Novel Coronavirus N Protein Reference Material Certification

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Solution Overview

Problem

The accuracy and reliability of COVID-19 diagnosis depend on the availability of a reliable reference material for the novel coronavirus nucleocapsid protein (nCOV N protein), which is currently lacking, affecting the consistency and comparability of test results.

Innovation Solution

A method for preparing and certifying a novel coronavirus nucleocapsid protein reference material involves purification, dilution, uniformity testing, and characterization using advanced techniques like isoelectric focusing, molecular sieve methods, and mass spectrometry to ensure high purity and stability, enabling accurate and consistent measurement.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If a commercially available nCOV N protein biochemical reagent with 90-95% purity is used directly, then the preparation process is simple and fast, but the purity and reliability of the reference material are insufficient

Engineering Contradiction:
Improvereliability of reference materialVSAvoidcomplexity of purification process
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The purification process is divided into multiple sequential steps including isoelectric focusing electrophoresis and molecular sieve chromatography. Each step targets specific impurities and progressively increases purity from 90-95% to above 98.5%, resolving the contradiction by breaking down the complex purification into manageable segments that collectively achieve high reliability

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The method performs preliminary purification and concentration steps before final reference material preparation. By pre-treating the commercially available reagent through isoelectric focusing and molecular sieve methods, the system establishes a high-purity base material that ensures subsequent reference material reliability without requiring complex adjustments later

Inventive Principle:
Principle #10Preliminary action

2Manufacturing precision

If advanced purification methods like isoelectric focusing and molecular sieve are applied, then the purity reaches 98.5% or above, but the preparation time and process complexity increase

Engineering Contradiction:
Improvepurity of nCOV N proteinVSAvoidpreparation time
Core Design Contradiction:
Manufacturing precisionVSLoss of time

Solution Approach 1:

The purification process employs continuous flow methods in both isoelectric focusing electrophoresis and molecular sieve chromatography. The system maintains continuous operation through optimized buffer systems and sequential processing, achieving above 98.5% purity while minimizing idle time between steps, thus reducing overall preparation time despite the advanced methods used

Inventive Principle:
Principle #20Continuity of useful action

Solution Approach 2:

The method optimizes critical parameters including pH gradients in isoelectric focusing, flow rates in molecular sieve chromatography, and temperature control during processing. By carefully adjusting these parameters, the system achieves maximum purity (above 98.5%) with optimized processing times, resolving the contradiction between manufacturing precision and time loss

Inventive Principle:
Principle #35Parameter changes

3Measurement precision

If multiple characterization tests and uniformity tests are conducted, then the certification accuracy is ensured, but the time and resource consumption increase

Engineering Contradiction:
Improvecertification accuracyVSAvoidcertification efficiency
Core Design Contradiction:
Measurement precisionVSProductivity

Solution Approach 1:

The certification process employs a multi-functional testing protocol where a single set of experiments simultaneously determines multiple parameters. For example, the uniformity testing across different batches and storage conditions concurrently assesses purity, concentration accuracy, and stability, thereby ensuring comprehensive certification accuracy without proportionally increasing time and resource consumption

Inventive Principle:
Principle #6Universality (Multi-functionality)

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 provides a reliable reference material that ensures the accuracy and consistency of nCOV N protein measurements, enhancing the reliability of COVID-19 diagnosis and facilitating effective epidemic control.

Implementation Method 1

the biochemical reagent is purified by an isoelectric focusing method

Methodology Applied
Scientific EffectIsoelectric focusing: Isoelectric Focusing

Implementation Method 2

and/or a molecular sieve method to give the candidate nCOV N protein reference material with a purity of 98.5% or above

Methodology Applied
Scientific EffectMolecular sieve: Molecular Sieve

Implementation Method 3

an amino acid content in the hydrolysate is measured by performance liquid chromatography-isotopic dilution mass spectrometry (HPLC-IDMS)

Methodology Applied
Scientific EffectMass spectrometry:

Implementation Method 4

the mixture is hydrolyzed in an oven at 110° C. to 150° C. for 24 to 72 hours to give a hydrolysate

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentUS20240199707A1Method for preparing and certifying novel coronavirus nucleocapsid protein
Publication Date: 2024.06.20 NATIONAL INSTITUTE OF METROLOGY CHINA

AI summary

Disclosed in the present invention is a method for preparing and certifying a novel coronavirus nucleocapsid protein (nCOV N protein) reference material, the method including: purification and dilution of a raw material of the nCOV N protein; preliminary uniformity test and packaging of solution of a candidate nCOV N protein reference material; characterization of physicochemical properties of the solution of the candidate nCOV N protein reference material in the packaging units; uniformity and stability test of the nCOV N protein reference material in the packaging unit; measurement of standard N protein content of the nCOV N protein reference material, determination and statistical test of standard value of the nCOV N protein reference material; and uncertainty assessment of certified result of the standard value of the nCOV N protein reference material.