Accession Number : AD1013906


Title :   Purification and Characterization of Recombinant Vaccinia L1R Protein from Escherichia coli


Descriptive Note : Technical Report,01 Apr 2015,01 Jan 2016


Corporate Author : Excet, Inc Gunpowder United States


Personal Author(s) : Cole,Stephanie ; Blum,Steven ; Maciel,Jorge ; Miklos,Aleksandr ; Dixon,Melissa ; Funk,Vanessa


Full Text : https://apps.dtic.mil/dtic/tr/fulltext/u2/1013906.pdf


Report Date : 01 Aug 2016


Pagination or Media Count : 22


Abstract : The Vaccinia virus is closely related to the causative agent of smallpox and as such, is frequently used as a surrogate for smallpox in research and development efforts. The Vaccinia surface protein, L1R, is critical for viral entry into host cells, and because of its close similarity to its smallpox counterpart, it is a logical target for therapeutic and vaccine development. In this report, we describe a procedure and the typical outcome for the recombinant expression and purification of the L1R protein in Escherichia coli cells. L1R is solubilized and refolded from inclusion bodies after induction in E. coli, and its purification proceeds through two rounds of immobilized metal affinity chromatography, followed by size-exclusion chromatography. The typical yield is approximately 2 mg/L of bacterial culture. The recombinant L1R was shown to bind to a known anti-L1Rantibody, which indicated that the protein was intact and properly folded. This procedure allows for the in-house production o fL1R for research efforts such as antibody discovery and immunoassay development.


Descriptors :   VACCINIA VIRUS , vaccines , SMALLPOX VIRUS , ESCHERICHIA COLI , proteins , PURIFICATION , CHROMATOGRAPHY , ANTIGENS , HOSTS BIOLOGY , RECOMBINANT DNA , GENOME , IMMUNOASSAY , CULTURES BIOLOGY


Distribution Statement : APPROVED FOR PUBLIC RELEASE