Anti-TNF Antibody Quantification via Labeled Peptide Mass Spectrometry
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Solution Overview
Problem
Current methods for quantifying anti-TNF antibodies in patients are limited by their specificity and sensitivity, particularly in distinguishing between different anti-TNF antibodies and handling samples with multiple antibodies, which complicates therapeutic monitoring and dosing adjustments.
Innovation Solution
A method involving the addition of labeled forms of anti-TNF antibodies to a sample, followed by enzyme proteolysis and mass spectrometric analysis to determine the ratio of labeled to native peptides, allowing for the quantification of anti-TNF antibodies regardless of the specific antibody type present, using peptides specific to Infliximab, Etanercept, Adalimumab, Certolizumab, and Golimumab.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If ELISA methods are used for quantifying anti-TNF antibodies, then sensitivity is improved, but reliability deteriorates due to inability to distinguish between different antibody types
Solution Approach 1:
The method segments the quantification process by first capturing all anti-TNF antibodies using a universal anti-Fc receptor, then separating and identifying specific antibody types (Infliximab, Etanercept, Adalimumab, Certolizumab, Golimumab) through targeted proteolysis and mass spectrometry analysis of unique peptide signatures for each antibody type
Solution Approach 2:
The invention introduces labeled forms of each specific anti-TNF antibody as internal standards that serve as intermediaries. These labeled antibodies are mixed with the patient sample, undergo identical proteolysis and detection processes, and provide reference signals for accurate quantification of each antibody type, thereby improving reliability
2Reliability
If mobility-shift assay based on high performance liquid chromatography is used, then specificity is improved, but sensitivity deteriorates due to low detection capability
Solution Approach 1:
The invention merges the advantages of both methods by combining the specificity approach of mobility-shift assay with the high sensitivity of mass spectrometry. The universal capture step ensures all antibodies are collected (sensitivity), while the subsequent type-specific proteolysis and MS analysis provides precise identification and quantification (specificity)
Solution Approach 2:
The method changes the detection parameter from optical detection (in chromatography) to mass spectrometric detection, which operates in a different parameter space with superior sensitivity. The use of labeled internal standards further optimizes the detection parameters for maximum sensitivity while maintaining specificity
3Adaptability or versatility
If multiple labeled forms of anti-TNF antibodies are added for comprehensive coverage, then versatility is improved, but device complexity increases
Solution Approach 1:
The invention achieves universality through a single universal anti-Fc capture receptor that can bind all anti-TNF antibody types, eliminating the need for multiple separate assays. This universal capture step simplifies the overall process while the subsequent mass spectrometry analysis handles the differentiation of specific antibody types, thereby improving versatility without proportionally increasing complexity
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 enables precise and accurate quantification of anti-TNF antibodies in human samples, regardless of the administered antibody, facilitating individualized dosing and monitoring across various treatment scenarios, including combination therapies.
Implementation Method 1
adding to a test sample which may contain therapeutic anti-TNF antibodies a known amount of two or more labeled forms of said anti-TNF antibodies
Implementation Method 2
subjecting the pre-proteolysis sample to an enzyme proteolysis, so as to provide a proteolysis sample comprising (i) proteolysis labeled peptides derived from the labeled anti-TNF antibodies and (ii) proteolysis peptides derived from the anti-TNF antibody contained in the test sample
Implementation Method 3
determining by mass spectrometric analysis the ratio between (i) one or more selected proteolysis labeled peptides and (ii) one or more corresponding proteolysis peptides derived from the said anti-TNF antibody
Data Source
AI summary
The present invention relates to a method for quantifying an anti-TNF antibody in a sample of a human individual comprising a step of adding to a test sample which may contain therapeutic anti-TNF antibodies a known amount of two or more labeled forms of said anti-TNF antibodies.