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The massive amount of film that I used to go through was an inconvenience, which now has been completely eradicated by this little machine

Dr. Vikash Reebye
Imperial College London, UK

At the intersection of molecular biology and surgery, Dr. Vikash Reebye is formulating novel remedies to combat degenerative liver diseases and liver cancer. The Senior Research Fellow in Professor Nagy Habib's lab at the Imperial College London, UK, is translating bench-side research into budding therapeutic candidates for the bedside.

Kasinski Lab Uses Odyssey CLx and Infrared EMSAs to Study RNA

In an attempt to target loss-of-function diseases, the lab is developing small activating RNA (saRNA) to reintroduce protein expression; for instance, albumin, which declines in cirrhosis and hepatocellular carcinoma (HCC)1. “We inject HCC animal models with our treatment and look at preclinical parameters such as expression of albumin and liver enzymes1,” he says. Similarly, the group is also evaluating techniques to differentiate primary hematopoietic adult stem cells for transplantation in degenerative liver diseases.

Dr. Reebye assesses the protein expression levels in his preclinical samples with the snap of a ‘digit.’ The lab houses a C-DiGit® Blot Scanner, which is widely used for Western blotting.

An Innovative Technology for Analysis of Protein Expression

According to Dr. Reebye, the C-DiGit Blot Scanner is a great innovation, and it has changed their lab's workflow remarkably. They probe blood collected from animals [injected with the saRNA albumin treatment], for the presence of albumin using Enzyme-Linked Immunosorbent Assay (ELISA). “ELISA tells us if the liver function has regenerated to the point where it is secreting albumin into circulation. Our RNA is supposed to increase the target (albumin). Then, we look at target expression at the RNA level in a biopsy sample, and this is where the Western blot comes in.” Alpha-fetoprotein, enzyme ATT, and other biomarkers are similarly profiled using both ELISA and Western blotting. “Basically, the Western blot validates everything that we read from the blood [ELISA] as well as the transcript signatures that we get from the organ.”

Together with the Image Studio™ Software for the C-DiGit Blot Scanner, Dr. Reebye is able to perform a semi-quantitative protein expression analysis. “The software is very easy to use. It is sufficient for us to measure band intensity and say that the protein level reflects very nicely with the [RNA] transcript level and blood analysis obtained using ELISA.”

User-Friendly Instrument with a Small Footprint

I started just using the C-DiGit Scanner myself, but now everybody wants to use it.

The C-DiGit Blot Scanner has also allowed the lab to reclaim their darkroom space. “London is a very tight city, so real estate in the lab is valuable. The tiny C-DiGit Scanner has given me a lot of space, as I was able to scrap my darkroom and place a microscope there.”

Admitting his dislike of spending time in the darkroom, he says, “I was desperately trying to get out of it [darkroom]. Also the consumables; the massive amount of film that I used to go through was an inconvenience, which now has been completely eradicated by this little machine.”

1Vikash Reebye, Pål Sætrom, Paul J. Mintz, Kai-Wen Huang, Piotr Swiderski, Ling Peng, Cheng Liu, Xiaoxuan Liu, Steen Lindkær-Jensen, Dimitris Zacharoulis, Nikolaos Kostomitsopoulos, Noriyuki Kasahara, Joanna P. Nicholls, Long R. Jiao, Madhava Pai, Duncan R. Spalding, Malkhaz Mizandari, Tinatin Chikovani, Mohamed M. Emara, Abdelali Haoudi, Donald A. Tomalia, John J. Rossi and Nagy A. Habib. Novel RNA oligonucleotide improves liver function and inhibits liver carcinogenesis in vivo (2014). Hepatology Volume 59, Issue 1, pages 216–227, January 2014. DOI: 10.1002/hep.26669.

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