Detergents for Cell Rupture
Detergents (or surfactants) are used in cell lysis solutions because they disrupt the distinct interface between hydrophobic and hydrophilic systems. They help to solubilize membrane proteins and lipids, thereby causing the cell to lyse and release its contents.
Detergents are comprised of a polar hydrophilic head group and a nonpolar hydrophobic tail. They are categorized by the nature of the head group as either ionic, nonionic or zwitterionic.
Nonionic and zwitterionic detergents are generally milder and less denaturing than ionic detergents. Examples include CHAPS (zwitterionic) and Triton (nonionic). Ionic detergents are considered to be harsh detergents. They are strong solubilizing agents and tend to denature proteins. Examples include soap or alcohol ethoxysulfates (anionic detergents), and quaternary ammonium compounds (cationic detergents).
Physical Methods of Cell Rupture
There are several methods that are commonly used to physically lyse cells, including:
- Mechanical disruption: Using various equipment to cut, chop, grind and crush the sample.
- Sonication: Using pulsed, high frequency sound waves to lyse cells. This process can be direct (a probe is inserted into the sample) or indirect (the energy is transmitted through a bath of water into the sample vessels).
- Freeze-Thaw method: This technique involves freezing a cell suspension and then thawing the material at room temperature. This causes cells to swell and ultimately break as ice crystals form during the freezing process and then contract during thawing. The process is repeated as necessary.
- Homogenization: The sample is forced through a very narrow nozzle. The higher the amount of energy applied during the homogenization process, the more efficient the cell lysis.
Which one to choose?
Detergent-based lysis is a popular method for cell rupture. It is fairly easy to do and does not require any special equipment. However, detergent cell lysis is often too mild, and needs to be done in conjunction with a physical method like grinding or homogenization. Also, bear in mind that harsh detergents can often damage or destroy the contents of the cell if used incorrectly.
Our high pressure homogenizing technology at BEE International allows you to gently rupture cells without damaging the valuable intracellular materials. No harsh chemicals are introduced into the process, and all results are 100% scalable to manufacturing.
Still unsure? Download our free eBook: 7 Key Factors to Consider When Choosing a Cell Lysis Method or contact us here.