Functionalized Silica Protein Removal via Polyacid Protection

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Solution Overview

Problem

Current methods for removing proteins from reaction mixtures in molecular biology research are either labor-intensive, hazardous, or inefficient, often resulting in low nucleic acid yields and residual enzyme activity that interferes with downstream applications.

Innovation Solution

A method using a composition comprising a polyacid and a functionalized silica solid phase with specific substituent groups that selectively binds and irreversibly adsorbs proteins, preventing nucleic acid binding and ensuring high nucleic acid yields without introducing hazardous materials.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If liquid partitioning method using organic solvents is used for protein removal, then protein removal efficiency is improved, but safety hazards and environmental hazards increase

Engineering Contradiction:
Improveprotein removal efficiencyVSAvoidsafety hazards and environmental hazards
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention changes the chemical parameters of the solid phase surface by introducing specific functional groups (carboxyl, sulfonate, phosphate) that provide selective protein binding capability without requiring hazardous organic solvents. This parameter change enables protein removal through controlled chemical interactions rather than organic phase partitioning.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The invention uses composite materials consisting of silica or polymer beads combined with specific functional groups on the surface. This composite structure provides both the mechanical support of the bead and the selective binding capability of the functional groups, achieving protein removal without hazardous chemicals.

Inventive Principle:
Principle #40Composite materials

2Ease of operation

If silica-based adsorption method is used for protein removal, then convenience of use is improved, but selectivity decreases requiring column technology

Engineering Contradiction:
Improveconvenience of useVSAvoidselectivity
Core Design Contradiction:
Ease of operationVSAdaptability or versatility

Solution Approach 1:

The invention applies local quality by introducing specific functional groups at the surface of solid beads that provide localized protein binding sites. This creates regions of high selectivity on the bead surface while maintaining the overall simplicity of the solid phase system, enabling selective protein removal without complex column technology.

Inventive Principle:
Principle #3Local quality

3Ease of manufacture

If heat inactivation method is used for enzyme removal, then simplicity is improved, but residual enzyme activity and side effects increase

Engineering Contradiction:
ImprovesimplicityVSAvoidcomplete inactivation and absence of side effects
Core Design Contradiction:
Ease of manufactureVSReliability

Solution Approach 1:

The invention introduces a mediator substance - the functionalized solid phase - that facilitates enzyme inactivation and removal through specific chemical interactions. This intermediary provides a controlled mechanism for protein removal that is more reliable than heat inactivation while maintaining simplicity, as the functional groups on the solid phase selectively bind and remove enzymes without causing widespread side effects.

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 effectively removes proteins, maintaining high nucleic acid yields and preventing residual enzyme activity, making it safer, more efficient, and compatible with downstream processes.

Implementation Method 1

a solid phase having a surface which comprises a functionalized silica capable of binding protein

Methodology Applied
Scientific EffectAdsorption: Adsorption

Implementation Method 2

selectively binds and irreversibly adsorbs proteins

Methodology Applied
Scientific EffectIrreversible adsorption: Adsorption

Implementation Method 3

a composition comprising (i) a polyacid comprising a polycarboxylic acid, a polyphosphonic acid, or a polycarboxylate or polyphosphonate salt

Methodology Applied
Scientific EffectChemical interaction:

Data Source

PatentEP3118314B1Protein removal agent
Publication Date: 2018.02.28 THERMO FISHER SCI BALTICS UAB
  • EP3118314B1 patent drawingFigure 1~2
  • EP3118314B1 patent drawingFigure 3~4
  • EP3118314B1 patent drawingFigure 5

AI summary

A composition for the removal of protein contaminants from a solution containing target nucleic acid, which composition comprises: (i) a polyacid, which comprises a polycarboxylic acid, a polyphosphonic acid or a polycarboxylate or polyphosphonate salt; and (ii) a solid phase having a surface which comprises a functionalised silica, wherein the solid phase comprises silica, wherein the solid phase optionally comprises particles, wherein the particles are optionally approximately spherical and have a diameter in the range of from 3 to 15 µm, and wherein the particles optionally have pores with a diameter of from 10 to 100 nm.