sub-proteomes retain their structural and functional integrity. All the products are consumable and economical; not being derived from biological sources. We report here applications for these products to specifically deplete albumin by a voidance strategy - called AlbuVoid™, which consequently enriches for the low abundance serum protein content. Such a strategy compares favorably to high abundance immuno-depletion (Agilent column) methods. Also, we continue to advance simplified workflows that utilize on-bead digestion for LC-MS. Together, these methods create efficiencies necessary to support high throughput investigations in all areas of discovery, targeted, functional and chemical proteomics.
New On-Bead Digestion Protocols Improve LC-MS Workflows Of Albumin Depleted Samples US HUPO 2013 Translational Proteomics
Biotech Support Group LLC, reports on a new technical poster which describes the use of on-bead digestion protocols as a suitable method to generate peptides for LC-MS/MS analysis, after albumin depletion on its AlbuVoid™ product. Entitled “New On-Bead Digestion Protocols Improve LC-MS Workflows Of Albumin Depleted Samples”, it was presented at the US HUPO Conference in Baltimore, MD, March 10-11, 2013. It describes by several performance metrics, that the ‘on-bead’ digestion protocol is comparable to the product’s standard elution protocol. On-bead protocols provide advantaged speed, simplicity and reduced potential for keratin contamination. The poster also reports on the efficiency of AlbuVoid™ as a simple consumable product for albumin depletion, with 20-30% more protein/peptide identifications achievable relative to untreated plasma, under identical operating and informatic parameters. “With greater interest in the proteomics community for better workflows and performance for LC-MS analyses, direct proteolytic digestion of the bound protein content - commonly called on-bead digestion, has many desirable workflow advantages. With this report on AlbuVoid™, we have now demonstrated that the surface chemistry platform from which it was derived, our novel passivated porous silica –NuGel™, offers the rigidity and low water content ideal for protein concentration and proteolytic digestion”, states Swapan Roy, Ph.D, President & Founder of Biotech Support Group. “We envision designing new strategies for high abundance depletion that can utilize on-bead digestion workflows for highly complex bead-bound proteomes along with many chemical proteomic applications.”
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1; Swapan Roy2; Devjit Roy MD2; Amenah Soherwardy1; Seema Rathman2; Matthew Kuruc2. 1Rutgers Center for Integrative Proteomics, Piscataway, NJ; 2Biotech Support Group LLC, Monmouth Junction, NJ.
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Poster Reprint First Presented At AACR Annual Meeting 2016 Conference, Held April 17-20, 2016 New Orleans, LA USA. The Commonality of the Cancer Serum Proteome Phenotype as analyzed by LC-MS/MS, and Its Application to Monitor Dysregulated Wellness Haiyan Zheng1; Caifeng Zhao1; Swapan Roy2; Devjit Roy MD; Amenah Soherwardy2; Ravish Amin2; Matthew Kuruc21Rutgers Center for Integrative Proteomics, Piscataway, NJ; 2Biotech Support Group LLC, Monmouth Junction, NJ
Xing Wang, Michael Davies, Swapan Roy and Matthew Kuruc. Bead Based Proteome Enrichment Enhances Features of the Protein Elution Plate (PEP) for Functional Proteomic Profiling . Proteomes 2015, 3(4), 454-466 . doi: 10.3390/proteomes3040454
Haiyan Zheng, Caifeng Zhao, Meiqian Qian, Swapan Roy, Absari Arpa, Amenah Soherwardy and Matthew Kuruc. AlbuVoid™ Coupled to On-Bead Digestion - Tackling the Challenges of Serum Proteomics. J Proteomics Bioinform 2015, 8:9.
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AlbuVoid™ LC-MS On-Bead – Differential Expression of Cancer Sera Proteins Using Quantitative (iTRAQ) Proteomics
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On-Bead Digestion Protocols For LC-MS Proteomic Workflows
New on-bead digestion for LC-MS applications for proteomic studies
US HUPO 2014. Frontiers in Proteomics: Advancing Biology through Technology and Computation.
AlbuVoid™ abstract entitled "Improved proteomic enrichment and workflow strategies", poster board 089 presented at US HUPO 2014
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