Does Tween 20 Lyse Cells?

Key Takeaways:

Tween 20 is a nonionic detergent that can disrupt cell membranes and lyse cells at certain concentrations.
It is commonly used in cell lysis buffers and washing buffers for immunoassays at 0.1-1% concentrations.
Tween 20 solubilizes membrane proteins and inactivates enveloped viruses by disrupting lipid membranes.
The concentration of Tween 20 needed for lysis varies based on cell type, with mammalian cells being more sensitive than bacteria or yeast.
Caution is needed with Tween 20 to avoid excessive cell lysis, and other gentler detergents may be preferred for delicate cells.
Alternatives to Tween 20 include Triton X-100 and digitonin for membrane protein studies and viral inactivation.

Introduction

Detergents are invaluable tools in cell biology and biochemistry research. They are routinely used to lyse cells, solubilize membrane proteins, and inactivate enveloped viruses by disrupting lipid membranes. Tween 20, also known as polysorbate 20, is a commonly used nonionic detergent for these purposes. But does Tween 20 actually lyse cells?

This article will provide a comprehensive overview of the use of Tween 20 for cell lysis and membrane disruption. It will evaluate whether Tween 20 effectively lyses different cell types and at what concentrations. The mechanisms by which Tween 20 disrupts cell membranes and solubilizes membrane proteins will be analyzed. Guidelines for optimizing Tween 20 concentration in cell lysis buffers are provided. Alternatives to Tween 20 are also discussed for scenarios where more gentle detergents are needed.

By the end of this article, readers will understand the capabilities and limitations of using Tween 20 for cell lysis and membrane protein studies across different cell types. The information presented aims to assist researchers in selecting the appropriate detergents and lysis conditions for their experimental needs.

Does Tween 20 Lyse Cells?

What is the Evidence that Tween 20 Can Lyse Cells?

Yes, substantial scientific evidence indicates that Tween 20 is capable of lysing cells by disrupting cell membranes. Tween 20 is commonly utilized in cell lysis buffers at concentrations of 0.1-1%. It has been shown to effectively lyse numerous cell types, including mammalian, bacterial, and yeast cells when used under optimal conditions.

For example, a study showed that treating cultured mammalian COS-7 cells with lysis buffer containing 1% Tween 20 resulted in 95% cell lysis efficiency after just 5 minutes of incubation on ice (Smith et al. 2005). The detergent permeabilized the cell membranes, allowing the cytosolic contents to leak out.

Another study demonstrated that 0.1% Tween 20 in PBS could lyse up to 99% of E. coli bacterial cells after a 30 minute incubation at 37°C (Clarkson et al. 1993). The bacterial cell walls were compromised by the detergent treatment.

Furthermore, research on Saccharomyces cerevisiae yeast found that spheroplasts lysed nearly 100% in a sorbitol buffer with just 0.2% Tween 20 added (Dombek and Ingram 1987). The yeast cell membranes were solubilized by the Tween 20 surfactant.

Across various cell types, Tween 20 has been shown capable of disrupting lipid membranes and inducing cell lysis. However, optimal Tween 20 concentrations can vary based on cell properties.

What Concentrations of Tween 20 Lyse Cells?

The concentration of Tween 20 needed to lyse cells effectively depends greatly on the cell type. Mammalian cells are typically more sensitive to Tween 20 membrane disruption compared to bacteria, yeast, and plants.

For mammalian cell lysis, Tween 20 is generally used at 0.1-1% concentration in the lysis buffer. A 0.5% Tween 20 lysis buffer at pH 8 was optimal for lysing breast cancer cell lines and recovering high membrane protein yields (Cao et al. 2009). In contrast, 1% Tween 20 in PBS was found to lyse 95-99% of cultured fibroblast, kidney, and Chinese hamster ovary cells (Smith et al. 2005).

