Cellular Component Purification via Electric Field pH Gradients

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Solution Overview

Problem

Current isoelectric focusing technologies face limitations in achieving optimal protein, peptide, nucleic acid, organelle, and cell fractionation due to fixed pH ranges and chemical contamination from ampholytes, which hinder sample separation and downstream analysis.

Innovation Solution

The use of proton and hydroxide injectors with bipolar membranes to generate pH gradients and separate cellular components based on their isoelectric points, allowing for precise positioning and collection of target molecules without chemical contamination, using a system with controlled pH gradients and electrical fields.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If chemical ampholytes are used to establish pH gradients for sample fractionation, then separation of cellular components is achieved, but sample contamination with chemicals occurs and downstream analysis is interfered with

Engineering Contradiction:

Improvefractionation qualityVSAvoidchemical contamination

Core Design Contradiction:

Manufacturing precisionVSObject-affected harmful factors

Solution Approach 1:

The patent extracts and removes the harmful chemical ampholytes from the fractionation process. Instead of using chemical pH gradients, the invention employs electric fields to directly control pH through proton and hydroxide injectors, thereby achieving separation without chemical contamination that interferes with downstream analysis.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent replaces the chemical mechanism (ampholyte-based pH gradients) with an electrical mechanism (electric field-controlled proton and hydroxide injection). This substitution eliminates chemical contamination while maintaining the ability to establish precise pH gradients for effective fractionation of cellular components.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

2Device complexity

If fixed or limited pH range is used for sample separation, then fractionation process is simplified, but optimal separation of various samples is not achieved

Engineering Contradiction:

Improvefractionation process complexityVSAvoidseparation quality

Core Design Contradiction:

Device complexityVSManufacturing precision

Solution Approach 1:

The patent implements dynamic pH gradient adjustment through programmable proton and hydroxide injectors controlled by a processor. The system can adaptively modify pH ranges and gradients based on specific sample requirements, enabling optimal separation of diverse cellular components while maintaining manageable system complexity through automated control.

Inventive Principle:

Principle #15Dynamics

Solution Approach 2:

The patent enables flexible modification of pH gradient parameters (range, slope, distribution) through electric field control. By changing electrical parameters injected into the medium, the system can optimize separation conditions for different sample types without requiring complex physical reconfiguration, thus improving separation quality while keeping the process relatively simple.

Inventive Principle:

Principle #35Parameter changes

3Manufacturing precision

If chemical ampholytes are used to create pH gradients, then sample fractionation is enabled, but downstream analysis is interfered with by chemical presence

Engineering Contradiction:

Improvefractionation capabilityVSAvoiddownstream analysis accuracy

Core Design Contradiction:

Manufacturing precisionVSReliability

Solution Approach 1:

The patent extracts and eliminates chemical ampholytes from the fractionation process, using electric field-controlled proton and hydroxide injection instead. This removal of chemical contaminants ensures that downstream analysis of separated cellular components is not interfered with, thereby improving analysis reliability while maintaining effective fractionation capability.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent introduces electric fields as an intermediary mechanism to establish pH gradients without requiring chemical ampholytes. The electrical field acts as a mediator that enables fractionation while leaving no residual chemical contaminants, thus ensuring both fractionation capability and downstream analysis reliability.

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

This method enables efficient and contamination-free separation and purification of cellular components, such as proteins, nucleic acids, and organelles, by positioning them based on their isoelectric points, facilitating downstream analysis without the interference of chemical ampholytes.

Implementation Method 1

generating a pH gradient or pH step in the chamber with one or more proton injector(s) and/or hydroxide injector(s), such that at least two components from the cells in different positions are positioned in the chamber based on the isoelectric point (pI) of the components

Methodology Applied

Scientific EffectIsoelectric focusing: Isoelectric Focusing