Key References

This study aimed to report the post-translational identity, biological activity and concentration of monomethyl and dimethylarginine-modified proteins by using GC-MS and LC-MS/MS approaches. The article states “we included in our method the use of HemoVoid™ … to improve the SDS-PAGE separation of proteins for further processing. HemoVoid™ …allowed removal of erythrocytic hemoglobin to a large extent. … removal of hemoglobin by this technique enabled an effective separation by SDS-PAGE and isolation of bands, presumably by avoiding overloading of the gels by hemoglobin.”.

Bollenbach, Alexander, et al. "GC-MS and LC-MS/MS pilot studies on the guanidine (NG)-dimethylation in native, asymmetrically and symmetrically NG-dimethylated arginine-vasopressin peptides and proteins in human red blood cells." Journal of Chromatography B (2020): 122024.

The article describes the advantage of HemoVoid™ in detection of low abundance proteins when comparing their amounts (in percent) between with four other extraction conditions. Most peptides, following HemoVoid™ extraction, “showed ion abundances ranging between 1.00E+5 and 1.00E+6 (31%). In comparison to this, fewer peptides (10–23%) were within this range following extraction with all other protocols”. With respect to potential biomarkers for Parkinson’s Disease, the article states “For example, PRDX6 accounts for 0.4% of the total ion abundance after DOC (deoxycholate) extraction, whereas following HV (HemoVoid™) extraction, this increases to 8%, a 20-fold enrichment”. The authors conclude that the HemoVoid™ method significantly reduces the concentration of hemoglobin, resulting in an increased signal-to noise of the remaining red cell proteins. The article describes methods to digest the HemoVoid™ bead-bound proteome, greatly simplifying the workflow for LC-MS/MS analysis.

Klatt, Stephan, et al. " Optimizing red blood cell protein extraction for biomarker quantitation with mass spectrometry." Analytical and Bioanalytical Chemistry (2020): 1-14.

Key References

HemogloBind ™ Cited in Comparison Proteomics Study of Covid-19 Patients in Peripheral Blood Derived Mononuclear Cells (PBMCs) – The Barometer of the Immune System

HemogloBind ™ sample preparation technology provides deep and quantitative LC-MS proteome and phosphoproteome analysis. This study provides a comparison of SARS-CoV-2 positive ICU patients with age- and sex-matched SARS-CoV-2 negative ICU patients and healthy individuals, using peripheral blood derived mononuclear cells (PBMCs).

Kaneko, Tomonori, et al. "System-wide hematopoietic and immune signaling aberrations in COVID-19 revealed by deep proteome and phosphoproteome analysis." Research Square preprint (2021).

A research article describes the simplicity and efficiency of BSG's hemoglobin depletion technology for 2X improvement of proteomic information obtained from bronchoalveolar lavage fluid (BALF).

Maneesh Bhargava M. et al."Proteome Profiling in Lung Injury after Hematopoietic Stem Cell Transplantation". Biol Blood Marrow Transplant 22 (2016) 1383-1390. http://www.bbmt.org/article/S1083-8791(16)30060-X/fulltext

The cellular thermal shift assay (CETSA) enables proteome-wide target screening for unmodified antimalarial compounds with undetermined mechanisms of action, providing quantitative evidence about direct drug–protein interactions. The article states “The intact-cell CETSA protocol features a HemogloBind- based sample processing step, which provides a relatively fast, reliable and inexpensive method to deplete >90% of hemoglobin from processed intact-cell samples. As a result, it leads to a 40-50% increase in the number of peptide spectrum matches (PSMs) for P. falciparum and non-hemoglobin human proteins.”. The citation is: Dziekan, Jerzy Michal, et al. "Cellular thermal shift assay for the identification of drug–target interactions in the Plasmodium falciparum proteome." Nature Protocols (2020): 1-41.

Key References

Biofluids

Farina, Annarita. " Pre-fractionation of Noncirculating Biological Fluids to Improve Discovery of Clinically Relevant Protein Biomarkers." Proteomics for Biomarker Discovery. Humana Press, New York, NY, 2019. 23-37.
For proteomic biomarker discovery, it is necessary to bridge the gap between basic and applied research by complying with clinical requirements. This chapter provides key suggestions for improving the discovery of clinically relevant protein biomarkers from body fluids. The chapter states : ”If the elimination of lipids…is necessary, the sample can by treated with lipid removal (Cleanascite™)…

Plasma/Serum Protein Biomarkers

The authors aimed at simultaneously measuring intact insulin and proinsulin derived C-peptide, to help predict development of diabetes mellitus, as well as in differential diagnosis in cases of hypoglycemia. 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 }

