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Improving Trauma Outcomes with Fast, Efficient Brain Injury Biomarker Analysis

""We were fortunate to be able to test these Luminex porcine panels early in our research effort. Once we could see the measurements we were getting on the plates, we knew we had found what we were looking for. The panels ended up making a huge difference in the outcome of the study." "

- Elizabeth Rick, Research Scientist, Oregon Health & Sciences University (OHSU)

LUMINEX® ASSAYS HELP ADVANCE BRAIN INJURY BIOMARKER PROFILING RESEARCH

Traumatic brain injury (TBI) is among the leading causes of death and disability in the United States, with an estimated 2.8 million individuals seeking medical treatment of TBI each year.1 Available treatment options for TBI are limited, with current strategies focused primarily on reactive management of initial symptoms and secondary effects.

Elizabeth Rick is a research scientist at Oregon Health & Sciences University (OHSU) working in the division of Trauma, Critical Care and Acute Care within the Surgery department. The OHSU Trauma Division includes an active clinical and basic research program focused on bringing innovation to the treatment and care of trauma patients, not just at OHSU but around the world.

Recent studies, including a large, clinical, randomized study of TXA administration in head trauma directed by the Trauma lab at OHSU, have shown reduced mortality and improved neurological outcomes with early administration of tranexamic acid (TXA)—a potent antifibrinolytic that inhibits enzymatic clot breakdown.2 Gaining a deeper understanding of the relationship between TXA and endothelial disruption in patients with TBI is a core focus for the research team.

IMPROVING DISCOVERY OF TBI BIOMARKERS WITH BETTER ASSAY SENSITIVITY

While decreased mortality has been attributed to reduction in intracranial hemorrhage (ICH), there’s growing interest in characterizing the possible mechanisms associated with this reduction and the effect of TXA in helping to prevent secondary injury.3  In a recent study examining the effect of early TXA administration on circulating markers of endotheliopathy, the research team at OHSU was able to catalog many of the endothelial markers and cytokine concentrations in patients with moderate-to-severe traumatic brain injury (MS-TBI) and ICH.

The need to quantify biomarkers quickly and efficiently and correlate them to specific outcomes is a critical requirement across all areas of translational research. In this case, the OHSU team found the optimum solution with Luminex Discovery assays from Bio-Techne.

The cost-efficient screening capabilities of the multiplex format provided researchers with a fast and effective way to simultaneously profile endothelial and coagulation activation—two key factors in the preservation of endothelial integrity. Meanwhile, the flexible Luminex configuration options allowed researchers to choose from a large menu of analytes to build their custom assay panel.

“The superior accuracy and sensitivity of the Luminex panels proved instrumental in giving us the high-quality data we needed,” Rick said. “Once we identified our targets and could see the concentrations we were getting, this increased our confidence in the results indicating improved endothelial function.”  

 

FINDING THE RIGHT PORCINE ASSAY

The Trauma Research group additionally uses a pig model to improve insight into endotheliopathy and the endothelial damage following trauma.  This focus demands robust assays, particularly in the animal model but suitable porcine panels have been difficult to find largely because the porcine model is not as frequently used as the rodent models. 

“The porcine model is not a universal model so there are not a lot of multiplexing options,” Rick said. “We’ve worked with several vendors to try and set up a porcine cytokine assay in a multiplex format, but all of them have come up short. We would run the assay and there would be no consistent, robust concentrations on the plate.” 

In one recent study analyzing the effect of stem cell treatment to prevent or lessen the possibility of acute respiratory distress syndrome (ARDS) following traumatic hemorrhage and injury, the Trauma Research team found themselves once again searching for a suitable porcine panel in a multiplex assay format that could meet their detection requirements.  They ultimately found what they were looking for, thanks to its close partnership with Bio-Techne.

While talking with her Bio-Techne sales rep in 2019, the rep happened to mention some new Luminex porcine panels that were currently in development. Thinking quickly, Rick asked the rep if it was possible to get early access to some of the plates that were still in beta testing.

“We were fortunate to be able to test these Porcine Luminex Discovery Assays early in our research effort,” Rick said. “Once we could see the measurements we were getting on the plates, we knew we had found what we were looking for. The panels ended up making a huge difference in the outcome of the study.”  

 

BUILDING EARLY CONFIDENCE IN THEIR RESEARCH RESULTS

As part of her comprehensive research analyzing collected samples for biomarkers of brain injury, endotheliopathy, and traumatic hemorrhage effects, Rick has experimented with a variety of different assays and platforms. In one recent project, Rick and her team employed a combination of Luminex assays and Quantikine™ ELISAs from Bio-Techne.

For this particular project, Rick was tasked with training a group of interns from the local university on how to set up and run several of the ELISAs. Even though the setup and operation of the assays were all new to them, it didn't matter, Rick said, because of the easy-to-follow protocols and the exceptional accuracy of the assays.

“We set up and ran 10 plates,” Rick said. “The curves were beautiful, and the results were unbelievably accurate every time. It was really fun to work with this group —they now think that all ELISAs run that flawlessly.”  

 

GAINING AN ACCURACY AND CONSISTENCY ADVANTAGE

The ease of use and transparency of both the Luminex and ELISA assays has proven to be an important advantage for the research team.

“From the first time we looked at the Bio-Techne plates, we noticed a clear difference in the clarity of the protocols and procedures,” Rick said. “When it comes to overall assay consistency and accuracy, there’s no comparison,” Rick said. “We keep coming back to Bio-Techne because we can’t find that quality anywhere else.”

Transitioning from plate-based ELISAs to the Porcine Luminex Discovery assays allowed Rick and her team to take advantage of the efficiency and throughput advantages of the multiplex format.

“It’s not just the cost of the plates, but also the time required to set up a larger number of ELISAs,” Rick said. “We don’t have a large in-house staff, so when we found the Luminex porcine panels, it was really exciting.”

Assessing multiple analytes in a single assay also allows Rick and her team to be judicious with materials while generating much more useful information, alleviating the need to prioritize assays based on the volume of precious samples and allowing for future studies using those same samples.

With access to the right analytical tools, Rick can continue doing translational research that impacts lives while remaining confident in her ability to validate results across different types of studies and between different assay platforms. In the future, the research team plans to use the Luminex Discovery assays with human models to perform additional analysis on endothelial markers and cytokines in TBI and trauma patients.

REFERENCES

  1. Taylor CA, Bell JM, Breiding MJ, Xu L. (2007 and 2013) Traumatic brain injury-related emergency department visits, hospitalizations, and deaths—United States, MMWR Surveill Summ. 66(9):1–16.
  2. Zehtabchi S, Abdel Baki SG, Falzon L, Nishijima DK. (2014) Tranexamic acid for traumatic brain injury: a systematic review and meta-analysis. Am J Emerg Med. 32(12):1503–1509.
  3. Jokar A, Ahmadi K, Salehi T, Sharif-Alhoseini M, Rahimi-Movaghar V. (2017) The effect of tranexamic acid in traumatic brain injury: a randomized controlled trial. Chin J Traumatol. 20(1):49–51.
  4. Akwii RG, Sajib MS, Zahra FT, Mikelis CM. (2019) Role of angiopoietin-2 in vascular physiology and pathophysiology. Cells. 8(5):471.

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