Inderscience Publishers

Immobilisation of HEP-1 by covalent attachment to aldehyde–agarose gels

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A procedure of covalent multipoint attachment has been performed for immobilisation of HEP-1 using two different pH conditions. This study revealed that in more alkaline media as pH 10 the antibody immobilises quickly. In this condition, supports are able to bind 4 mg of HEP-1 per mL of gel. In order to test the activity of this new immobilised derivative, it was used in the affinity purification of the recombinant Hepatitis B surface antigen. We made a comparison between immunosorbents activated by the CNBr and aldehyde–agarose methods and subsequently studied the recovery during 23 cycles. The amounts of eluted antigen per millilitre decrease 1.5 times with the increment of the cycles either aldehyde–agarose or CNBr gels. We also measured the leakage in each cycle and it was below 15 ng IgG/mL.

Keywords: HEP 1, r-HBsAg, aldehyde–agarose gels, covalent multipoint attachment, immobilisation

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