Vaccine Development

To evaluate immunogenic response to a vaccine candidate, it is necessary to measure the antibodies from sera; a sample with a diverse lipid profile. In this citation, Cleanascite™ was used in a toxin neutralizing assay to evaluate the influence of cholesterol dependency, on a candidate protein pneumococcal vaccine. { doi: 10.1016/j.vaccine.2012.11.005 }

Ascites Monoclonal Antibodies

The researchers determined the role of complement on MAb-mediated protection for four mice Ig subclasses. After centrifugation of ascetic fluid, Cleanascite™ protocol was implemented to remove lipids. { doi: 10.1128/IAI.70.5.2598-2604.2002 }

Cellular Response Applications

The applications and references for the many diverse investigations using Cleanascite™ upstream of cell response measurements are described. { Cleanascite Cell Response Reference Applications }

Tracheal Swab Samples

Li D, Wang J, Wang R, Li Y. A nanobeads amplified QCM immunosensor for the detection of avian influenza virus H5N1, Biosensors and Bioelectronics.2011;26(S10):4146-4154 Fu LM, Shinnick TM. Genome-wide exploration of the drug action of capreomycin on Mycobacterium tuberculosis using Affymetrix oligonucleotide GeneChips Journal of Infection.2007;54(S3):277-284 Fu LM, Shinnick TM. Genome-wide analysis of intergenic regions of mycobacterium tuberculosis H37Rv using affymetrix genechips. EURASIP journal on bioinformatics & systems biology.2007:23054

Saliva

Lucy E. DesJardin Isolation of M. tuberculosis RNA from Sputum Methods in Molecular Medicine.2001;48:133-139 Beenhouwer DO, Shapiro S, Feldmesser M et al. Both Th1 and Th2 Cytokines Affect the Ability of Monoclonal Antibodies To Protect Mice against Cryptococcus neoformans Infection and immunity.2001;69: 6445-6455 Desjardin LE, Perkins MD, Wolski K et al. Measurement of Sputum Mycobacterium tuberculosis Messenger RNA as a Surrogate for Response to Chemotherapy American journal of respiratory and critical care medicine.1999;160(1):203-10

Since the COVID-19 virus can be detected in stool samples, it has been proposed that detection of the virus in wastewaters will help to monitor local or regional outbreaks. For this to succeed, simple and robust measurement of viruses from wastewater is necessary. Conventionally, enteric viruses are first concentrated by a variety of filter methods. A secondary enrichment step is often employed, to reduce to a final volume and improve purity prior to final analysis. This secondary step may be provided by the Viraffinity™ reagent, and does not require ultracentrifugation. As described in these two references: 1) Viraffinity™ can dramatically improve the purification of viruses, and 2) can remove RT-PCR inhibitors in the detection of enteric viruses.

1) Romain Fragnoud R., et al. "Differential proteomic analysis of virus enriched fractions obtained from plasma pools of patients with dengue fever or severe dengue". BMC Infectious Diseases (2015) 15:518. http://www.ncbi.nlm.nih.gov/pubmed/26572220

The article's authors report a method to compare the proteomes of virion-enriched fractions purified from plasma pools of patients with dengue fever or severe dengue. Virions were purified by ultracentrifugation combined with Viraffinity™. “…and became more intense after the Viraffinity™ step...Densitometry indicated that the protein complexity of the purified samples was reduced by roughly 350-fold compared to the unpurified samples.”.

2) Leggitt, Paris R., and Lee-ann Jaykus. "Detection methods for human enteric viruses in representative foods." Journal of Food Protection 63.12 (2000): 1738-1744.

“To optimize viral nucleic acid amplification, secondary PEG precipitates… required a prior adsorption step with an equal volume of Viraffinity™ to further remove RT-PCR inhibitors. In this case, viruses …were adsorbed by the addition of an equal volume of Viraffinity™, … These Viraffinity™ precipitates were used directly in subsequent RNA extractions.”

Viraffinity™

Prep Size: Application dependent

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ViraPrep™ Mammal

Prep Size: 40 ml

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A research article describes the simplicity and efficiency of this virus enrichment technology to enhance viral proteomic analyses from plasma.

Romain Fragnoud R., et al. "Differential proteomic analysis of virus enriched fractions obtained from plasma pools of patients with dengue fever or severe dengue". BMC Infectious Diseases (2015) 15:518. http://www.ncbi.nlm.nih.gov/pubmed/26572220

The article's authors report a method to compare the proteomes of virion-enriched fractions purified from plasma pools of patients with dengue fever or severe dengue. Virions were purified by ultracentrifugation combined with Viraffinity™. “…and became more intense after the Viraffinity™ step...Densitometry indicated that the protein complexity of the purified samples was reduced by roughly 350-fold compared to the unpurified samples.". The authors conclude that the Viraffinity™ based technique of virion-enrichment allowed them to identify two host proteins that have prognostic value for classifying patients with acute dengue who are more likely to develop a severe dengue.