Protein Purification - Research Associate

Dmitry holds a B.S. and M.S., and he completed the Biotechnology Lab Specialist program at Shoreline Community College. He now works in protein purification at the Center for Infectious Disease Research in the South Lake Union area of Seattle, WA. Dmitry describes how he uses a variety of skills for conducting cutting-edge work to isolate and characterize proteins.

What biotechnology program did you attend?
Tambov State University, Tambov, Russia; Shoreline Community College

What degree/certificate(s) do you hold? 
B.S., M.S., Biotechnology Certificate from Shoreline Community College

What do you do for your job?
I oversee and coordinate a range of "high value target" projects, which involves cloning, expression, purification, crystallization, X-ray data collection and data analysis for solving 3D structure of 12 proteins from M. tuberculosis, L. donovani, Burkholderia cenocepacia.  I conduct assay development and optimization, protein expression, purification and analysis, and verification of experimental data. I also conduct maintenance and troubleshooting of lab equipment and problematic protein purifications. I also consult collaborators and other labs around the institution about protein expression and purification.

What are some techniques that you commonly use?

  • Assay development and optimization;
  • Experimental design;
  • Large scale protein expression;
  • Protein exclusion chromatography based on affinity, size, and ion-exchange; FPLC; HPLC; LC/MS;
  • Research of scientific literature;
  • Electronic data capturing systems and databases;
  • Biostatistics; DNA extractions;
  • Purification and analysis;
  • Molecular cloning;
  • PCR;
  • Western Blot;
  • IP;
  • ELISA;
  • Tissue culture.

Please describe what you do in an average day
Because I work to support both pipeline and special projects in our lab, I am involved with two main types of activities: doing standard protocols for protein purification pipelines and organizing research for so-called special projects, which are usually focused on a single protein or complex. For routine activity I maintain our FPLC machines, conduct lysis of bacterial pellets, use Immobilized Metal Affinity Chromatography and Size Exclusion Chromatography, analyze collected fractions via SDS PAGE gels and Western Blot, and enter data into a protein purification database. For special projects I will design particular experiments, communicate with PIs, collaborators, and subcontractors about project direction and progress, write reports and/ or papers, and make presentations for group meetings and conferences. 
 The schedule for a typical day is to:
-Lysis bacterial pellets, start new expression experiments
-Conduct FPLC prep and/or run, troubleshooting if nesessary
-Collect fractions, conduct SDS PAGE/Western
-Work with databases
-Send emails and complete paperwork
-Prep machines for next day/weekend storage

What advice would you give someone who is interested in a biotech career?
Determine the overall goal for your entire career, as well as short-term objectives (1, 5, 10 years increments)
-Determine your main scientific interest
-Find a mentor at each step of your career
-Try to always learn something new at each place you work (it's a hard part especially in the industry environment)
-Develop general scientific skills (critical thinking, paper writing etc.)
-Try to be an expert in at least one technical field.
-Be opportunistic
-If you are relatively young, consider grad school (but it's long commitment, so assess your risks)

Is there anything you'd like to add?
Everything stated above is based on my personal experience, so it may not work for everybody. However, I am happy if you found it helpful.

  • Adding lysis buffer into a beaker to break apart bacteria
  • In SDS PAGE/Western area prepping for gel loading
  • Loading protein samples at the beginning of a run.
  • Vortexing bacterial pellets for better homogenization
  • I'm placing thawed bacterial pellets into beakers for lysis
  • Deli case-prep work needs to be done before purifier run. That includes changing buffer containers, purging the lines, placing columns, setting the fraction collector.