Purification of rabbit polyclonal immunoglobulin G with ammonium sulphate precipitation and mixed-mode chromatography
Immunoglobulins G (IgG) against hepatitis B core antigen (HBcAg) was successfully purified using a purification scheme comprising ammonium sulphate precipitation and SepFast™ MM AH-1 column chromatography. Ammonium sulphate precipitation performed at 40% saturation was optimum in terms of the recove...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2015
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| Online Access: | http://psasir.upm.edu.my/id/eprint/46050/ http://psasir.upm.edu.my/id/eprint/46050/1/Purification%20of%20rabbit%20polyclonal%20immunoglobulin%20G%20with%20ammonium%20sulphate%20precipitation%20and%20mixed-mode%20chromatography.pdf |
| _version_ | 1848850585210585088 |
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| author | Mariam, S. H. S Ooi, Chien Wei Tan, Wen Siang Abdullah, Janna Ong Ariff, Arbakariya Tey, Beng Ti |
| author_facet | Mariam, S. H. S Ooi, Chien Wei Tan, Wen Siang Abdullah, Janna Ong Ariff, Arbakariya Tey, Beng Ti |
| author_sort | Mariam, S. H. S |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Immunoglobulins G (IgG) against hepatitis B core antigen (HBcAg) was successfully purified using a purification scheme comprising ammonium sulphate precipitation and SepFast™ MM AH-1 column chromatography. Ammonium sulphate precipitation performed at 40% saturation was optimum in terms of the recovered polyclonal IgG concentration (7.8 mg/ml) and the removal of albumin (72%). The yield, purity and purification factor achieved from this simple purification method were 99%, 94% and 7.8, respectively. The IgG recovered from ammonium sulphate precipitation was subjected to SepFast™ MM AH-1 column chromatography and the purity of IgG was further increased to 98%, corresponding to a purification factor of 8.1. Protein aggregation was also reduced significantly in the purified IgG sample. Furthermore, the salt content in the purified sample was reduced by 75% and therefore the need of desalting final product was eliminated. Enzyme-linked immunosorbent assay (ELISA) showed that the antigenicity of anti-HBcAg IgG obtained after these purification processes was maintained. |
| first_indexed | 2025-11-15T10:08:37Z |
| format | Article |
| id | upm-46050 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T10:08:37Z |
| publishDate | 2015 |
| publisher | Elsevier |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-460502022-03-28T07:12:57Z http://psasir.upm.edu.my/id/eprint/46050/ Purification of rabbit polyclonal immunoglobulin G with ammonium sulphate precipitation and mixed-mode chromatography Mariam, S. H. S Ooi, Chien Wei Tan, Wen Siang Abdullah, Janna Ong Ariff, Arbakariya Tey, Beng Ti Immunoglobulins G (IgG) against hepatitis B core antigen (HBcAg) was successfully purified using a purification scheme comprising ammonium sulphate precipitation and SepFast™ MM AH-1 column chromatography. Ammonium sulphate precipitation performed at 40% saturation was optimum in terms of the recovered polyclonal IgG concentration (7.8 mg/ml) and the removal of albumin (72%). The yield, purity and purification factor achieved from this simple purification method were 99%, 94% and 7.8, respectively. The IgG recovered from ammonium sulphate precipitation was subjected to SepFast™ MM AH-1 column chromatography and the purity of IgG was further increased to 98%, corresponding to a purification factor of 8.1. Protein aggregation was also reduced significantly in the purified IgG sample. Furthermore, the salt content in the purified sample was reduced by 75% and therefore the need of desalting final product was eliminated. Enzyme-linked immunosorbent assay (ELISA) showed that the antigenicity of anti-HBcAg IgG obtained after these purification processes was maintained. Elsevier 2015 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/46050/1/Purification%20of%20rabbit%20polyclonal%20immunoglobulin%20G%20with%20ammonium%20sulphate%20precipitation%20and%20mixed-mode%20chromatography.pdf Mariam, S. H. S and Ooi, Chien Wei and Tan, Wen Siang and Abdullah, Janna Ong and Ariff, Arbakariya and Tey, Beng Ti (2015) Purification of rabbit polyclonal immunoglobulin G with ammonium sulphate precipitation and mixed-mode chromatography. Separation and Purification Technology, 144. pp. 133-138. ISSN 1383-5866; ESSN: 1873-3794 https://www.sciencedirect.com/science/article/pii/S1383586615000969 10.1016/j.seppur.2015.02.012 |
| spellingShingle | Mariam, S. H. S Ooi, Chien Wei Tan, Wen Siang Abdullah, Janna Ong Ariff, Arbakariya Tey, Beng Ti Purification of rabbit polyclonal immunoglobulin G with ammonium sulphate precipitation and mixed-mode chromatography |
| title | Purification of rabbit polyclonal immunoglobulin G with ammonium sulphate precipitation and mixed-mode chromatography |
| title_full | Purification of rabbit polyclonal immunoglobulin G with ammonium sulphate precipitation and mixed-mode chromatography |
| title_fullStr | Purification of rabbit polyclonal immunoglobulin G with ammonium sulphate precipitation and mixed-mode chromatography |
| title_full_unstemmed | Purification of rabbit polyclonal immunoglobulin G with ammonium sulphate precipitation and mixed-mode chromatography |
| title_short | Purification of rabbit polyclonal immunoglobulin G with ammonium sulphate precipitation and mixed-mode chromatography |
| title_sort | purification of rabbit polyclonal immunoglobulin g with ammonium sulphate precipitation and mixed-mode chromatography |
| url | http://psasir.upm.edu.my/id/eprint/46050/ http://psasir.upm.edu.my/id/eprint/46050/ http://psasir.upm.edu.my/id/eprint/46050/ http://psasir.upm.edu.my/id/eprint/46050/1/Purification%20of%20rabbit%20polyclonal%20immunoglobulin%20G%20with%20ammonium%20sulphate%20precipitation%20and%20mixed-mode%20chromatography.pdf |