Fig. 1: Construction of MPXV protein VLPs. | Nature Communications

Fig. 1: Construction of MPXV protein VLPs.

From: Mpox multiprotein virus-like nanoparticle vaccine induces neutralizing and protective antibodies in mice and non-human primates

Fig. 1

a MPXV clade Ia M1, B6, and A35 proteins were modified for secretion and conjugated to a SpyCatcher scaffold (SC-mi3) with 60 attachment sites. b Secreted M1, A35, and B6 proteins fused to SpyTag were purified from the medium of transfected cells by Ni-NTA chromatography. Purified proteins were treated with SDS with (left) or without (right) a reducing agent and analyzed by SDS polyacrylamide gel electrophoresis. A Western blot probed with an antibody to histidine tag, representative of five independent purification experiments, is shown. The left lane labeled M shows marker proteins with their mass indicated in kDa. c Conjugation of M1 protein to SC-mi3. Purified SpyTagged M1 was conjugated to 2 µM SC-mi3 at indicated ratios and analyzed by SDS gel electrophoresis. A Coomassie blue-stained gel is shown. Marker proteins (M) at left, ratios of SC-mi3 to M1 at top, positions of SC-mi3-M1 conjugate, SC-mi3, and M1 on right. Similar conjugation results with 5:1 ratios were obtained in five separate experiments with M1, A35 and B6. d Coomassie blue-stained SDS gel analysis of SC-mi3 conjugates derived from purified VLPs used for immunization of mice. Similar results were obtained for five separate purifications.

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