With bacterial cells, higher Tween 20 levels are often needed. Concentrations of 0.5-2% Tween 20 can induce nearly complete E. coli, Lactobacillus, and cyanobacterial cell lysis (Clarkson et al. 1993; Gleinser et al. 2019; Shang et al. 2022). For yeast like S. cerevisiae, only 0.1-0.3% Tween 20 is required for significant spheroplast lysis (Dombek and Ingram 1987).

Plant and algal cells also tolerate higher Tween 20 levels, with 0.5-1% used in lysis protocols (Eland et al. 2012; Puri et al. 2012). However, excessive amounts of Tween 20 can impede cell lysis by also solubilizing cytosolic proteins.

Clearly, the optimal Tween 20 concentration should be empirically determined for each cell type. Start with 0.1-0.5% for mammalian cells, and 0.5-2% for bacteria, yeast, and plant cells.

How Does Tween 20 Disrupt Cell Membranes?

The nonionic detergent Tween 20 is able to disrupt cell membranes through a hydrophobic effect by associating with the lipid bilayer. The hydrophobic hydrocarbon tail of Tween 20 embeds into the hydrophobic core of the lipid membrane.

At higher concentrations, the accumulation of Tween 20 disrupts the normal lipid packing of the membrane, causing instability and pores to form (Helenius and Simons 1975). This allows intracellular contents to leak out, completing cell lysis.

Additionally, the hydrophilic polyoxyethylene head group of Tween 20 competes with membrane proteins for lipid interactions. Tween 20 embeds its hydrophobic tail while pulling on proteins via hydrophilic interactions, effectively solubilizing membrane proteins out of the lipid bilayer (Linke 2009).

Through these mechanisms, Tween 20 permeabilizes and disintegrates cell membranes. It has similar membrane disrupting effects to other nonionic detergents like Triton X-100, but is considered more gentle. This makes Tween 20 well-suited for cell lysis applications.

Using Tween 20 for Cell Lysis and Protein Solubilization

What are some Applications of Tween 20 for Cell Lysis?

Some common applications of Tween 20 for efficient cell lysis include:

Isolation of cytosolic proteins from mammalian cell cultures
Membrane protein extraction from tissues
Lysis of bacterial cells for recombinant protein purification or DNA isolation
Yeast cell lysis for protein analysis
Enveloped virus inactivation by membrane disruption
Cell permeabilization for intracellular staining assays
Plant or algal cell lysis for protein or metabolite extraction

Tween 20 is highly effective at disrupting plasma membranes while leaving organelles and nuclei intact. This selective lysis allows clean isolation of cytosolic fractions. It can also solubilize membrane proteins while avoiding cytosolic protein aggregation that strong ionic detergents can cause.

By optimizing Tween 20 concentrations, high yields of both soluble and membrane-associated proteins can be obtained from lysed cells for downstream analysis.

What Concentration of Tween 20 Solubilizes Membrane Proteins?

The amount of Tween 20 needed to solubilize membrane proteins depends on factors like protein abundance, membrane composition, and the presence of other solubilizing agents. However, concentrations of 0.5-2% Tween 20 are typically used.

Higher concentrations in the 2-5% range can sometimes be needed for effective extraction of highly hydrophobic membrane proteins from lipid-rich membranes (Linke 2009). For example, 3.5% Tween 20 was required to solubilize rhodopsin from rod outer segment mammalian cell membranes (Litman 1982).

Milder 0.5-1% Tween 20 levels suffice for solubilization of moderately hydrophobic membrane proteins, like transporters and receptors, from tissue or cell cultures (Cao et al. 2009; Wu et al. 2019). Addition of salt to disrupt protein-protein interactions or chelators like EDTA also enhances solubilization.

For most membrane protein applications, a starting Tween 20 concentration of 1% is recommended, which can be further optimized as needed. Excessive amounts can sometimes interfere with purification methods. Overall, Tween 20 provides a versatile, nondenaturing option for membrane protein solubilization.

What are Alternatives to Tween 20 for Gentler Cell Lysis?

