Cell rupture, a technique used in labs across the world, allows for extraction of intracellular molecules via one of many processes. Chemical and mechanical disruption are two of the more commonly used classes of methods, where enzyme treatment may be employed as a chemical, and high pressure homogenization as a mechanical, procedure. Each may confer significant benefits on the condition of an appropriate match between the researcher’s goal, sample composition, and process capability. Below is an overview of both enzyme and high pressure homogenization procedures in relation to cell rupture, and an analysis on which may be more appropriate for your purposes.
Overview of Processes
Enzyme treatment can be an effective avenue in terms of breaking down cell walls, removing unwanted contaminants, generate protoplasts, and promote DNA isolation by breaking down DNA-binding proteins. The type of enzyme used will depend on its intended use; for example, proteases like trypsin and collagenase can release individual cells from tissue, while cellulases can yield protoplasts from plant cells. One of the major benefits of enzymes is that they can target specific cells and/or molecules without leaving contaminants and other unwanted material in the lysate. (1) High pressure homogenization, in contrast, forces a sample through a narrow space while imparting high amounts of force (e.g. pressure, shear, cavitation, turbulence) on the sample. This allows for effective rupture, particularly among cells whose intracellular components are protected by tough cell walls.
Which Technique Should I Use?
To determine which method will be best for your purposes, start off by identifying three factors about your sample: 1) What type of cell it will be comprised of, 2) The intracellular component(s) of interest, and 3) The process volume. The cell type will determine whether a tough cell wall or thin membrane needs to be penetrated to access the molecule of interest. Additionally, knowledge about the molecule of interest like whether it is secreted from the cell and which enzymes can degrade it will hugely influence your machine selection. Finally, while enzymes are appropriate for small process volumes, high pressure homogenizers are more appropriate for medium and large volumes. Analysis of these three factors as a whole will help you make a more informed and accurate selection.
Pion: The Homogenizer Advantage
Use of high quality cell lysis equipment will provide numerous benefits for both production laboratories and consumers alike. And there are plenty of companies on the market to select your equipment from. However, the lysate can be of higher quality and more even consistency when run through top-shelf equipment, most frequently in the form of a high pressure homogenizer. Pion's BEE brand technology is trusted by researchers and lab managers around the world. We deliver an array of key benefits, such as production of nano/micro emulsions and dispersions and lipids and suspensions; these can be used for applications such as injectables, targeted drug delivery, inhalants, time release, anesthetics, and importantly, vaccinations.
In addition, we have extensive experience in the challenges that our customers face as they transition from concept, through to R&D, clinical trials, all-important FDA approval and finally, to manufacturing.
See how Pion can help your cell lysis application by contacting us today.