Assembly of Cell Mimicking Supported and Suspended Lipid Bilayer Models for the Study of Molecular Interactions

Christina M. Bailey-Hytholt; Veronica LaMastro; Anita Shukla

doi:10.3791/62599

JoVE Journal
Bioengineering

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Bioengineering

Assembly of Cell Mimicking Supported and Suspended Lipid Bilayer Models for the Study of Molecular Interactions

分子相互作用研究のための細胞模倣支援及び懸濁脂質二重層モデルの組み立て

Full Text

4,175 Views

12:18 min

August 3, 2021

DOI: 10.3791/62599-v

Christina M. Bailey-Hytholt¹, Veronica LaMastro², Anita Shukla²

¹Department of Chemical Engineering,Worcester Polytechnic Institute, ²Center for Biomedical Engineering, School of Engineering,Brown University

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol describes the formation of cell mimicking uni-lipid and multi-lipid vesicles, supported lipid bilayers, and suspended lipid bilayers. These in vitro models can be adapted to incorporate a variety of lipid types and can be used to investigate various molecule and macromolecule interactions.

Key Study Components

Area of Science

Neuroscience
Biophysics
Biochemistry

Background

Lipid bilayers are essential for studying membrane interactions.
These models can simulate biological membranes for various applications.
Understanding membrane interactions is crucial for drug development.
Different lipid compositions can affect membrane properties.

Purpose of Study

To create versatile lipid bilayer models for research.
To investigate interactions with pharmaceutical agents and toxicants.
To assess compound permeability and absorption in membranes.

Methods Used

Preparation of lipid stock solutions.
Removal of chloroform using nitrogen gas.
Vacuum drying of lipid films.
Rehydration with Tris sodium chloride buffer.

Main Results

Successful formation of lipid vesicles with controlled concentrations.
Demonstrated methods for assessing membrane interactions.
Provided insights into compound embedment within membranes.
Highlighted the adaptability of the protocol for various lipids.

Conclusions

The protocol offers a reliable method for lipid bilayer construction.
It can be used to study a wide range of membrane interactions.
These models are valuable for both basic and applied research.

Frequently Asked Questions

What are lipid bilayers used for?

Lipid bilayers are used to study membrane interactions and properties in various biological and pharmaceutical contexts.

How are the lipid vesicles prepared?

Lipid vesicles are prepared by dissolving lipids, removing solvents, and rehydrating with buffer solutions.

Can different lipids be used in this protocol?

Yes, the protocol can be adapted to incorporate various lipid types.

What is the significance of studying membrane interactions?

Studying membrane interactions is crucial for understanding drug delivery and toxicity mechanisms.

How does the protocol ensure complete removal of chloroform?

Chloroform is removed by applying nitrogen gas and vacuum drying for several hours.

What is the final concentration of vesicles in this protocol?

The final vesicle concentration is 2.5 milligrams per milliliter.

このプロトコルは、ユニ脂質および多脂質小胞、支持脂質二重層、および懸濁された脂質二重層を模倣する細胞の形成を記述する。これらの in vitro モデルは、様々な脂質タイプを組み込むために適応することができ、様々な分子および高分子相互作用を調査するために使用することができる。

このプロトコルに記載されている脂質二重層膜の構築は、医薬品、環境毒性物質、さらには生物学的化合物との膜相互作用に関する重要な情報を得るために使用することができる。これらの技術は、様々な液体の複雑さの膜を製造するために使用され、膜内の化合物の透過性、吸収、および埋め込みを評価するために使用することができます。まず、脂質ストック溶液の適切な量をクリーンガラスのvileに加え、水分補給後に1ミリリットル当たり2.5ミリグラムの最終小胞濃度を達成します。

窒素ガスの流れを用いて、脂質溶液からクロロホルムを除去する。クロロホルムを完全に除去するために、乾燥した脂質膜を真空に少なくとも4時間接続します。乾燥した脂質膜をトリスナトリウムクロリドバッファーの必要量で水分補給し、1ミリリットル当たり2.5ミリグラムの最終小胞濃度を得るために、約15〜30秒間渦を生じさせる。

View the full transcript and gain access to thousands of scientific videos