While excellent for most routine lysis applications, Tween 20 does have some drawbacks. The nonionic detergent can sometimes cause excessive membrane dissolution or protein denaturation at higher concentrations required for more resistant cells.

For more delicate mammalian cells, nonionic alternatives like Triton X-100 or digitonin can lyse cells under milder conditions. The charged detergent sodium dodecyl sulfate (SDS) also lyses mammalian cells but denatures proteins.

Quaternary ammonium detergents like cetyltrimethylammonium bromide (CTAB) work well for plant or algal cells at just 0.5-1% concentrations and maintain protein integrity (Eland et al. 2012; Puri et al. 2012).

For bacterial lysis, lysozyme digestion followed by Triton X-100 solubilization allows gradual, controlled lysis without cellular protease release (Berrada et al. 1996).

Overall, Tween 20 remains a versatile first choice for most cell lysis applications. But exploring alternate detergents can optimize results for finicky cell types or proteins.

Using Tween 20 to Inactivate Enveloped Viruses

Does Tween 20 Inactivate Enveloped Viruses?

Yes, Tween 20 is highly effective at inactivating enveloped viruses like influenza, HIV, herpesviruses, and coronaviruses. At just 0.1-1% concentrations, Tween 20 can disrupt the lipid envelope and membrane proteins of the virus, rendering it non-infectious.

Several studies confirm the potent virucidal effects of Tween 20. For example, 0.1% Tween 20 inactivated over 99.99% of HIV, hepatitis B, and herpes simplex virus particles within 5 minutes (Date et al. 2010). Influenza virus also lost infectivity within 1 minute of Tween 20 treatment (Roy et al. 2020).

Tween 20 is even effective against enveloped viruses like SARS-CoV-2 at ambient temperatures, making it a promising disinfectant (Hulkower et al. 2020). It likely exerts its antiviral effects by dissolving the viral envelope.

What is the Mechanism for Viral Inactivation?

As a detergent, Tween 20 can disrupt the lipid envelopes around viruses, similar to how it damages cell membranes. The viral envelope is essential for entry and fusion with host cells.

Additionally, Tween 20 is thought to irreversibly bind and solubilize the membrane glycoproteins protruding from the viral envelope (Thompson et al. 2017). These proteins mediate attachment and entry into host cells.

By disintegrating the viral envelope and membrane proteins, Tween 20 potently inactivates enveloped virions through biophysical means. The virucidal effects occur rapidly at low concentrations compatible with biological samples.

How is Tween 20 Used for Viral Inactivation?

Some common uses of Tween 20 for viral inactivation include:

Disinfecting surfaces contaminated with enveloped viruses like SARS-CoV-2
Incorporating into virucidal hand sanitizers
Pretreating serum or plasma donations to prevent enveloped virus transmission
Inactivating viral samples for vaccine production or research use
As part of viral transport media to preserve viral RNA/DNA while inactivating infectious particles
Coating ELISA plates to prevent viral retention and signal loss

Only brief Tween 20 exposure is needed for potent virucidal effects. However, Tween 20 does not inactivate non-enveloped viruses like norovirus, poliovirus, or adenovirus lacking the lipid envelope. It provides a simple, effective means to combat transmission of deadly enveloped viruses.

Conclusion

In summary, substantial evidence confirms that Tween 20 is an effective detergent for lysing diverse cell types and solubilizing membrane proteins, in addition to inactivating enveloped viruses. It disrupts membranes through a hydrophobic effect and is commonly utilized at 0.1-1% in cell lysis buffers. Tween 20 concentration should be optimized based on cell sensitivity. More resistant bacteria, yeast, and plant cells tolerate higher levels nearing 1-2% Tween 20 for lysis. For membrane protein studies, 1-3% Tween 20 is typically employed. Lastly, Tween 20 rapidly neutralizes enveloped viruses like HIV and SARS-CoV-2 by just 0.1-0.5% concentrations via membrane damage. While not appropriate for every application, Tween 20 is a workhorse detergent for many biochemical techniques involving cell lysis, protein analysis, and viral inactivation.

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