Red Blood Cells
Dried blood spots for detection of autologous blood doping with targeted LC-MS/MS
Matt Kuruc1; Swapan Roy1; Johan Jakobsson2; Raik Wagner2; Christer Malm2; 1Biotech Support Group (BSG) LLC, Monmouth Junction, NJ; 2Pro Test Diagnostics AB (PTD), Umeå, Sweden
Dried Blood Spots Citations:
Michelle R. Robinson, Lei Guo; Raymond J. Gonzalez; Kara M. Pearson; Kevin P. Bateman; Daniel S. Spellman. Differentiating Modes of Drug Induced Liver Injury Using Parallel Reaction Monitoring LC-MS. ASMS Conference 2017 poster report. “Hemoglobin depletion improves PRM panel coverage in DBS and volumetric adsorptive microsampling (VAMS)”.
Hakuna, Lovemore, et al. "A simple assay for glutathione in whole blood." Analyst (2015). (http://pubs.rsc.org/en/content/articlelanding/2015/an/c5an00345h) “…Hb can be removed using a commercial product, HemogloBind™, which can isolate and remove up to 90% of blood Hb."
HemogloBind ™ Reference Applications –
Nguyen, Anthony T., et al.
"UBE2O remodels the proteome during terminal erythroid
differentiation." Science 357.6350
During reticulocyte maturation, the proteome is remodeled
through the programmed elimination of most generic constituents of the cell, in
parallel with abundant synthesis of hemoglobin. The study used multiplexed
quantitative proteomics to identify candidate substrates of UBE2O, an E2
(ubiquitin-conjugating) enzyme, in an unbiased and global manner. Because
of the overly abundant presence of Hemoglobin, selective depletion of
Hemoglobin was necessary. The article states “Reticulocytes were lysed
by vortexing for 5 minutes at room temperature… An additional 10 bed vol of
Hemoglobind™ suspension was added to the samples, which were then vortexed for
another 10 min at room temperature followed by 4 min of centrifugation at 10000
x g. The supernatants, which contain hemoglobin-depleted sample,
were … processed for TMT quantification.”.
Serum Exosome Analyses
Nishida-Aoki, Nao, et al.
"Disruption of Circulating Extracellular Vesicles as a Novel Therapeutic
Strategy against Cancer Metastasis." Molecular Therapy 25.1 (2017):
This study considers that therapeutic strategies
targeting cancer-derived extracellular vesicles (EVs) hold great promise
because of the possibility they reposition microenvironments to accommodate
metastasis. The researchers report on a novel strategy of therapeutic antibody
treatment to target cancer-derived EVs and inhibit the metastasis of breast
cancer in a mouse model. The article states “Hemoglobin was accumulated with
HemogloBind™ beads (Biotech support group, Monmouth Junction NJ, USA) followed
by 0.22 μm filtration. Then, the EVs in the sera were concentrated by
Complexes and Functional
Wan, B Borgeson, S Phanse, F Tu, K Drew, G Clark, et al. Panorama
of ancient metazoan macromolecular complexes. Nature Volume:525,
Pages:339–344 Date published:(17 September 2015). doi:10.1038/nature14877
Lahut, Suna, et al. "Blood RNA
biomarkers in prodromal PARK4 and REM sleep behavior disorder show role of
complexin-1 loss for risk of Parkinson's disease." Disease Models &
Mechanisms (2017): dmm-028035. http://dmm.biologists.org/lookup/doi/10.1242/dmm.028035
In this study, Parkinson’s disease
progression is investigated through the accumulation and aggregation of the
lipid-binding SNARE complex component alpha-synuclein (SNCA) which underlies
vulnerability and defines its stages. The authors studied blood samples from a
new large pedigree with SNCA gene duplication (PARK4 mutation), to
identify effects of SNCA gain-of-function as potential disease biomarkers. The
article states “For protein extraction from the EDTA tubes, 300 μl blood were
lysed with equal amount of 1% SDS-RIPA buffer [50 mM Tris-HCl (pH 8.0), 150 mM
NaCl, 1 mM EDTA, 1 mM EGTA, 1% Igepal CA-630 (Sigma), 0.5% sodium deoxycholate,
0.1% SDS, 1 mM PMSF and one tablet Complete Protease Inhibitor Cocktail
(Roche)] and sonicated for 10 sec. The blood lysates were rotated at 4 °C for
30 min and centrifuged at 4 °C for 30 min. The supernatants were depleted in
hemoglobin content using a commercial kit (HemogloBind™) following the
Katarína, et al. "Transketolase Activity but not Thiamine Membrane
Transport Change in Response to Hyperglycaemia and Kidney
Dysfunction." Experimental and Clinical Endocrinology &
Diabetes (2017). https://www.thieme-connect.com/products/ejournals/abstract/10.1055/s-0043-115009
Diabetic kidney disease, a common complication of both type 1 and type 2
diabetes, is associated with significant morbidity and mortality, and
represents the most common cause of chronic kidney disease. The study
hypothesized that protective pentose phosphate pathway action in diabetes might
be compromised by limited intracellular availability of an active transketolase
cofactor thiamine diphosphate (TDP). To evaluate the levels of thiamine
tranporter proteins in whole blood, t he article states “ For protein isolation, whole blood aliquots were lysed
with water and haemoglobin was removed using HemogloBind™ (Biotech Support
Group) according to manufacturer’s instructions…”.
