Unraveling Neurodegenerative Diseases with Jess

A DIVERSE AND ACCOMPLISHED CAREER

Over the course of Dr. Nicolas Bazan’s several decades-long career, he has been a research scientist, teacher, mentor, community leader, author, screenplay co-writer, executive movie producer, patron of the arts, and entrepreneur. Currently, he is serving as Director of the Neuroscience School of Excellence at the leading Louisiana State University School of Medicine. Here, his research seeks to unravel the cellular and molecular events in neuroinflammatory lipid signaling in early stages of neurodegenerative diseases like Alzheimer’s disease, dementia, and other brain and retinal dysfunctions. He also studies how elovanoids and related molecules can serve as potential therapeutics to prevent deleterious effects of SARS-CoV-2 infection in human lung tissue, nasal mucosa, the eye, and the brain.

Dr. Bazan, who has published close to 500 peer-reviewed publications, sings the praises of Simple Western assays with Jess. Why? As a long-time user of traditional Western blot, he was all too familiar with Western blot’s notorious limitations and challenges. Instead, Jess has dramatically improved the protein analytical methods on which his laboratory relies to study the cellular and molecular events of neurodegenerative disease onset and progression. With Jess, he has successfully quantified proteins and peptides from a variety of tissues, cultures and other types of samples including IL-1 beta, SARS-CoV-2 spike protein, ACE2, sirtuin, furin and iduna.

NO TIME FOR WESTERN BLOTS

If you’re busy studying neurodegenerative diseases like Dr. Bazan, you don’t have time for traditional Western blots. Every advantage is needed when studying Alzheimer’s disease and the neuroscience of aging. The Jess instrument allows neuroscience researchers to automate these traditional blots and focus their time on studying the cellular and molecular mechanisms related to neurodegenerative diseases.

With traditional Western blot, Dr. Bazan describes, “it took us two days plus to obtain results and the maximum samples run per person were 60.” Frustrated with low throughput and slow time to results, Dr. Bazan’s lab turned to Jess to automate their workflows. They were so impressed that they now regularly run not just one, but three Jess instruments fulltime. “Using three Jess systems running twice a day we can run 250 samples in two days.” Thanks to the high throughput of Jess, they can “breeze through screening samples and validating targets.” He exclaimed, “it saves so much time! It takes away so much of the pipetting and manipulation error. You just load your samples and wait for results, and in the meantime, you can dedicate yourself to other activities.” Also, the built-in analysis software, Compass for Simple Western, is “a very good tool to navigate and compare different samples, without having to pass from one equipment to another, for instance gel doc and computer to analyze results.”

GETTING THE MOST DATA PER SAMPLE

With traditional Western blot, Dr. Bazan was having problems with stripping and re-probing. “Stripping and re-probing also brought the issue of signal loss.” Instead, Dr. Bazan and his colleagues rely on Jess’s newest feature, RePlex, which efficiently removes antibodies from the first round of probing for a second round of immunoassay, all in an automated fashion. Because the proteins are covalently bound to the capillary wall, there is no signal loss between probing cycles.

The second probing cycle may also be dedicated to Total Protein Detection, which is critical for normalizing protein expression data, especially between blots. As Dr. Bazan explains, “it was also a challenge to compare sets of 15 or more samples, because they were run in different blots and the comparisons usually were not accurate.” For this, “total protein standardization allows us to compare different runs seamlessly.”

SAVING MOUSE TEARS AND OTHER PRECIOUS SAMPLES

Dr. Bazan noted that perhaps the “most important feature of all” is that Simple Western requires very small sample volumes – as little as 3 μL. With traditional Western blot they would often have to pool different individuals together to get enough sample, which interfered with statistical analysis. Fortunately, Jess was designed for small sample volumes, allowing them to detect proteins in mouse tears and other precious samples without the need to pool individuals together. “We are able to get protein expression/abundance data from several experimental models including cell cultures (human alveoli in culture) and animal studies that would be difficult with traditional WB because of the size of the samples.” Importantly, scaling down sample size did not interfere with the quality of the data. “The reproducibility and sensitivity are good, in contrast to some of the methods used for traditional Western blots. The accuracy also let us distinguish variations that were not noticeable with traditional Western blots.”

STOPPING THE STIGMA OF BRAIN DISEASES

Dr. Bazan is an advocate of science and of removing the stigma of neurodegenerative diseases like Alzheimer’s disease and dementia. To fulfill these goals, he has written novels exploring his lifelong intellectual quest to understand the deep beauty and complexity of brain function and the human experience. These novels include The Dark Madonna: A Fable of Resiliency and Imagination and Una Vida: A Fable of Music and the Mind. The latter novel was adapted into a feature film for which he was the screenplay co-writer and Executive Producer, called Of Mind and Music.

Dr. Bazan has implemented translational approaches for his discoveries on signals that allow understanding the rules of the life of brain and retina cells. He and his colleagues have uncovered that these signals made up inflammatory responses that evolve during aging and after injury. He began to dissect these molecular responses in stroke, TBI, pain, and other neurological and ophthalmological conditions, particularly in the early stages of Alzheimer’s disease and of age-related macular degeneration. The rules of life he postulated are driven by low abundance but very high potency mediators that protect the integrity and survival of the brain and retina at the onset of diseases. Jess is a helpful tool to decipher these signals. Applying these discoveries, he is the founder of 3 new startup companies (neuresto.com, southrampartpharma.com, and curvirbiotech.com).

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