Polybrene Lentivirus

BioInnovatise Viral Vector Team

Updated July 15, 2024

You may have heard of polybrene or likely have it listed as a necessary material on your lentivirus packaging protocol. But do you know all the reasons why polybrene is important during the lentivirus packaging process to produce lentiviral cells?

Our viral vector team has assembled the below list of resources related to polybrene usage in the lentivirus packaging process.

Polybrene Usage

As mentioned above, polybrene, found in many lentivirus packaging protocols, is a polymer used to increase transduction efficiency.

In the context of lentivirus packaging and transduction, Polybrene serves several functions:

Facilitating Viral Entry: Polybrene neutralizes the charge repulsion between the negatively charged cell surface and the negatively charged viral particles. This allows the lentivirus to come into closer contact with the target cells, enhancing the efficiency of viral entry.

Increasing Transduction Efficiency: By increasing the attraction between the virus and the cell membrane, polybrene increases the likelihood that the virus will successfully infect the target cells.

Stabilizing Viral Particles: Polybrene can also help to stabilize viral particles in solution, potentially increasing their longevity and effectiveness during the transduction process.

Note: Polybrene is not used during lentivirus transfection process. Transduction and transfection are very different steps in the lentivirus packaging process. Learn more about lentivirus transfection.

The above diagram illustrates the lentivirus packaging process at BioInnovatise. Polybrene is used during the packaging stage #2 in the above diagram.

What If I Add Too Much Polybrene?

The tolerance of lentivirus to variations in polybrene concentration can significantly affect the efficiency of transduction as well as cell viability.

Here’s what you might observe with too much polybrene:

Cytotoxicity: High concentrations of Polybrene can be toxic to cells, leading to reduced cell viability. This can vary depending on the cell type, but excessive polybrene can cause cell membrane damage and apoptosis.
Decreased Transduction Efficiency: Although polybrene enhances viral transduction, too high a concentration can have the opposite effect, potentially due to cytotoxic effects or the aggregation of viral particles, which makes them less effective.

Here’s what you might observe with too little polybrene:

Reduced Transduction Efficiency: Without sufficient polybrene, the charge repulsion between the virus and the cell surface remains high, resulting in lower rates of viral entry and transduction.
Variable Results: Insufficient polybrene can lead to inconsistent transduction efficiency, especially in cell types that are more resistant to viral entry.

Overall Factors Affecting Polybrene Lentivirus Optimization:

Cell Type:

Different cell lines have varying sensitivities to polybrene. Learn more about the various cell lines used for lentivirus packaging.
Primary cells or stem cells might be more sensitive and require lower concentrations.

Viral Construct:

Constructs with different promoters or genes might affect cell viability and transduction efficiency. If you are interested in optimizing your plasmid DNA construct for a higher transduction efficiency, learn about our molecular cloning services.
Constructs designed for high expression might lead to more cytotoxicity, affecting the optimal polybrene concentration.

Multiplicity of Infection (MOI):

High MOI might require lower polybrene to avoid excessive cytotoxicity.
Low MOI might benefit from higher polybrene to enhance transduction efficiency.

What If I Don't Want To Use Polybrene?

Polybrene is commonly used to enhance lentiviral transduction efficiency, but it is not the only option. Other methods and substances can also be used to improve lentiviral transduction such as DEAE-Dextran, protamine sulfate, viral transduction enhancers (e.g. LentiBOOST™, ViraDuctin™, and Lenti-X™ Accelerator), spinoculation, or electroporation. If you are interested in using one of these substitutes to produce your lentivirus packaging cells, let us know when requesting a production.

Let’s get started! Our team is incredibly excited to bring your lentivirus research to life. Learn about our quick turnaround lentivirus packaging services.

If you have any questions about polybrene or how to improve your packaging protocol, contact our team.

Want to learn more about the latest in lentivirus based research? Our colleagues at ScienceDirect and Genetic Engineering & Biotechnology News continuously collect and publish the latest information on lentivirus-based research.

Polybrene Lentivirus

Polybrene Usage

What If I Add Too Much Polybrene?

Overall Factors Affecting Polybrene Lentivirus Optimization:

What If I Don't Want To Use Polybrene?

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