The Great Flask-tastrophe

As a master’s student at Colorado State University in the late 1980s, I worked on developing a purification protocol for an antiviral enzyme, 2-5A synthetase.¹ This enzyme, induced by interferons, targeted viral RNA with an RNase, and was enriched in immature red blood cells called reticulocytes. 

Collecting large amounts of rabbit reticulocytes took weeks of preparation. On one occasion, I isolated the reticulocytes and suspended the cells in a low-salt lysis buffer in a large glass flask like normal. But then, disaster struck when I set the flask a bit too hard on the benchtop.  

I watched in horror as my red blood cell lysate bled on the white absorbent paper bench coat. Desperate not to lose the last two months of work, I quickly realized that the paper was primarily cellulose, and the next step of my protocol involved running the sample through a cellulose column. I carefully curled up the ends and wrung the lysate into another flask. Although I lost a quarter of my total volume, I figured finishing the rest of the protocol with my recovered sample was worth a shot.

I never told my boss about this harrowing ordeal, but as it turned out that lysate was the best prep of 2-5A synthetase during my graduate program. It had high purity, low protein, and high enzymatic activity—exactly what we were after. 

Looking back, it turned out not to be an epic failure, but it sure felt like one in the moment. Many students get demoralized by setbacks, but in science, things go wrong all the time. As I concluded my career with a greater emphasis on teaching, I realized how important it was to communicate these concepts like perseverance to my students. Even in retirement, the lesson to never give up remains with me from my career. 

Joel Rovnak is now retired and an emeritus professor at Colorado State University. This interview has been edited for length and clarity.

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