The human papillomavirus vaccine is based on recombinant L1 protein produced in yeast or insect cells. L1 is a major capsid protein that self-assembles into icosahedral virus-like particles (VLP). Conventionally, ultracentrifugation using a sucrose cushions or cesium chloride density gradients, and size-exclusion chromatography, have been used for small scale purification of L1 protein. However, these methods require a great deal of time and labor, and are not suitable for industrial-scale purification. To resolve these problems, simple one-step chromatography method has been developed for purifying recombinant HPV16 L1 protein produced in Pichia pastoris. Majority percentage of the contaminating protein was removed by ammonium sulfate precipitation and by precipitating contaminants prior to the chromatography step. Cation-exchange chromatography method has been developed using two different resins with considerably high protein recovery. In addition, it was confirmed that purified HPV16 L1 protein  self-assembles into VLP by TEM and SEC MALS. The one-step chromatography method proposed here can potentially reduce the time, cost and labor needed for purification of L1 protein, and facilitate the study of HPV vaccines.

One-step chromatographic purification of human papillomavirus type 16 L1 protein from Pichia pastoris

ACS Spring 2022

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