Multimeric Protein Complex for Porphyromonas gingivalis Infection

Solution Overview

Problem

Current methods for addressing periodontal diseases associated with Porphyromonas gingivalis are limited in effectively targeting the bacterium's virulence factors, particularly the trypsin-like proteases, which contribute to disease progression and severity.

Innovation Solution

A purified multimeric protein complex comprising domains from RgpA, Kgp, and HagA, with a molecular weight greater than 300 kDa, is extracted from P. gingivalis cells using detergent extraction and further purified through ion exchange or ultrafiltration, forming a high molecular weight proteinase-adhesin complex for use in eliciting an immune response and treating infections.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If traditional methods are used to address periodontal diseases, then the approach is simple and easy to implement, but the effectiveness in targeting virulence factors is insufficient

Engineering Contradiction:

Improveeffectiveness in targeting virulence factorsVSAvoidcomplexity of treatment approach

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent extracts and isolates specific virulence factors (trypsin-like proteases RgpA and Kgp, and hemagglutinin HagA) from the complex bacterial cell surface proteins of P. gingivalis. These extracted proteins are then used as vaccine antigens to induce targeted immune responses against the virulence factors, thereby resolving the contradiction by focusing the immune response on specific harmful proteins rather than the entire bacterium.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent segments the bacterial cell surface protein complex into distinct functional components: RgpA (arginine-specific trypsin-like protease), Kgp (lysine-specific trypsin-like protease), and HagA (hemagglutinin). This segmentation allows the immune system to target and neutralize each virulence factor independently, improving the effectiveness of the treatment while maintaining manageable complexity through modular vaccine design.

Inventive Principle:

Principle #1Segmentation

2Reliability

If a multimeric protein complex is extracted and purified, then the immune response is significantly improved, but the purification process becomes more complex

Engineering Contradiction:

Improveimmune response effectivenessVSAvoidease of purification process

Core Design Contradiction:

ReliabilityVSEase of manufacture

Solution Approach 1:

The patent merges multiple protein components (RgpA, Kgp, and HagA) into a single multimeric complex structure on the bacterial cell surface. This merging allows the extraction and purification processes to handle multiple virulence factors simultaneously as a unified complex, simplifying the manufacturing process while maintaining the immunogenicity and effectiveness of the vaccine against all components.

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The patent utilizes composite protein structures where RgpA, Kgp, and HagA are assembled into a multimeric complex on the P. gingivalis cell surface. This composite structure provides both structural stability and immunogenicity, enabling effective vaccine formulation while the composite nature allows for coordinated purification through techniques that maintain the complex's intact structure.

Inventive Principle:

Principle #40Composite materials

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 complex induces a significant immune response and provides protection against P. gingivalis infections, reducing disease severity and preventing periodontal bone loss in animal models, offering a more effective approach than traditional methods.

Implementation Method 1

The complex can be extracted from P. gingivalis cells by detergent extraction or ultrasonication followed by ultrafiltration/diafiltration or anion exchange and Lys-sepharose or Arg-sepharose chromatography.

Methodology Applied

Scientific EffectDetergent extraction: Surfactant

Implementation Method 2

The complex can be extracted from P. gingivalis cells by detergent extraction or ultrasonication followed by ultrafiltration/diafiltration or anion exchange and Lys-sepharose or Arg-sepharose chromatography.

Methodology Applied

Scientific EffectIon exchange: Ion Exchange

Implementation Method 3

The complex can be extracted from P. gingivalis cells by detergent extraction or ultrasonication followed by ultrafiltration/diafiltration or anion exchange and Lys-sepharose or Arg-sepharose chromatography.

Methodology Applied

Scientific EffectUltrafiltration: Filter (physical)

Implementation Method 4

The complex can be extracted from P. gingivalis cells by detergent extraction or ultrasonication followed by ultrafiltration/diafiltration or anion exchange and Lys-sepharose or Arg-sepharose chromatography.

Methodology Applied

Scientific EffectUltrasonication: Ultrasonic Vibration

Data Source

PatentUS8765144B2Antigenic complex for the diagnosis and treatment of <i>Porphyromonas gingivalis </i>infection

Publication Date: 2014.07.01 UNIVERSITY OF MELBOURNE

AI summary

The present invention provides a purified multimeric complex from P. gingivalis. The complex comprises at least one domain from each of RgpA, Kgp and HagA, and has a molecular weight greater than about 300 kDa.