Principles of Cell Disruption in Biopharmaceuticals

Mechanical methods: These disrupt cell structures through mechanical forces such as compression and shearing. The underlying principle involves using external physical force to directly rupture cell walls and membranes. Common methods include high-pressure homogenization (forcing a cell suspension through a narrow gap under high pressure to generate high-speed shear forces), bead milling (using high-speed agitation of grinding beads to generate shear and impact forces), and ultrasonic disruption (utilizing the ultrasonic cavitation effect to create pressure differentials that rupture cells).

Non-mechanical methods:

(1) Chemical methods: Utilizing surfactants (e.g., Triton X-100) or organic solvents (e.g., ethanol) to disrupt the lipid bilayer structure of the cell membrane, thereby increasing membrane permeability;

(2) Enzymatic methods: Employing hydrolases-such as lysozyme (which breaks down peptidoglycan in bacterial cell walls) or cellulase (which breaks down cellulose in plant cell walls)-to specifically degrade components of the cell wall or membrane;

(3) Physical methods:
① Freeze-thaw method: Repeated freezing and thawing causes intracellular water to form ice crystals; the resulting volumetric expansion leads to cell rupture;
② Osmotic shock method: Cells are transferred from a hypertonic environment to a hypotonic environment, causing them to absorb water, swell, and rupture.