Protein Standard Multi-Modal Detection Gel Electrophoresis

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Solution Overview

Problem

Current protein gel electrophoresis techniques face challenges in sensitive and specific detection of proteins, particularly in characterizing molecular weight and purity, due to limitations in existing protein standards and detection methods.

Innovation Solution

A protein standard comprising multiple protein sets with unlabeled and labeled proteins, where the labeled proteins have spectrally distinct haloalkylated tryptophan or covalently linked dyes, allowing for multi-modal detection and characterization, including UV-induced fluorescence and visible light detection.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If traditional protein standards are used for gel electrophoresis, then the technique is simple and cost-effective, but the detection sensitivity and specificity are insufficient

Engineering Contradiction:
Improvedetection sensitivityVSAvoidstandard composition complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The protein standard is divided into multiple protein sets, where each set contains proteins with specific molecular weights and detection characteristics. This segmentation allows different protein sets to be detected by different modalities (UV fluorescence, visible light, mass spectrometry) simultaneously, improving overall detection sensitivity without requiring a single complex standard

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The protein standard comprises a composite mixture of labeled and unlabeled proteins with different properties. Labeled proteins contain specific amino acid motifs or covalently attached chemical groups that enable detection by various methods, while unlabeled proteins provide molecular weight references. This composite structure enhances both detection sensitivity and versatility

Inventive Principle:
Principle #40Composite materials

2Adaptability or versatility

If protein standards with multiple labels are used, then multi-modal detection is enabled, but the spectral overlap between labels increases

Engineering Contradiction:
Improvedetection modality versatilityVSAvoidspectral distinction accuracy
Core Design Contradiction:
Adaptability or versatilityVSMeasurement precision

Solution Approach 1:

Different protein sets within the standard are assigned different local detection qualities - some are optimized for UV fluorescence detection, others for visible light absorption, and some for mass spectrometry. This local optimization allows each protein set to be detected by its最适合 modality without spectral interference, maintaining measurement precision while enabling multi-modal detection

Inventive Principle:
Principle #3Local quality

3Measurement precision

If labeled proteins are used to enhance detection, then sensitivity improves, but the cost of the protein standard increases

Engineering Contradiction:
Improvedetection sensitivityVSAvoidstandard production cost
Core Design Contradiction:
Measurement precisionVSEase of manufacture

Solution Approach 1:

Instead of labeling all proteins in the standard, only specific protein sets are labeled with detectable groups. This partial labeling approach provides sufficient detection sensitivity for those specific proteins while keeping the overall standard cost-effective. The unlabeled proteins still serve as molecular weight references

Inventive Principle:
Principle #16Partial or excessive action

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

Enables robust and precise characterization of proteins through multiple detection modalities, improving sensitivity and specificity in protein characterization and detection, even in the presence of reducing agents.

Implementation Method 1

said unlabeled protein comprising a tryptophan residue, and said labeled protein comprising a label that is spectrally distinct from haloalkylated tryptophan

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Implementation Method 2

the label of the labeled protein is an amino acid residue covalently linked to a porphyrin... The label is an amino acid residue covalently linked to a dye

Methodology Applied
Scientific EffectAbsorption spectroscopy: Absorption Spectroscopy

Data Source

PatentEP2976622B1Protein standard
Publication Date: 2019.11.27 BIO RAD LABORATORIES INC
  • EP2976622B1 patent drawingFigure 1
  • EP2976622B1 patent drawingFigure 2A~2C

AI summary

Disclosed herein is a protein standard for gel electrophoresis. The standard may be detected using multiple modalities. These modalities include, for example, observation of color or fluorescence arising from dyes, porphyrins, or haloalkylated tryptophan residues covalently linked to proteins of the standard; or detection of fluorescence or chemiluminescence arising from antibodies or other binding partners bound to proteins of the standard through polypeptide epitopes or affinity tags. The protein standard comprises multiple protein sets, each set corresponding to a different gel band, and proteins within a set may be detectable with one or more modalities.