Screen-less expanded bed column: new approach for the recovery and purification of a malaria transmission blocking vaccine candidate from Pichia pastoris

Abstract

An experimental malaria transmission blocking vaccine antigen, Pfs25H, expressed and secreted from Pichia pastoris was recovered and purified using a screenless expanded bed column equipped with a rotating fluid distribution system. This column was able to accommodate feed stock, containing 30% biomass, at a flow rate of 300-400 cm/h without affecting column stability. This capability is three times higher than the capability of the expanded bed column currently in use, which is equipped with a perforated plate fluid distribution system; this design could accommodate biomass concentrations of only up to 10%. The screen-less design did not affect the binding capacity, purification level or process yield and, therefore, shorten the process. Purified Pfs25H of 6.4 g were recovered from 37 l of Pichia pastoris culture in one step.

Expanded bed adsorption of bromelain (E.C. 3.4.22.33) from Ananas comosus crude extract

“…Expanded Bed Adsorption (EBA) is a chromatographic technique for separation and purification of biological products directly from crude feedstock, without centrifugation, microfiltration and other prior clarification steps (Chase, 1994;Hjorth;Thömmes, 1997;Anspach et al, 1999;Trinh et al, 2006;Camprubi et al, 2006;Tan et al, 2006). This technique permits crude feeding into the chromatographic column without initial treatment and, as the bed expands, it increases adsorbent surface contact, making interaction with the target molecule more effective (Roy et al, 1999;Fernandez-Lahore et al, 2001).…”