The stabilisation of purified, reconstituted P-glycoprotein by freeze drying with disaccharides

Adam Heikal, Karl Box, Alice Rothnie, Janet Storm, Richard Callaghan, Marcus Allen

Research output: Contribution to journalArticlepeer-review


The drug efflux pump P-glycoprotein (P-gp) (ABCB1) confers multidrug resistance, a major cause of failure in the chemotherapy of tumours, exacerbated by a shortage of potent and selective inhibitors. A high throughput assay using purified P-gp to screen and characterise potential inhibitors would greatly accelerate their development. However, long-term stability of purified reconstituted ABCB1 can only be reliably achieved with storage at -80 °C. For example, at 20 °C, the activity of ABCB1 was abrogated with a half-life of <1 day. The aim of this investigation was to stabilise purified, reconstituted ABCB1 to enable storage at higher temperatures and thereby enable design of a high throughput assay system. The ABCB1 purification procedure was optimised to allow successful freeze drying by substitution of glycerol with the disaccharides trehalose or maltose. Addition of disaccharides resulted in ATPase activity being retained immediately following lyophilisation with no significant difference between the two disaccharides. However, during storage trehalose preserved ATPase activity for several months regardless of the temperature (e.g. 60% retention at 150 days), whereas ATPase activity in maltose purified P-gp was affected by both storage time and temperature. The data provide an effective mechanism for the production of resilient purified, reconstituted ABCB1.
Original languageEnglish
Pages (from-to)37-44
Number of pages8
Issue number1
Early online date17 Oct 2008
Publication statusPublished - Feb 2009

Bibliographical note

© 2009, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International


  • adenosine triphosphatases
  • animals
  • differential scanning calorimetry
  • disaccharides
  • freeze drying
  • glycerol
  • humans
  • maltose
  • moths
  • p-glycoprotein
  • phase transitions
  • proteolipids
  • recombinant proteins
  • temperature
  • trehalose
  • ABCB1
  • multidrug resistance
  • lyophilisation
  • high throughput
  • liposome
  • ATPase
  • proteoliposomes
  • chemotherapy
  • purification


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