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Species Diversity Supported By BSG Products

Species Diversity Supported By BSG Products

With many products and alternative strategies for depletion and enrichment, Biotech Support Group offers distinct and diverse separations tools that work with virtually all species

Background

We are often asked “Can our products be used on different species?

The simple answer is yes. Biotech Support Group (BSG) is an innovator in the supply of proteomic sample preparation products. Our central technology focus is to develop highly selective separations products that do not rely on immuno-affinity. These can offer many advantages when working with human samples as well as other species. In fact, we have not come across a species whereby our products did not perform well for the applications required of it.

The Challenge

As a way to deplete high abundance proteins, many proteomic enrichment strategies employ the use of immuno-affinity depletion to remove one or more high abundance proteins. Some common limitations of immuno-affinity however are high costs, regeneration requirements which may result in a diminished and inconsistent performance, as well as a required marriage of species to antibody. Because of these limitations, researchers need ways to enrich differently.

The Solution

BSG has developed bead separations tools that do not use immuno-affinity and so can be adapted to virtually all species. While we have many references on human, in this case study, we highlight recent publications for all other species for:

  • Hemoglobin Removal
  • Albumin Removal
  • Lipid Removal

The Outcomes

Outcome #1

  • Sample Type: Hemolyzed pig serum
  • BSG Product: HemogloBind™
  • Analytical Platform: ELISA

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).

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, vortexed for 30 s before mixing via inversion for 10 min, and then centrifuged at 6,000 g for 2 min at 4°C. Purified serum was then aspirated and assessed in duplicate for IgG concentration … using a commercial ELISA kit.


Outcome #2

  • Sample Type: Mouse Serum
  • BSG Product: AlbuVoid™
  • Analytical Platform: LC-MS

Poillet-Perez, Laura, et al. " Autophagy maintains tumour growth through circulating arginine." Nature (2018): 1.

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.


Outcome #3

  • Sample Type: Frog plasma
  • BSG Product: AlbuSorb™
  • Analytical Platform: LC-MS

A U.S. EPA laboratory is building plasma protein ion libraries of the frog, Xenopus laevis, to quantitate protein-level changes between sexes and throughout development to serve as the foundation for evaluating phenotypic changes in X. laevis sex when exposed to various chemicals. The poster entitled

"High resolution accurate mass spectrometry-based proteomics in ecotoxicology: SWATH-MS to detect differentially expressed plasma proteins in the amphibian toxicological model Xenopus laevis"

states "Plasma samples from individuals of all ages: albumin-depleted (AlbuSorb™ Biotech Support Group), digested, reduced and alkylated, peptide-level cleanup (Oasis MCX SPE column, Waters), aliquoted at 0.1 µg/µl for online 1D gradient separation (2D Eksigent nanoLC with cHiPLCNanoflex C18 trap-elute columns @ 35°C)"


Outcome #4

  • Sample Type: Sheep serum
  • BSG Product: Cleanascite™

Analytical Platform: in vitro maturation (IVM) of ovine oocytes

Barrera N, dos Santos Neto PC, Cuadro F, Bosolasco D, Mulet AP, Crispo M, et al. (2018) Impact of delipidated estrous sheep serum supplementation on in vitro maturation, cryotolerance and endoplasmic reticulum stress gene expression of sheep oocytes. PLoS ONE 13(6): e0198742. https://doi.org/10.1371/journal.pone.0198742

The authors wanted to determine whether the use of delipidated estrous sheep serum during in vitro maturation (IVM) of ovine oocytes reduces the cytoplasmic lipid droplets content and improves embryo development and cryotolerance after vitrification. The article states “Lipid removal from serum was performed by using Cleanascite™ (Biotech Support Group, NJ, USA) according to the instructions provided by the manufacturer. Unlike other approaches, the protocol described herein for delipidation of estrous sheep serum was effective in decreasing levels of Triglycerides, total Cholesterol, and NEFAs. To our knowledge this is the first study to use the Cleanascite method to generate estrous sheep serum yielding significantly reduced lipid levels. Subsequent use of the partially delipidated serum as supplemented in IVM media resulted in effective reduction of oocyte lipid content. The advantage of this method over other traditional methods (i.e. chloroform) includes increased feasibility and reduced toxicity and biosafety concerns”.


Outcome #5

  • Sample Type: Fish Red Cells
  • BSG Product: HemoVoid™
  • Analytical Platform: LC-MS

Puente-Marin, Sara, et al. " In Silico Functional Networks Identified in Fish Nucleated Red Blood Cells by Means of Transcriptomic and Proteomic Profiling." Genes 9.4 (2018): 202.

Nucleated red blood cells (RBCs) of fish have, in the last decade, been implicated in several immune-related functions, such as antiviral response, phagocytosis or cytokine-mediated signaling. Label-free shotgun proteomic analyses were carried out for in silico functional pathway profiling of rainbow trout RBCs. The article states “The cytosolic fraction, approximately 300 μL, was depleted of hemoglobin using HemoVoid TM kit (Biotech Support Group, Monmouth Junction, NJ, USA), in accordance with the manufacturer’s instructions”.


For more information on the complete line of Albumin & IgG Removal products, visit: https://www.biotechsupportgroup.com/Articles.asp?ID=451

For information on all Hemoglobin Removal products: visit:

https://www.biotechsupportgroup.com/Articles.asp?ID=452

For more information on Lipid Removal Reagent and Clarification, visit:

http://www.biotechsupportgroup.com/Cleanascite-Lipid-Removal-Reagent-p/x2555.htm