Protein production and purification.
Structural Genomics Consortium None., China Structural Genomics Consortium None., Northeast Structural Genomics Consortium None., Gräslund S., Nordlund P., Weigelt J., Hallberg BM., Bray J., Gileadi O., Knapp S., Oppermann U., Arrowsmith C., Hui R., Ming J., dhe-Paganon S., Park H-W., Savchenko A., Yee A., Edwards A., Vincentelli R., Cambillau C., Kim R., Kim S-H., Rao Z., Shi Y., Terwilliger TC., Kim C-Y., Hung L-W., Waldo GS., Peleg Y., Albeck S., Unger T., Dym O., Prilusky J., Sussman JL., Stevens RC., Lesley SA., Wilson IA., Joachimiak A., Collart F., Dementieva I., Donnelly MI., Eschenfeldt WH., Kim Y., Stols L., Wu R., Zhou M., Burley SK., Emtage JS., Sauder JM., Thompson D., Bain K., Luz J., Gheyi T., Zhang F., Atwell S., Almo SC., Bonanno JB., Fiser A., Swaminathan S., Studier FW., Chance MR., Sali A., Acton TB., Xiao R., Zhao L., Ma LC., Hunt JF., Tong L., Cunningham K., Inouye M., Anderson S., Janjua H., Shastry R., Ho CK., Wang D., Wang H., Jiang M., Montelione GT., Stuart DI., Owens RJ., Daenke S., Schütz A., Heinemann U., Yokoyama S., Büssow K., Gunsalus KC.
In selecting a method to produce a recombinant protein, a researcher is faced with a bewildering array of choices as to where to start. To facilitate decision-making, we describe a consensus 'what to try first' strategy based on our collective analysis of the expression and purification of over 10,000 different proteins. This review presents methods that could be applied at the outset of any project, a prioritized list of alternate strategies and a list of pitfalls that trip many new investigators.