Agnostic – Applications to Different Species
Snider, Thomas H., Christina M. Wilhelm, Michael C. Babin,
Gennady E. Platoff Jr, and David T. Yeung. "Assessing
the therapeutic efficacy of oxime therapies against percutaneous
organophosphorus pesticide and nerve agent challenges in the Hartley guinea pig."The
Journal of Toxicological Sciences 40, no. 6 (2015): 759-775.
Acetylcholine is an essential neurotransmitter, and inhibitors of
cholinesterases(ChEs) are potent toxins. A primary component of
anti-organophosphorus therapy is an oxime reactivator to rescue inhibited
acetylcholinesterases. For this, clinical signs of toxicity can be measured
from blood cholinesterase [Acetylcholinesterase (AChE) and
butyrylcholinesterase (BChE)] activity utilizing a modified Ellman's method.
Biotech Support Group’s unique solid-phase polymer for hemoglobin depletion,
was used for pretreatment. The article states “Briefly, whole blood samples
were treated with HemogloBind™ which interferes with the ChE activity assay due
to spectral overlap.”
Craig, J. R., et al. "A comparison of the anatomical and gastrointestinal
functional development between gilt and sow progeny around birth and weaning." Journal
of animal science (2019).
Gilt progeny (GP) often have
restricted growth performance and health status in comparison to sow progeny
(SP) from birth. To better understand underlying mechanisms, the study aimed to
compare differences in growth and development between GP and SP in the first 24
h after birth and in the peri-weaning period. Because serum
samples were quite hemolysed after collection and processing, it became
necessary to use HemogloBind™ to allow for better detection of IgG by ELISA. The
article states “As per the manufacturer’s instructions, 250 μL of Hemoglobind
was added to 250 μL of hemolyzed serum…
Parvathi S. Kumar, Haree K. Pallera, Pamela S. Hair, Magdielis
Gregory Rivera, Tushar A. Shah, Alice L. Werner,Frank A. Lattanzio, Kenji M.
Cunnion, and Neel K. Krishna. Peptide
inhibitor of complement C1 modulates acute intravascular hemolysis of
mismatched red blood cells in rats.TRANSFUSION Volume 00,
May 2016. doi:10.1111/trf.13674.
In brief, the study evaluated the role of a peptide inhibitor of complement C1
(PIC1) in an animal model of acute intravascular hemolysis in both prevention
and rescue scenarios. The authors state "To remove free Hb that may cause
optical interference in bilirubin analysis, we treated all the samples with Hb
depletion from hemolyzed serum/plasma (HemogloBind, Biotech Support Group).
Bilirubin concentration was then measured with a Bilirubin Assay Kit
(Sigma-Aldrich, St. Louis, MO)."
Laing, Richard W., et al.
"The use of an acellular oxygen carrier in a human liver
model of normothermic machine perfusion." Transplantation 101.11
Red cell lysates
O’Connell, Grant C., et al. "Monocyte-lymphocyte cross-communication via soluble CD163
directly links innate immune system activation and adaptive immune system suppression
following ischemic stroke." Scientific
reports 7.1 (2017): 12940
Kyoungsook Park, Christopher D. Saudek, and Gerald W.
Expression of β-N-Acetylglucosamindase (O-GlcNAcase) in Erythrocytes from
Prediabetic and Diabetic Individuals.Diabetes.2010;59(7):1845-50.
Delobel J., Rubin O., Prudent M., Crettaz D., Tissot J.-D., Lion N.(2010) Biomarker Analysis of Stored Blood
Products: Emphasis on Pre-Analytical Issues. International
Journal of Molecular Sciences. 11(11):4601-4617
Alvarez-Llamas, Gloria, Fernando de la Cuesta, Maria G.
Barderas, Irene Zubiri, Maria Posada-Ayala, and Fernando Vivanco. "Characterization
of Membrane and Cytosolic Proteins of Erythrocytes."
In Vascular Proteomics, pp. 71-80. Humana Press, 2013.
Alvarez-Llamas, G., de la Cuesta, F., Barderas, M. G., Darde, V. M., Zubiri,
I., Caramelo, C., Vivanco, F. A novel
methodology for the analysis of membrane and cytosolic sub-proteomes of
erythrocytes by 2-DE.Electrophoresis.2009;30:4095-4108
Zihao Wang, Kyoungsook Park, Frank Comer1, Linda C.
Hsieh-Wilson, Christopher D. Saudek, Gerald W. Hart. Site-Specific GlcNAcylation of Human
Erythrocyte Proteins: Potential Biomarker(s) for Diabetes Mellitus.
Yuichi Miki, Tomoki Tazawa, Kazuya Hirano, Hideki Matsushima,
Shoko Kumamoto, Naotaka Hamasaki, Tomohiro Yamaguchi, Masatoshi Beppu. Clearance of
oxidized erythrocytes by macrophages: Involvement of caspases in the generation
of clearance signal at band 3 glycoprotein.
Biochemical and Biophysical Research Communications.2007; 363(1):57-62
quantification of glycated Cu-Zn superoxide dismutase in erythrocytes by
electrospray ionization mass spectrometry,
Biochimica et Biophysica Acta. 1999.1426(3):483-90