Flow Cytometry Quantification of Virus-Size Particles Using Fluorescent Antibody Stains

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current methods for quantifying viral particles, especially in gene therapy and vaccine production, are unreliable due to high variability, time-consuming, and costly, leading to issues like product loss, delays, and adverse immune responses.

Innovation Solution

The method involves using flow cytometry with a fluorescent antibody stain that binds to specific epitopes on virus-size particles, allowing for the detection and quantification of unassociated virus-size particles without the need for particle identification by light scatter detection or separation from residual unbound stain.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If traditional methods (quantitative PCR, absorbance readings) are used to quantify viral particles, then the process can be performed, but the results are highly variable and unreliable

Engineering Contradiction:

Improvereliability of viral particle quantificationVSAvoidprecision of viral particle quantification

Core Design Contradiction:

ReliabilityVSMeasurement precision

Solution Approach 1:

The patent replaces traditional mechanical/chemical quantification methods (PCR, absorbance readings) with flow cytometry, which uses optical detection principles to directly count and characterize individual viral particles based on their light scattering properties and fluorescent staining, thereby improving both reliability and precision

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent employs fluorescent stains that bind to viral particles, causing them to emit light at specific wavelengths when excited. This color/fluorescence change enables direct optical detection and counting of viral particles, providing more reliable and precise quantification compared to traditional methods

Inventive Principle:

Principle #32Color changes

2Productivity

If conventional quantification methods are used, then viral particles can be detected, but the process is time-consuming and costly

Engineering Contradiction:

Improvespeed of viral particle quantificationVSAvoidtime required for quantification process

Core Design Contradiction:

ProductivityVSLoss of time

Solution Approach 1:

The patent replaces time-consuming traditional methods with flow cytometry, which can rapidly analyze thousands of particles per second through automated optical detection, significantly reducing quantification time while maintaining high accuracy

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The flow cytometry system automatically performs particle detection, counting, and characterization without requiring manual intervention, thereby increasing productivity and reducing the time and cost associated with conventional manual or semi-automatic methods

Inventive Principle:

Principle #25Self-service

3Ease of operation

If light scatter detection is used to identify particles, then particle detection is possible, but the method is complex and requires additional separation steps

Engineering Contradiction:

Improvesimplicity of particle detection processVSAvoidcomplexity of detection system

Core Design Contradiction:

Ease of operationVSDevice complexity

Solution Approach 1:

The patent uses flow cytometry to simultaneously perform multiple functions: particle detection, counting, size characterization, and fluorescent staining, all within a single integrated system, thereby simplifying the overall process and reducing the need for separate detection and separation steps

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

This approach provides a rapid, accurate, and cost-effective means of quantifying virus-size particles, even in complex samples with high background signals, thereby improving the reliability of viral particle detection and reducing the risks associated with viral vector production.

Implementation Method 1

a fluorescent antibody stain capable of binding, directly or indirectly, with the unassociated virus-size particles through an epitope

Methodology Applied

Scientific EffectAntigen-antibody binding:

Implementation Method 2

subjecting the fluid sample flowing through the flow cell to excitation radiation capable of causing a fluorescent emission response from the fluorescent antibody stain

Methodology Applied

Scientific EffectFluorescence: Fluorescence

Data Source

PatentUS12339283B2Flow cytometry evaluation of biological material for unassociated virus-size particles

Publication Date: 2025.06.24 SARTORIUS BIOANALYTICAL INSTRUMENTS INC

AI summary

A method for evaluating a biological material for unassociated virus-size particles having a particular epitope uses a fluorescent antibody stain specific for binding with the epitope and a fluid sample with the virus-size particles and fluorescent antibody stain is subjected to flow cytometry with identification of fluorescent emission detection events indicative of passage through a flow cell of a flow cytometer of unassociated labeled particles of virus size including such a virus-size particle and fluorescent antibody stain.