PerCP Stabilization in Antibody Formulations for Flow Cytometry
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Peridinin chlorophyll protein (PerCP) fluorochromes used in flow cytometry are inherently unstable, requiring refrigeration and are prone to rapid decay at higher temperatures, which can lead to experimental errors and delays in medical diagnosis or prognosis due to degradation during transport or storage.
Innovation Solution
A stable formulation comprising a labeled binding agent with a conjugated fluorochrome and unconjugated PerCP or its fragment, maintaining a total PerCP concentration of at least 0.1 μg/mL, which enhances stability without compromising fluorescence properties, allowing storage at temperatures above 8°C and reducing degradation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If PerCP fluorochrome is stored at temperatures above 8°C, then ease of operation and transport is improved, but the fluorochrome rapidly decays and loses stability
Solution Approach 1:
The patent introduces an intermediary substance (e.g., bovine serum albumin, sucrose, or other stabilizing agents) that mediates between the PerCP fluorochrome and the storage environment. This intermediary protects the PerCP from thermal degradation while allowing storage at higher temperatures, thus resolving the contradiction between ease of transport and fluorochrome stability.
Solution Approach 2:
The patent changes the chemical and physical parameters of the storage formulation by adjusting pH, ionic strength, and adding protective agents. These parameter changes create a stable environment for PerCP at elevated temperatures, enabling storage above 8°C without rapid decay while maintaining fluorochrome reliability.
2Loss of time
If PerCP formulation is transported without refrigeration, then loss of time and productivity is reduced, but the formulation decomposes and fluorescence intensity decreases
Solution Approach 1:
The patent applies preliminary action by pre-formulating PerCP with stabilizing agents and protective components before transport. This preliminary stabilization allows the formulation to withstand temperature fluctuations during transport without decomposition, reducing both transport time constraints and fluorochrome degradation.
Solution Approach 2:
The patent provides beforehand cushioning by incorporating protective substances that cushion the PerCP fluorochrome against thermal stress during transport. This prior protection prevents decomposition and maintains fluorescence intensity even when refrigeration is not available, thus reducing transport time losses without substance degradation.
3Measurement precision
If multiple fluorochromes are used in multicolor flow cytometry, then measurement precision and cell characterization capability are improved, but light sensitivity increases and requires minimization of light exposure
Solution Approach 1:
The patent applies local quality by optimizing the formulation conditions specifically for PerCP's light sensitivity while maintaining compatibility with other fluorochromes. Different stabilizing agents and buffer compositions are used locally tailored to PerCP's requirements, allowing multicolor flow cytometry with improved measurement precision while managing light sensitivity through localized formulation optimization.
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 formulation maintains fluorescence intensity stability by 25% or less over 1 day to 180 days at 20°C, enabling safe handling and storage at elevated temperatures, thus preventing premature decay and ensuring consistent results in flow cytometry analyses.
Implementation Method 1
PerCP has an extremely high extinction coefficient, a high quantum efficiency and a large Stokes shift. The peridinin molecules of PerCP absorb light in the blue-green wavelengths (470 to 550 nm) and transfer energy to the chlorophyll molecules with high efficiency.
Data Source
AI summary
The present disclosure is directed to formulations and methods that can be used for flow cytometry analysis. The present formulations and methods involve the use of PerCP to stabilize fluorochromes used in flow cytometry, allowing for use and storage of formulations at elevated temperatures and/or over long periods of time.