Protocol for removal of alum from protein (eg. vaccine)

5 posts / 0 new
Last post
Aggi
Aggi's picture
Protocol for removal of alum from protein (eg. vaccine)

Hi all,

I'm working with HBsAg protein and I need a protocol to remove the adsorbed alum in commercial vaccines to retrieve the HBsAg.

I have tried using tri-sodium citrate to bind the alum, but I get no intact protein in the supernatant.

Can anyone help?

Thank you

Fraser Moss
Fraser Moss's picture
take a look at these

take a look at these references

J Chromatogr A 2005, 1095, 119-25: "Ion-exchange chromatography of hepatitis B virus surface antigen from a recombinant Chinese hamster ovary cell line"

Vaccine 1997, 15, 637-47: "Comparison between hepatitis B surface antigen (HBsAg) particles derived from mammalian cells (CHO) and yeast cells (Hansenula polymorpha): composition, structure and immunogenicity"

Tirumal Rao
Tirumal Rao's picture
Hellow,

Hellow,
The ability of the Pichia pastoris-based technology for large-scale production of recombinant hepatitis B virus surface antigen (HBsAg) and both reproducibly purify HBsAg and remove most of the relevant contaminants was ascertained by evaluating ten industrial production batches, five in 1993 and five in 1998. At an early stage, the clarification of mechanically disrupted yeast cells by acid precipitation renders HBsAg with a purity as low as 3.8±0.6%. However, by adsorption/desorption from diatomaceous earth matrix, the purity of HBsAg rapidly increases to 18.8±5%, which is suitable for chromatographic processing. This step also eliminates non-particulated forms of HBsAg, significantly lowers the amount of carbohydrates and lipids, and concentrates the HBsAg 4.8-fold. Finally, a sequential purification procedure that includes large-scale immunoaffinity, ion-exchange, and size-exclusion chromatographies further purifies the preparation, resulting in a product (HBsAg at a concentration of 1.3±0.2 g l-1) with a purity of 95% or more. Furthermore, each of the other contaminants measured reaches the following low levels per 20 µg HBsAg: host deoxyribonucleic acid (<10 pg), carbohydrates (1.2±0.02 µg), lipids (14±0.28 µg), immunopurification-released immunoglobulin G (less than 100 ppm), and endotoxins (106.7±19.3 pg). These values are below those specified for recombinant DNA hepatitis B vaccines according to World Health Organization (WHO) guidelines.

Tirumal

ncruzg
ncruzg's picture
I'm working with HBsAg

I'm working with HBsAg protein and I need a protocol to remove the adsorbed alum in commercial vaccines to retrieve the HBsAg.

The FFM
The FFM's picture
Please click the link below

Please click the link below to take a look in our Protocols section at this method

Elution of HBsAg from aluminum hydroxide adjuvant

Derived from Iyer et al. Vaccine Volume 22, Issues 11-12, 29 March 2004, Pages 1475-1479