Outer Membrane Nanovesicle Preparation With Sarkosyl Purification

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Solution Overview

Problem

Existing methods for producing Gram-negative bacterial outer membrane-derived nanovesicles (OMNVs) face challenges in mass production due to complexity and the need for substances like lysozyme, and they contain unwanted intracellular substances that reduce efficiency and cause adverse effects.

Innovation Solution

A method involving the use of Sarkosyl to remove the inner membrane from Gram-negative bacteria, allowing for the extraction of the outer membrane to prepare nanovesicles, which are then purified to enhance yield and purity, and can be loaded with therapeutic or diagnostic substances.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional methods (EDTA, lysozyme, sonication, high-speed centrifugation) are used to produce OMNVs, then outer membrane can be obtained, but the process is complex and difficult resulting in low yields

Engineering Contradiction:

Improveyield of OMNVsVSAvoidcomplexity of purification process

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The invention extracts and removes the inner membrane and intracellular substances from Gram-negative bacteria using Sarkosyl treatment, isolating only the outer membrane to form nanovesicles. This extraction approach simplifies the production process by directly obtaining pure outer membrane vesicles without complex multi-step purification, thereby increasing yield while reducing process complexity

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention changes the chemical parameter by using Sarkosyl (a detergent) instead of the conventional EDTA and lysozyme combination. This parameter change fundamentally alters the membrane disruption mechanism, enabling selective removal of inner membrane and intracellular contents while preserving outer membrane integrity, thus simplifying the overall process and improving productivity

Inventive Principle:

Principle #35Parameter changes

2Manufacturing precision

If conventional purification methods are used, then outer membrane vesicles can be isolated, but unnecessary intracellular substances remain that reduce efficiency and induce adverse effects

Engineering Contradiction:

Improvepurity of OMNVsVSAvoidadverse effects from intracellular substances

Core Design Contradiction:

Manufacturing precisionVSObject-affected harmful factors

Solution Approach 1:

The invention converts the harmful effect of Sarkosyl (a detergent that can disrupt membranes) into a beneficial selective removal tool. By controlling the treatment conditions, Sarkosyl selectively removes inner membrane and intracellular substances while preserving outer membrane vesicles, thereby purifying the product and eliminating harmful intracellular contaminants that would otherwise cause adverse effects

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

Solution Approach 2:

Sarkosyl acts as an intermediary agent that mediates the separation between outer membrane vesicles and intracellular substances. The detergent facilitates selective removal of unwanted intracellular components while leaving the outer membrane intact, achieving high purity OMNVs without requiring multiple complex purification steps

Inventive Principle:

Principle #24Intermediary (Mediator)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The method achieves a higher yield and purity of OMNVs, reducing adverse effects and enhancing therapeutic efficacy by delivering targeted substances to cells or tissues.

Implementation Method 1

treating Gram-negative bacteria with Sarkosyl removes the inner membrane

Methodology Applied

Scientific EffectDetergent action: Surfactant

Implementation Method 2

manufacturing through sonication

Methodology Applied

Scientific EffectSonication: Ultrasonic Vibration

Implementation Method 3

separating the outer and inner membranes using high-speed centrifugation

Methodology Applied

Scientific EffectCentrifugal separation: Centrifugal Force

Data Source

PatentUS20260014086A1Method for constructing gram-negative bacterial outer membrane-derived nanovesicles and use thereof

Publication Date: 2026.01.15 POSTECH ACADEMY INDUSTRY FOUNDATION

AI summary

The present invention relates to a method for preparing Gram-negative bacterial outer membrane-derived nanovesicles (OMNVs) and uses thereof. Additionally, the present invention provides a method for treating or diagnosing cancer using OMNVs. Furthermore, the present invention provides a method for delivering therapeutic or diagnostic substances to target cells or tissues by loading them into OMNVs. Also, the present invention provides a method for preventing and treating various diseases, including cancer and bacterial/viral infections, by co-administering OMNVs with antigens or administering OMNVs expressing antigens.