Isolation and purification of recombinant proteins, antibodies and plasmid DNA with hydroxyapatite chromatography

Title data

Hilbrig, Frank ; Freitag, Ruth:
Isolation and purification of recombinant proteins, antibodies and plasmid DNA with hydroxyapatite chromatography.
In: Biotechnology Journal. Vol. 7 (2012) Issue 1 . - pp. 90-102.
ISSN 1860-7314
DOI: https://doi.org/10.1002/biot.201100015

Official URL:

Abstract in another language

Hydroxyapatite and related stationary phases increasingly play a role in the downstream processing of high-value biological materials, such as recombinant proteins, therapeutic antibodies and pharmaceutical-grade plasmid DNA. Chromatographic hydroxyapatite is an inorganic, ceramic material identical in composition, if not in structure, to calcium phosphate found in human bones and teeth. The interaction of hydroxyapatite with biomacromolecules is complex and highly dynamic, which can make predicting performance difficult, but also allows the design of very selective isolation processes. This review discusses the currently commercially available chromatographic materials, different retention mechanisms supported by these materials and differential exploitation for the design of highly specific isolation procedures. The state of the art of antibody purification by hydroxy- and fluoroapatite is reviewed together with tested routines for method development and implementation. Finally, the isolation of plasmid DNA is discussed, since the purification of DNA therapeutics at a sufficiently large scale is an emerging need in bioprocess development and perhaps the area in bioseparation where apatite chromatography can make its most important contribution to date.

Further data

Item Type:	Article in a journal
Refereed:	Yes
Keywords:	Affinity chromatography; Apatite; Downstream processing; Mixed-mode interaction; Retention mechanism
Institutions of the University:	Faculties > Faculty of Engineering Science Faculties > Faculty of Engineering Science > Chair Process Biotechnology Faculties > Faculty of Engineering Science > Chair Process Biotechnology > Chair Process Biotechnology - Univ.-Prof. Dr. Ruth Freitag Faculties
Result of work at the UBT:	Yes
DDC Subjects:	600 Technology, medicine, applied sciences 600 Technology, medicine, applied sciences > 620 Engineering
Date Deposited:	23 Feb 2016 15:11
Last Modified:	26 Apr 2022 11:21
URI:	https://eref.uni-bayreuth.de/id/eprint/31024