Electrophoresis Techniques for Plant Protein Extraction
Plant protein extraction presents significant technical hurdles due to the complex cellular matrices and diverse protein structures involved. Current methods achieve protein yields of 40-60% with varying degrees of functionality, while maintaining protein integrity remains challenging when processing volumes exceed 100L. The presence of cell wall components, phenolic compounds, and other interfering substances often compromises extraction efficiency and protein quality.
The fundamental challenge lies in balancing extraction efficiency with protein functionality while managing the trade-offs between processing intensity and protein denaturation.
This page brings together solutions from recent research—including membrane separation coupled with physical field modification, triple-electrode electrophoresis configurations, enzymatic lysis combined with chemical treatments, and novel two-dimensional separation techniques. These and other approaches focus on achieving higher protein yields while preserving functional properties essential for commercial applications.
1. Method for Preparing Plant Proteins via Membrane Separation and Physical Field Modification
OIL CROPS RES INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES, 2024
A novel method for preparing highly functional plant proteins through membrane separation coupled physical field modification. The process involves membrane separation to obtain protein concentrate, followed by controlled alkaline treatment to enhance protein structure, and subsequent physical field modification using ultrasound and infrared to further improve protein properties. This method enables the preparation of proteins with enhanced solubility, foaming, and emulsifying capabilities, while maintaining their nutritional value and structural integrity.
2. Triple-Electrode Electrophoresis Device with Annular Electrode Configuration for Biomolecule Enrichment and Collection
Jiangmen Canming Biotechnology Co., Ltd., 2024
A triple-electrode electrophoresis device for enhanced extraction of biomolecules from large-volume samples. The device features a tank body with three annular electrodes: a central enrichment electrode, a ring enrichment electrode, and a ring collection electrode. The central and ring electrodes have the same polarity, while the ring electrode has the opposite polarity. This configuration creates a unique enrichment zone where biomolecules concentrate, while the ring collection electrode captures the enriched molecules. The device's tank body contains a collection system for the enriched biomolecules, allowing for efficient sample processing in medium to large-volume samples.
3. Method for Protein Extraction via Chloroplast Membrane Disruption and Divalent Ion Precipitation
STICHTING WAGENINGEN RESEARCH, 2022
A method for extracting functional proteins from plant materials that retains their nutritional and functional properties. The method involves disrupting chloroplast membranes in plant juice, followed by selective precipitation of chloroplast membranes using a divalent ion. This selective precipitation process enables the removal of chloroplast membranes while preserving the soluble protein, resulting in a protein concentrate with minimal impurities. The method can be performed on a large scale and is particularly effective for obtaining functional proteins from plant materials like sugar beets.
4. Process for Producing High-Purity Protein Extracts from Plant Materials Using Enzymatic Lysis and Chemical Treatment
PLANTIBLE FOODS INC, 2021
A process for producing high-purity protein extracts from plant materials, enabling the creation of stable and functional protein preparations for various food applications. The method employs a combination of enzymatic lysis and chemical treatment to generate a high-quality protein solution, followed by selective purification steps to achieve a purity of at least 80%. The process leverages plant-specific enzymatic activity to break down the plant material, followed by chemical treatment to remove contaminants. The purified protein is then further purified through selective adsorption, filtration, and ultrafiltration, resulting in a protein preparation with minimal impurities and excellent functional properties.
5. Electrophoretic Analyzer with Integrated Buffer System and Membrane-Based Transfer for Direct Plate Separation
UNIVERSITY OF ULSAN FOUNDATION FOR INDUSTRY COOPERATION, 2019
An electrophoretic analyzer for analyzing organic materials through a novel approach that eliminates the need for gel transfer steps. The device integrates a buffer system into the electrophoresis chamber, allowing the separation of organic materials directly on the plate. The buffer system maintains optimal conditions for protein separation while preventing gel damage and maintaining membrane integrity. The device features a membrane-based transfer system, precision voltage control, and a circulation system to maintain optimal conditions throughout the analysis process. This eliminates the need for gel handling and transfer steps, reducing analysis time and complexity.
6. Electrophoresis Tank with Three-Buffer Electrode Configuration and Integrated Column Holder for Protein Separation
RESEARCH CENTER FOR ECO-ENVIRONMENTAL SCIENCES CHINESE ACADEMY OF SCIENCES, 中国科学院生态环境研究中心, 2019
A novel electrophoresis tank and column system for protein separation and detection that enables high-performance, high-throughput analysis of complex biological samples. The tank features a unique configuration with three buffer tanks positioned on the electrodes, allowing precise control of pH and ionic strength conditions. The tank is mounted on a column holder, which enables precise positioning of the column. The system incorporates a dedicated cooling system and elution solution delivery system, enabling rapid and efficient sample processing. The tank's design enables the simultaneous use of ICP-MS detection, making it an ideal platform for comprehensive protein analysis.
