Sample Prep – Liquid Biopsy
Proteomic Sample Prep for
- Urine
- Blood/Serum/Plasma
- Saliva/Sputum
- Synovial Fluid
- BALF
- Exosomes
With the emergence of liquid biopsy biomarkers, BSG offers two complementary technologies for proteomic researchers to selectively bind or enrich, in order to achieve the best results. By using one of these two solutions, researchers will be able to improve efficiency and outcomes regardless of the analytical platform. This will be especially beneficial for discovery investigations requiring cost effective and efficient sample prep methods essential for expanding proteomics into routine healthcare.
UPCK™ Kit
Urine Protein Enrichment & Concentration
- Linearly scaleable, unlike ultrafiltration
- Alternative to solvent/ alcohol precipitation
- On-bead digestion protocols
- <60 min. bind, wash, elute protocol
- Applicable to
>1 & 2 DE
>LC-MS
>microarrays - The eluted fractions retain their enzymatic and biological activity
AlbuVoid™
Selectively Voids Albumin, Binds Low Abundance Proteome
- Albumin voids in flow through, >95%
- <30 minute protocol
- Low abundance enrichment equivalent or better than hexapeptides or antibodies
- On-bead digestion protocols, efficient LC-MS workflows
- Disposable, cost-effective, no column regeneration or cross-contamination
2DE analysis of AlbuVoid™ treated sheep serum. Samples were reduced, alkylated and total protein normalized. The circled regions indicate the albumin zone. 1: Control. 2: AlbuVoid™ Eluate. Bottom: AlbuVoid™ Eluate silver stain.
Notable Reference
For serum & synovial fluid
Identification of Specific Protein Markers of Rheumatoid Arthritis in Synovial Fluid and Serum
AlbuVoid™ PLUS
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.
20µg of protein loaded on to 10% tris bis gel
A - Serum alone – shows IgG and Albumin regions
B - NuGel™ ProteinA treated serum – shows IgG depletion
C - AlbuVoid™ treated serum – shows Albumin depletion
D - AlbuVoid™ PLUS treated serum – shows IgG and Albumin depletion
AlbuSorb™
Selectively Binds Albumin
- Removes serum albumin >90%
- Economical small ligand surface architecture (not dye-based), bio-affinity performance
- Species agnostic
AlbuSorb™ PLUS
Selectively Binds Albumin & Immunoglobulin
- > 85% Albumin, 85% IgG depleted from 25 μl serum
Quantitative Bias Comparison between
AlbuSorb™ & AlbuSorb™ PLUS
| AlbuSorb™ | AlbuSorb™ PLUS | |
|---|---|---|
| Total IG IDs 2+ Spectral Cts | 80 | 58 |
| Total IG % of Spectral Count | 24% | 9% |
| Total Albumin % of Spectral Count | 12% | 15% |
Notable References
For Urine exosomes
For Synovial Fluid
PRELP protein inhibits the formation of the complement membrane attack complex
For Serum exosomes
HemoVoid™
Hemoglobin Depletion For Erythrocyte Proteomics
- Hemoglobin voids in flow-through, applicable to red cells, heavily hemolyzed serum, whole blood and dried blood spot (DBS) card
- Low abundance protein and enzyme enrichment
- Consumable, cost-effective
- Mild elution maintains native structure with retained enzymatic, functional and bioactivities
- Compatible with LC-MS, activity-probe profiling and virtually all proteomic analyses
- Species agnostic
2DE Comparison.
Red circles indicate the Hemoglobin subunits region. The HemoVoid™ eluate (bottom) has been severely depleted of Hemoglobin. The remainder of the red cell proteins are substantially enriched (visualized) and are better resolved in the HemoVoid™ eluate. Many more proteins are detectable after HemoVoid™ treatment with extensive proteome coverage across both dimensions.
Notable References
HemoVoid™ Blood Card Kit
The HemoVoid™ Blood Card kit substantially reduces hemoglobin interference from dried blood spot protein analytes
View Details
HemogloBind™
and
NuGel™ HemogloBind™
Removes Hemoglobin Interference
- Highly specific for hemoglobin binding
- depletion from hemolyzed serum, dura, BALF, and whole blood
- supports biomarker quantitative analysis
Notable References
For Bronchoalveolar Lavage Fluid (BALF)
Proteomic Studies in Acute Hypoxic Respiratory Failure.
