Authors list:
Binesh Shresta

Abstract:

Baculovirus mediated insect cell expression system is one of the most popular eukaryotic expression systems for protein production. This has been a main workhorse to fulfill high demand of recombinant protein needed for structural, biophysics and biochemical studies. The eukaryotic system has been in use for more than 2 decades. Many of the methods developed at inception are still routinely used. Different labs have built up in-house experience over the time to customize their work flow with incremental modifications. This might be the reason the protocol developed in one lab is not easily reproducible in another when it comes to deal with targets that are difficult to express or protein complexes which requires attentive details. Here we tried to share the experience of our lab from last 14-15 years that may be useful for new user of insect cell expression system. First and foremost the important part we need to consider is how the cells are grown and maintained over the time. We have observed cells maintained in different medium over long period of time has a different properties. It is noteworthy to remember if cells were maintained with a long passage number you may see the superior quality that might be useful for you. This might have happened by clonal selection over the time without our notice.