A method for the purification of bacterial flagellin that allows simple upscaling

HIRIART YANINA,; ERREA AGUSTINA; GONZÁLEZ MACIEL, DOLORES; M RUMBO,

WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY

SPRINGER

Lugar: Berlin; Año: 2012 vol. 2012 p. 15 - 21

0959-3993

Due to putative applications as adjuvant, immunomodulatory agent or vaccinal antigen, there is a growing interest in enterobacterial flagellins, that may result in a demand on producing flagellin at industrial scale. Traditionally flagellin has been produced at laboratory scale by flagellum shearing of bacterial surface and subsequent ultracentrifugation. The main drawback of these methods is the need to use low agitation culture to avoid loss of flagellum due to shearing during culture. In the present work we describe the use of a different protocol to produce flagellin with higher yield than traditional laboratory scale protocols. Among several alternatives tested, we selected the use of cross flow filtration to concentrate bacterial culture. This procedure combines extensive shearing that takes place during concentration with volume reduction, greatly simplifying the downstream processing. On the other hand, it allows the use of highly agitated culture condition, since any sheared flagellin is retained in the bacterial concentrate. Flagellin obtained following this procedure showed similar innate activating capacity either in vivo or in vitro, than lab-scale produce flagellin. The procedure developed is very flexible to increase in the production scale, enhances the yield in mg of flagellin/L of culture and does not require expensive equipment.