7. Protein Separation Method Utilizing Two-Dimensional Polyacrylamide Gel Electrophoresis for Broussonetia Papyrifera Leaves
INSTITUTE OF BOTANY THE CHINESE ACADEMY OF SCIENCES, Institute of Botany, Chinese Academy of Sciences, 2019
Separating protein from Broussonetia papyrifera leaves using protein two-dimensional polyacrylamide gel electrophoresis. The method employs 2-DE to separate and analyze proteins from Broussonetia papyrifera leaves, providing a reliable and efficient method for studying plant proteome. The technique enables precise protein separation and identification, particularly challenging in plant-based protein separation applications.
8. Planar Electrophoresis Device with Integrated Ion-Exchange and Electrostatic Interaction Mechanisms for Protein Separation
Hualan Biological Engineering Chongqing Co., Ltd., Hualan Biological Engineering Co., Ltd., HUALAN BIOENGINEERING CHONGQING CO LTD, 2019
A device for efficient batch separation of protein mixtures through a single-step, planar electrophoresis process. The device employs a novel combination of ion-exchange and electrostatic interactions to achieve simultaneous separation of multiple protein components. The separation process is enabled by a unique combination of ion-exchange and electrostatic interactions between protein molecules and the device's surface, allowing for rapid and efficient separation of complex protein mixtures.
9. Multi-Row Horizontal Continuous Electrophoresis System with Parallel Polyacrylamide Gel Rows
Peking University, 2018
A multi-row high-throughput horizontal continuous electrophoresis polyacrylamide gel system enables simultaneous detection of hundreds of samples in a conventional gel size range. The system employs a single gel plate with multiple parallel rows of polyacrylamide gels, each containing a uniform separation gel and an electrophoresis buffer. The gel buffer composition is optimized for high-throughput protein analysis while maintaining consistent migration characteristics across rows. The system achieves parallel detection of multiple samples simultaneously, enabling applications like protein quantification, parallel comparison of multiple samples, and large-scale screening.
10. Device for Single-Step Continuous Electrophoresis with Combined Isoelectric Point and Molecular Weight Separation
HUALAN BIOLOGICAL ENGINEERING INC, 2018
A protein separation and purification method that enables efficient and precise batch processing of complex protein mixtures through a single-step, continuous electrophoresis process. The method employs a device that combines multiple separation steps in a single run, allowing for simultaneous separation of multiple protein components based on their isoelectric point and molecular weight. This approach eliminates the need for multiple purification steps, reducing overall processing time and improving purity compared to traditional sequential purification methods.
11. Two-Dimensional Electrophoresis System with Solvent Extraction for Salt Ion Removal in Plant Extracts
HUBEI HOUYOU INTELLECTUAL PROPERTY SERVICE CO LTD, 2018
A two-dimensional electrophoresis system for Suaeda salsa that eliminates salt accumulation during sample preparation. The system employs a novel solvent extraction method that selectively removes salt ions from plant extracts without requiring specialized equipment. This approach enables efficient protein extraction from high-salt plant samples like Suaeda salsa, enabling reliable proteomic analysis.
12. Protein Extraction Method for Lily Tissues Using Optimized Acetone Solvent Protocol
FLOWER RESEARCH INSTITUTE YAAS, Yunnan Academy of Agricultural Sciences Flower Research Institute, FLOWER RESEARCH INSTITUTE, YAAS, 2018
Optimizing protein extraction and two-dimensional electrophoresis for lily plants to produce high-quality protein profiles. The method employs a novel extraction protocol that improves protein yield and quality by optimizing the use of acetone, a common extraction solvent for plant proteins. This protocol overcomes common challenges in protein extraction, enabling the isolation of high-quality total protein from lily tissues. The extraction protocol is then followed by two-dimensional electrophoresis, where the extracted proteins are separated based on their isoelectric point and molecular weight. The resulting differential protein map provides detailed insights into the protein composition of lily tissues under stress conditions, enabling researchers to better understand the molecular mechanisms underlying disease resistance in lily plants.
13. Protein Classification Device with Rotating Drum and Synchronized Disc Separation System
TIANJIN THIRD CENTER HOSPITAL, Tianjin Third Central Hospital, 2018
A device for precise protein classification and processing that enables efficient separation of plant proteins through a compact, single-stage system. The device features a rotating drum with multiple through holes, a motorized drum rotation system, and a series of rotating discs. The system uses a rotating drum to pass through the discs, with the discs rotating in synchronization to separate proteins based on size and composition. The separation process is optimized through precise control of the drum rotation speed and the positioning of the discs. The system includes a crushing press device, filtering device, and chromatography column, all integrated into a single frame that is mounted on a fixed frame.
