A preparative purification process for recombinant Hepatitis B core antigen using online capture by expanded bed adsorption followed by size-exclusion chromatography

• Main Authors: Ho, Chin Woi, Tan, Wen Siang, Chong, Fui Chin, Ling, Tau Chuan, Tey, Beng Ti
• Format: Article
• Language: English
• Published: Korean Society for Microbiology and Biotechnology 2009
• Online Access: http://psasir.upm.edu.my/id/eprint/12790/1/A%20preparative%20purification%20process%20for%20recombinant%20Hepatitis%20B%20core%20antigen%20using%20online%20capture%20by%20expanded%20bed%20adsorption%20followed%20by%20size.pdf; http://psasir.upm.edu.my/id/eprint/12790/; http://www.jmb.or.kr/journal/viewJournal.html?year=2009&vol=19&num=4&page=416

Hepatitis B core antigen (HBcAg) is an important serological marker used in the diagnosis of hepatitis B virus (HBV) infections. In the current study, a fast and efficient preparative purification protocol for truncated HBcAg from Escherichia coli disruptate was developed. The recombinant HBcAg was first captured by anion exchange expanded bed adsorption chromatography integrated with a cell disruption process. This online capture process has shortened the process time and eliminated the “hold-up” period that may be detrimental to the quality of target protein. The eluted product from the expanded bed adsorption chromatography was subsequently purified using size-exclusion chromatography. The results showed that this novel purification protocol achieved a recovery yield of 45.1% with a product purity of 88.2%, which corresponds to a purification factor of 4.5. The recovered HBcAg is still biologically active as shown by ELISA test.