A large-scale purification of recombinant histone H1. 5 from Escherichia coli
(2001) In Protein Expression and Purification 23(1). p.38-44- Abstract
- An Escherichia coli expression system has been constructed for production of biologically active recombinant histone H1.5. A process of fermentation and purification method at a large scale has been developed. Recombinant histone H1.5 was released from the high density cultured cells by high-pressure homogenization. For an efficient removal of cell debris and partial purification of basic histone H1.5 in a single step, the whole cell lysates were directly loaded onto an expanded bed column packed with the strong cation exchanger (Streamline SP). Complete removal of various impurities was achieved by a combination of hydroxyapatite chromatography and the following cation exchange chromatography with high grade strong cation exchanger (POROS... (More)
- An Escherichia coli expression system has been constructed for production of biologically active recombinant histone H1.5. A process of fermentation and purification method at a large scale has been developed. Recombinant histone H1.5 was released from the high density cultured cells by high-pressure homogenization. For an efficient removal of cell debris and partial purification of basic histone H1.5 in a single step, the whole cell lysates were directly loaded onto an expanded bed column packed with the strong cation exchanger (Streamline SP). Complete removal of various impurities was achieved by a combination of hydroxyapatite chromatography and the following cation exchange chromatography with high grade strong cation exchanger (POROS 20 HS), and finally endotoxins were removed by ultrafiltration using a 100-kDa cut-off membrane, which gave the level of endotoxin below 0.5 EU/mg. The … (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1f03f840-e5d7-4bda-bf16-dd7de8f9e05b
- author
- Pyo, Sang-Hyun LU ; Lee, Jae-Hyun ; Park, Heung-Bok ; Hong, Seung-Suh and Kim, Jin-Hyun
- publishing date
- 2001-10-01
- type
- Contribution to journal
- publication status
- published
- in
- Protein Expression and Purification
- volume
- 23
- issue
- 1
- pages
- 38 - 44
- publisher
- Academic Press
- ISSN
- 1046-5928
- DOI
- 10.1006/prep.2001.1471
- language
- English
- LU publication?
- no
- id
- 1f03f840-e5d7-4bda-bf16-dd7de8f9e05b
- date added to LUP
- 2025-09-05 17:51:35
- date last changed
- 2025-09-17 12:26:59
@article{1f03f840-e5d7-4bda-bf16-dd7de8f9e05b, abstract = {{An Escherichia coli expression system has been constructed for production of biologically active recombinant histone H1.5. A process of fermentation and purification method at a large scale has been developed. Recombinant histone H1.5 was released from the high density cultured cells by high-pressure homogenization. For an efficient removal of cell debris and partial purification of basic histone H1.5 in a single step, the whole cell lysates were directly loaded onto an expanded bed column packed with the strong cation exchanger (Streamline SP). Complete removal of various impurities was achieved by a combination of hydroxyapatite chromatography and the following cation exchange chromatography with high grade strong cation exchanger (POROS 20 HS), and finally endotoxins were removed by ultrafiltration using a 100-kDa cut-off membrane, which gave the level of endotoxin below 0.5 EU/mg. The …}}, author = {{Pyo, Sang-Hyun and Lee, Jae-Hyun and Park, Heung-Bok and Hong, Seung-Suh and Kim, Jin-Hyun}}, issn = {{1046-5928}}, language = {{eng}}, month = {{10}}, number = {{1}}, pages = {{38--44}}, publisher = {{Academic Press}}, series = {{Protein Expression and Purification}}, title = {{A large-scale purification of recombinant histone H1. 5 from <i>Escherichia coli</i>}}, url = {{http://dx.doi.org/10.1006/prep.2001.1471}}, doi = {{10.1006/prep.2001.1471}}, volume = {{23}}, year = {{2001}}, }