Azari Guerrero, Burton Lab at Scripps Research, Week 6

Welcome back! I’m Azari, and I just completed my final week at Scripps Research. Just a refresher, I worked with Nate Beutler, a post-doctoral immunologist who, through antibody discovery and therapeutic design, is working to understand why an effective vaccine for malaria hasn’t been developed yet. I also worked with Ben Nemoz, a post-doc virologist who specializes in HIV research. He holds a PhD in the United States and an MD in France, so he splits his time between both places. This week I wrapped up my final project, presented to the lab, and said my final goodbyes to everyone and everything that made this experience so amazing.

Monday morning we helped prep an ELISA for Yenni while he ran some errands. All of our mentors had something to work on so we sat in Nate’s office all day and did a bunch of research. I read four long research papers about all the complexities of malaria, specifically the parasite Plasmodium falciparum. They were kind of hard to follow but I was able to understand the parasite’s lifecycle by the end of the day. I also worked on my presentation I would be giving to the lab. During my first week I briefly mentioned to Nate that each Pintern has to do a presentation later on, so Nate had me write about what I did and practice presenting in front of the lab.

One of the neutralization plates

Shiloh and I started Tuesday with Ben as he began the first steps of the neutralization assay, our last project in the lab. The point of a neutralization assay is to test if an antibody is able to neutralize (block) a virus from binding to a host cell. If it can’t the cell will glow, if it can it won’t glow. The cells are engineered to glow when a certain enzyme in the virus enters. The first steps are to combine the virus, antibody, and cell into a media that is added to 20 well plates. We did a serial dilution for each of the plates too to see at which concentration the antibody is most effective. This has to be incubated for 48 hours so our work with Ben ended there for the day. Nate and Sean took us to Raising Cane’s because it was Shiloh’s last day in the lab (thanks guys!). Back at the lab, I presented my slideshow of all the work I did during the past 6 weeks. I was very nervous but everyone there was so nice which made it much easier, I’ll definitely miss how kind the lab members are. We still had a bit of time left so we helped Nate build a contraption to hold a hamster he would be testing on. It turned out great so we went downstairs to get our hamster and tested on him. We named him Albert. Nate was testing to see if Trypan Blue could get into it’s lungs; if successful he’d repeat the experiment on five more hamsters but with SARS-Co-V2, the virus responsible for Covid-19, instead. We humanly euthanized our hamster to be dissected. By the way, listen to people when they say you shouldn’t name your hamsters because we did end up getting attached. We sang to Albert as he passed and I might have shed a tear or two. We finished up and it was time for Shiloh to return her lab coat and say goodbye to the lab. We’ve both grown to see the lab like a second home so it was very emotional watching her leave for the last time.  That night we had our last meal all together and got ice cream. Shiloh and I also had our last nightly walk, a sort of habit we picked up, and encountered a free library where we each took a book from. Shiloh had to leave at 4 am so we all woke up to say bye. It was definitely sad to see her go but we’re planning on reuniting later this summer!

UCSD campus!

There wasn’t much work for me on Wednesday since our neutralization wasn’t ready yet and Nate had animal work to do at UCSD. While I wasn’t allowed to go in to watch the experiment, Nate still took me with him so I could explore the campus. I’ve been seriously considering going to college in San Diego so I’m grateful I was able to wander around and see everything in person. I spent the rest of the day eating lunch and working on personal stuff. I didn’t get to do much science but I’m happy I was still able to be productive. Luckily the neutralization was ready on Thursday so I spent all day doing wet work. First I vacuumed out the media, then I lysed the cells and added substrate together (lyse opens up the cell so we can see the glow, substrate is what excites/causes the glow). It’s very light sensitive so we have to be quick when running it through the luminescence plate reader. There are 20 plates so this process is very tedious and time consuming, but it was still super cool being involved in the experiment. Ben let me do it all on my own so it felt great seeing the consistent results. We found that one of the more mature antibodies was the most effective in neutralizing all of the viruses which Ben described as ‘puzzling’. We also had a bit of time to put all of our data on the computer. I’m not the best with the technological side of STEM but I think I got the hang of it eventually! The end goal is to get a percentage of how effectively each antibody neutralized the viruses. 

The results of one of our neutralization plates

Friday morning I finished analyzing the data with Ben which took about 2 hours. He put the results through Python to create an easy to visualize scatterplot since the other program he uses isn’t very good at that. Our scatterplots showed that antibody PC94-A49 was the most effective in neutralizing our viruses. Yenni was kind enough to bring me pasta for lunch as a little going away gift; it was very good. My last task was helping Nate prep his sample for flow cytometry. He wanted to analyze animal blood cells that were infected and treated with different drugs. Ben then gave me a lesson on blood types and how blood tests work. I think it’s super cool when he shares his medical knowledge with me too! Nate had to leave a bit early so I said my goodbyes to him then. I may or may not have cried a little saying bye. I ended the day by saying my goodbyes to Yenni and Ben which was also really emotional for me. I walked out and drove off for the last time secretly hoping someone from the lab would walk by so I’d have an excuse to go back and stay.

My last night in San Diego, I miss it already!!

This internship has been one of the best experiences of my life. Going in I was terrified about the thought of leaving home to pursue my education and now I’m leaving counting down the days until I can go back. I’ve not only learned so much about immunology, but I’ve also learned so much about myself. This experience has me seriously considering applying to UCSD and majoring in a similar field. I did not have a single bad day even when the work got difficult. I want to give a huge thanks to everyone involved in making this the best summer. Shout out Sarah, Trang, and Jennie for making this internship possible! And of course, thank you Nate, Ben, Yen-Chung, and everyone else in the lab for taking me in and sharing your love for science with me. It’s been so inspiring working with you all and anyone would be lucky to have the opportunity to work with you. You’ve all changed my life for the better. Finally, I want to give a huge thanks to Team SD, you’re all so amazing and talented and I cannot wait until we reunite. I got that sneak peak of my future I was hoping for and I cannot wait until I get to go off and live it.

Thanks for reading : )