For Dried Blood Spot (DBS)/Whole Blood
A simple assay for glutathione in whole blood
For hemolyzed serum/plasma exosomes
For hemolyzed serum/plasma
Evaluation of HemogloBind™ treatment for preparation of samples for cholinesterase analysis.
HemogloBind™ Blood Card Kitt
Hemoglobin Depletion and Protein Enrichment From Dried Blood Spots
- Dried blood spots are useful for low volume analyses, and simple collection and transport
- Protocols suitable for inexpensive whole blood card systems, no need for cell separation
- Hemoglobin binding >90%, with 30-45 minute spin-filter format
- Protocols based on ≤10 µl whole blood applied, but suspension format is flexible to most volumes
- Blood proteins and enzymes are enriched for biomarker and proteomic investigations.
- Removes hemoglobin from diverse species including human, sheep, bovine, goat, rat, mouse, etc.
- High throughput easicly scalable.
Cleanascite™
Lipid Adsorption & Clarification
- Effectively replaces chlorinated/fluorinated hydrocarbons (eg. freon)
- Workflows for antibodies, proteins, nucleic acids, proteoglycans, and most serum analytes
- A high binding capacity for lipids with minimal cross-reactivity with proteins and nucleic acids
Cleanascite™ is supplied as a suspension reagent. Simply add, mix and centrifuge in a 10 minute protocol.
Notable References
Plasma/Serum Protein Biomarkers
For diabetes biomarkers, Cleanascite™ is shown both to improve LC-MS measurements, and validated in accordance with CLIA ’88 guidelines. { doi: 10.1016/j.cca.2016.01.019 }
For saliva/sputum/tracheal swab
A nanobeads amplified QCM immunosensor for the detection of avian influenza virus H5N1
Isolation of M. tuberculosis RNA from Sputum
NuGel™ PBA
and
NuGel™ PBA Kit
Glycoprotein Enrichment Using Phenyl Boronic Acid
- Enriches heterogeneous sets of glycoprotein’s, N-linked & O-linked
- Consumable, no column regeneration
- Species and tissue agnostic
- Sorbitol elution; compatible with functional assays, electrophoresis and LC-MS
- Binds biomolecules containing 1,2 cis-diol groups
- Chemically derived, ideal for glyco-proteomic applications
- NuGel™ polymer coating, porous silica based
- Supplied as bead only (dry powder) or as kit (includes all binding and elution buffers)
| Sample Type | %Glycoprotein (Sorbitol Elution) |
|---|---|
| Mouse Plasma | 33 |
| Rat Serum | 44 |
| Sheep Serum | 18 |
| Bovine Serum | 40 |
| Bovine Brain Homogenate | 9 |
Different heterogeneous sets of glycoproteins are observed from 4 different mammalian
Gel Key:
- A: Mouse Plasma Eluate
- B: Sheep Serum Eluate
- C: Bovine Serum Eluate
- D: Rat Serum Eluate
Our BSG Advantage
Consumable
Cost-effective, not derived from biologicals
- No specialized instruments or HPLC
- Economical surface chemistries, not derived from biologicals
- No regeneration, so no prep to prep variability
- Simple, fast microfuge bind/wash/elute protocols
On-Bead Digestion
Efficient workflows, quality LC-MS/MS data
- Simple, reproducible workflows
- Equivalent or better than in-solution digestion
- Seamless to LC-MS, no desalting or C18 separations
- Unique proteolytic efficiencies
Enrichment/Depletion
Diverse strategies, species agnostic
- Products support strategies for both enrichment of low abundance proteomes, or depletion of high abundance proteins
- Species agnostic not derived from biologicals
Functional Integrity
Maintained throughout all separations
- Mild buffer conditions maintains native structure with retained enzymatic, functional & bio-activities
- Supports enzyme biomarker assays
- Functional & Chemical Proteomics
- Structural & activity-probe Proteomics
- Top-down & ArrayBridge PEP Proteomics