14. Method for Extracting Soluble Leaf Proteins Using pH-Stabilizing Buffer Solution
UNIVERSITY OF MARYLAND COLLEGE PARK, 2018
A novel method for extracting soluble leaf proteins from plant leaves for food and industrial applications. The process employs a buffer solution that maintains protein solubility throughout the extraction process, allowing the removal of non-protein components without protein denaturation. The buffer solution maintains optimal pH conditions for protein solubility while preventing protein aggregation. The method is scalable and can be adapted to various plant species, including tobacco, alfalfa, and spinach.
15. Method for Protein Separation and Purification Using Two-Dimensional Gel Electrophoresis with Non-Reducing Isoelectric Focusing and Low SDS Concentration
ZIBO ARRAY BRIDGE BIOLOGICAL TECHNOLOGY CO LTD, Zibo Yunqiao Biotechnology Co., Ltd., 2017
Method for separating and purifying proteins from complex mixtures like cell extracts using two-dimensional gel electrophoresis (2-DE) without losing protein activity. The method involves running the first-dimension isoelectric focusing (IEF) gel without reducing agents to preserve disulfide bonds. For the second-dimension SDS-PAGE, the SDS concentration is reduced to 0.1%. This allows efficient separation of proteins while maintaining their enzymatic activity. After 2-DE, proteins are recovered from the gel using a specially designed protein elution plate with many small wells. This allows isolating individual proteins from the complex mixture. The plate has molecular semipermeable membranes that permit small molecules to pass through while retaining the proteins. This provides a simple, high-resolution method for isolating and purifying proteins from complex samples using 2-
16. Two-Dimensional Gel Electrophoresis with Gel Eluted Liquid Fraction Entrapment and Isoelectric Focusing
UNIV SAINS MALAYSIA, 2017
A two-dimensional gel electrophoresis method for separating proteins from complex biological samples, particularly for analysis of periplasmic proteins in Sargassum. The method employs gel eluted liquid fraction entrapment electrophoresis in the first dimension followed by isoelectric focusing in the second dimension. The separation is achieved through a novel approach where fractions are trapped in subsequent liquid phases, allowing precise resolution of proteins based on their charge. The method enables the separation of proteins differing in charge, including those with single charge changes, and is particularly suitable for analyzing proteins from complex biological sources like ribosomal assembly, plant extracts, and highly insoluble proteins like periplasmic proteins.
17. Gradient Gel Electrophoresis System for Differential Protein Migration by Molecular Weight
NANCHANG UNIVERSITY, Nanchang University, 2017
A novel gradient electrophoresis method for separating and identifying native protein molecules through differential migration based on molecular weight. The method employs a gradient gel system with a unique composition that enables the separation of proteins with varying molecular weights. The gradient composition is optimized to maintain protein integrity during migration while selectively allowing proteins of different molecular weights to migrate at distinct rates. This approach enables the precise identification of proteins with distinct molecular weights, previously challenging to separate using conventional electrophoresis techniques.
18. Precipitation Method for Selective RuBisco Removal from Plant Materials Using PEG4000 Solution
Institute of Botany, the Chinese Academy of Sciences, 2016
A method for removing leaf protein RuBisco from plant materials through precipitation using a 300-500g/L PEG4000 solution. The method enables efficient protein removal while preserving the basic protein structure and separating low-abundance proteins. The PEG precipitation process selectively precipitates RuBisco protein, eliminating its masking effects on other proteins during two-dimensional gel electrophoresis. This approach optimizes protein extraction from plant materials like mulberry leaves for proteomic analysis.
19. Protein Separation Platform Utilizing Combined Electrostatic and Hydrophobic Interaction Mechanism
ZICHAO DANG, Dang Zichao, 2015
Separation and purification of proteins through a novel, high-performance method that enables efficient and precise protein separation and purification in complex biological systems. The method employs a novel separation platform that utilizes a combination of electrostatic and hydrophobic interactions to selectively separate proteins from complex biological matrices. The platform achieves high resolution separation and purification of proteins while maintaining their native structure and function, overcoming conventional separation limitations.
20. Detergent-Free Protein Purification Method Using Denaturing Gel Electrophoresis and Solubilization-Precipitation Technique
SHANGLUO UNIVERSITY, 2015
A novel method for protein purification that addresses the challenges of protein separation and purification through a combination of denaturing gel electrophoresis and detergent-free purification. The method employs a unique detergent-free approach that preserves protein structure and function while achieving high-resolution separation of proteins through gel electrophoresis. The purification step utilizes a combination of detergent-free solubilization and detergent-free precipitation to separate proteins from non-protein contaminants. This approach enables the purification of thousands of protein variants from complex biological samples, overcoming the limitations of traditional gel-based purification methods.
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