Butyrylcholinesterase formulated in liposomes

Ilana Schumacher, Aya Arad, Rimona Margalit*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Exogenous cholinesterases have the potential to take part in defence against organophosphate toxins, by acting as scavenger systems. Postulating that formulation in liposomes could enhance the toxin-scavenging potential of these enzymes, we have initiated studies of such formulations and are reporting here our first steps, exploring butyrylcholinesterase (BChE) in multi-lamellar liposomes composed of phosphatidylcholine. We started by developing an essential research tool: a multisample, sensitive and rapid enzyme-activity assay, based on the Ellman reaction, that could be performed directly on liposome-containing samples. Using an ELISA reader equipped to follow time-dependant absorbency changes, 10 min sufficed to assay 96 samples simultaneously. Next, several key properties of liposome-formulated BChE were explored and the major findings were: (i) the encapsulated enzyme was found to retain its activity. (ii) Enzyme activity was found to increase (at constant enzyme concentration) in the presence of the lipid, in a lipid-concentration dependant manner. Through data analysis it was possible to attribute this effect to changes in k(cat). (iii) Good, reproducible, encapsulation efficiencies (for macromolecules) in the range of 30% were obtained at liposome concentrations of 100 mM lipid. (iv) Free BChE was completely susceptible to proteolysis under conditions mimicking enzymically-hostile biological environments, whereas ≥ 60% of the liposome-formulated BChE was protected, found to be inaccessible to the proteolytic enzymes. (v) Short-term exposures of free and liposome-encapsulated BChE to the inhibitor paraoxon, generated significant losses in enzyme activity. Residual activities of both BChE formulations dropped considerably over the paraoxon concentration range of 0.02-0.11 μM, down to 3 and 11% for free and liposome-encapsulated enzyme respectively. These data are a clear indication that the encapsulated BChE was accessible to the inhibitor, indicating that such liposomal formulations have the potential to perform as the desired scavenger systems.

Original languageEnglish
Pages (from-to)225-230
Number of pages6
JournalBiotechnology and Applied Biochemistry
Issue number3
StatePublished - 1999


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