Methods for sonochemical manufacture of proteinaceous ultrasound contrast agents

Laboratory procedures for the sonochemical manufacture of proteinaceous contrast agents are reviewed and documented. The objective was to develop a standard protocol to enable ultrasound contrast researchers to develop consistent agents. Proteinaceous agents continue to be in clincal use and are the subject of renewed interest owing to their potential as a platform for antibody-targeted diagnostics. The agents are prepared by sonication. Protocols for agent preparation are scattered throughout literature with methodologies varying between laboratories. While new imaging technologies improve agent visualisation, there are limited correlations between contrast agent manufacturing methods and in vitro performance. The present experimental study has determined that successful production of a stable shell coating the bubble is a result of precise attention to denaturation of protein prior to sonication. The resulting shell coats the bubble and greatly slows down gas dissolution that would otherwise result in rapid disappearance of the agent or poor shelf life. The fluid-dynamical process by which the agent is formed is discussed. Issues of technique such as the location of the sonotrode tip, sonication time and power, and and the container size are detailed. Finally, procedures for quantitative characterisation of the agent by a range of instruments are discussed. In conclusion, a complete and consistent protocol from the initial biochemical source to the agent characterisation can be produced

Contemporary Developments in Water Law

Center for Water and the Environmen