Fig. 3: Crystallization capabilities of insect cell lines.

a Visual comparison of crystals in Sf9 and High Five cells, illustrating the size differences between crystals of the same protein produced in the two cell lines. Representative images of three independent experiments are shown. High Five cells produce significantly larger crystals than Sf9 cells while the morphology remains identical. Measurements of several hundred crystals in the right-side panels illustrate the large size differences. Yellow dots, Sf9 cells; violet dots, High Five cells. b High Five cells also show improved crystallization capabilities compared to Sf9 cells characterized by a higher fraction of crystal-containing cells within an infected culture, while no differences are visible regarding the fraction of infected, EYFP-producing cells. c An MOI of 0.5 is sufficient for infection of and protein production in nearly all cells within a High Five cell culture. However, a MOI of 1 should be used to maximize protein crystallization within the cells. A higher MOI does not improve in cellulo crystallization, independent of the crystallizing protein. d Visible crystal growth in High Five cells is detectable from 36 h after infection with an MOI of 1 onwards. Growth characteristics are variable but follow a comparable scheme with the maximal fraction of crystal-containing cells obtained between 72 and 96 hpi. The only exception is IMPDH cyto, showing a continuous increase in crystal containing cells. b–d Data of three independent experiments are presented as mean values ± SD.