Multi-Protein Quantitation Standard for Single-Lane Gel Electrophoresis
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Solution Overview
Problem
Current methods for quantifying proteins in gel electrophoresis require generating multiple standard curves using separate lanes for each dilution series, limiting available lanes for analyzing target proteins and being restricted to pure protein solutions.
Innovation Solution
A protein quantitation standard comprising a mixture of unstained and prestained polypeptides with varying molecular weights and electrophoretic mobilities, allowing for simultaneous generation of a molecular weight ladder and mass standard curve in a single lane, enabling protein quantification without the need for multiple lanes or purified protein solutions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If separate lanes are used for each dilution series of protein standards, then accurate protein quantification can be achieved, but the number of available lanes for analyzing target proteins is reduced
Solution Approach 1:
The patent combines multiple protein standards with different known masses into a single mixture that can be loaded into one lane. This mixture contains proteins with varying molecular weights and concentrations, allowing the generation of a standard curve from a single lane instead of requiring separate lanes for each dilution series.
Solution Approach 2:
The protein standard mixture serves multiple functions simultaneously: it provides reference points for quantification, spans a range of molecular weights, and enables standard curve generation all within a single lane, replacing the need for multiple specialized lanes.
2Measurement precision
If purified protein solutions are used for quantification, then accurate mass determination can be achieved, but the method is restricted to pure protein samples only
Solution Approach 1:
The method is designed to work with both purified protein samples and complex mixtures containing multiple proteins. The protein standard mixture provides reference points that can be compared against various sample types, making the quantification method universally applicable rather than limited to pure proteins only.
Solution Approach 2:
The patent changes the parameter of sample purity requirements by using a mixture of protein standards with different molecular weights and concentrations. This allows the method to accommodate variations in sample complexity while maintaining quantification accuracy through comparison with the multi-component standard.
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
Enables efficient and accurate quantification of protein mass and molecular weight in a single lane of a gel, increasing lane availability for analyzing target proteins and allowing for protein quantification in both denaturing and non-denaturing gels.
Implementation Method 1
Gel electrophoresis, including polyacrylamide gel electrophoresis (PAGE), is a common laboratory technique for estimating the molecular weight of proteins and polypeptides. Gel electrophoresis is used to separate proteins based on their charge, size and/or molecular weight.
Implementation Method 2
Under denaturing conditions, for example, in the presence of an anionic detergent such as sodium dodecyl sulphate (SDS), proteins can be separated based on their molecular weights. The anionic detergent both unfolds the protein and provides a uniform negative charge density such that the electrophoretic mobility of the protein is a linear function of the logarithm of the molecular weight.
Data Source
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AI summary
Provided is a protein quantitation standard having polypeptides of different molecular weights that functions as a mass quantitation standard in one lane of an electrophoretic gel. The protein quantitation standard includes unstained polypeptides having different electrophoretic mobilities that are present in different quantities, such that the bands in the gel have different intensities when visualized. The protein quantitation standard can also contain prestained polypeptides that function as visual molecular weight markers. Also provided are methods of determining the mass of a target polypeptide or protein using the protein quantitation standard.