The recovery of the hepatitis B virus surface antigen (HBsAg) from a recombinant P. pastoris strain disruption and precipitation studies
1993; Wiley; Volume: 13; Issue: 2 Linguagem: Inglês
10.1002/abio.370130207
ISSN1521-3846
AutoresRolando Páez, Alberto Agraz, Luís Herrera,
Tópico(s)Toxin Mechanisms and Immunotoxins
ResumoAbstract Cell disruption studies for the extraction of HBsAg from a recombinant P. pastoris strain (r‐HBsAg) were done using a bead mill disintegrator. Three sequential passages (4 min retention time each) were enough to disrupt the cells and extract most of the r‐HBsAg and soluble proteins. An acid precipitation step was performed just after cell disruption to precipitate proteins together with the cell debris. Different precipitation pH values (2.5 to 6.0) were investigated. A pH value of 4.2 was selected as a compromise between recovery and improvement of specific activity. A 6 to 8‐fold enhancement of the specific activity was obtained, having a r‐HBsAg overall yield of about 80%. The influencing presence of a chaotropic salt (potassium thiocyanate) during the acid precipitation step was also studied.
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