Open AccessPurification of a delayed hypersensitivity‐inducing protein from Listeria monocytogenes
Author(s) -
Antonissen A.C.J.M.,
Lemmens P.J.M.R.,
Bosch J.F.,
Boven C.P.A.
Publication year - 1986
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1986.tb01355.x
Subject(s) - listeria monocytogenes , chromatography , chemistry , fractionation , elution , urea , size exclusion chromatography , delayed hypersensitivity , sepharose , ion chromatography , column chromatography , potassium thiocyanate , antigen , biochemistry , bacteria , biology , inorganic chemistry , genetics , enzyme
Cell‐envelope fragments were prepared from Listeria monocytogenes L242, serotype 4b. Delayed hypersensitivity (DH)‐inducing proteins were extracted with deoxycholate and separated into two fractions by filtration through a Sephacryl S‐200 column equilibrated with deoxycholate buffer. The second peak eluting from the Sephacryl column was fractionated using ion exchange chromatography on a DEAE Sepharose CL‐6B column in the presence of 6 M urea. A purified 20 400‐Da protein which induced DH against L. monocytogenes was obtained by isocratic elution. Three other DH‐inducing fractions containing several protein bands were eluted by a gradient of potassium thiocyanate (KSCN) in urea buffer. Our results indicate that denaturing conditions can be employed for the fractionation and purification of DH inducing proteins from L. monocytogenes . In addition, it is suggested that the procedure described might also be useful for the purification of other antigens involved in cellular immune reactions.