Disassembly and reassembly improves morphology and thermal stability of human papillomavirus type 16 virus-like particles.
Recombinant human papillomavirus (HPV) 16 L1 protein self-assembles into virus-like particles (VLPs) with diameters of 40 to 60 nm, which are key components in prophylactic HPV vaccines. Marked improvement in morphology and thermal stability on VLP disassembly and reassembly was demonstrated at production scale. Differential scanning calorimetry showed enhanced conformational stability as indicated by the unfolding temperatures and peak heights/areas. Cloud point studies indicated (1) a much lower propensity for post-reassembly VLPs to aggregate during a time course study and (2) much higher cloud point temperatures. In-solution atomic force microscopy showed more uniform size distribution and fully closed particles, with evidence of virion-like assembly revealed by the structural details from a single particle image. Similar approaches for the reassembly of other recombinant VLPs with intrinsic conformational switches would be expected to improve the particle properties and render nanoparticles more suitable for use as vaccines or therapeutics.


BACKGROUND
The authors of this study demonstrated that recombinant human papillomavirus 16 L1 protein self-assembles into virus-like particles (VLPs) with marked improvement in morphology and thermal stability on VLP disassembly and reassembly at production scale. This is expected to render these nanoparticles more suitable for use as vaccines or therapeutics.
Merck Research Laboratories, Merck & Co., West Point, Pennsylvania, USA. qinjianzhao@gmail.com
Qinjian
Q
Zhao
Michael J
MJ
Allen
Yang
Y
Wang
Bei
B
Wang
Ning
N
Wang
Li
L
Shi
Robert D
RD
Sitrin
D002152:Calorimetry, Differential Scanning; D052162:Human papillomavirus 16 / Q000737:chemistry* / Q000648:ultrastructure; D006801:Humans; D018625:Microscopy, Atomic Force; D030361:Papillomavirus Infections / Q000517:prevention & control / Q000821:virology; D053918:Papillomavirus Vaccines / Q000737:chemistry*; D011487:Protein Conformation; D055550:Protein Stability; D058767:Protein Unfolding; D011994:Recombinant Proteins / Q000737:chemistry; D013696:Temperature; D014764:Viral Proteins / Q000737:chemistry* / Q000648:ultrastructure; D014771:Virion / Q000737:chemistry* / Q000648:ultrastructure
D053918:Papillomavirus Vaccines; D011994:Recombinant Proteins; D014764:Viral Proteins