FOLR1 Detection via LC/MS Peptide Quantification
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Solution Overview
Problem
Current methods for detecting human folate receptor 1 (FOLR1) in samples are not sufficiently specific, suffer from competitive inhibition, and lack sensitivity, leading to inaccurate results and unreliable kits, particularly in cancer diagnostics where ovarian cancer often goes undetected until advanced stages.
Innovation Solution
A method involving immunocapture of FOLR1 using specific antibodies bound to a solid support, followed by digestion and liquid chromatography-mass spectrometry (LC/MS) analysis to quantify FOLR1 levels, using peptides that are not competitively inhibited by other antibodies or folic acid, allowing for precise detection even in complex samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional ELISA-based assays are used to detect FOLR1, then the assay can be performed with standard equipment, but the sensitivity is insufficient to distinguish between variations in shed FOLR1 at physiological levels and false positive results occur
Solution Approach 1:
The patent replaces traditional ELISA-based mechanical/chemical detection systems with mass spectrometry detection. The LC/MS system identifies FOLR1 through its unique mass-to-charge ratio of peptide fragments, providing superior sensitivity and specificity without relying on antibody-antigen binding that suffers from competitive inhibition and false positives.
Solution Approach 2:
The patent introduces liquid chromatography as an intermediary separation step between sample preparation and mass spectrometry detection. The chromatography system separates FOLR1 peptide fragments from other plasma components based on their hydrophobicity, enabling clean detection of target peptides at physiological concentrations without interference from the complex plasma matrix.
2Measurement precision
If assays use antibodies that bind to FOLR1, then FOLR1 can be detected, but competitive effects between antibody therapy and diagnostic antibody reduce accuracy
Solution Approach 1:
The patent eliminates antibody-based detection entirely and replaces it with mass spectrometry detection of peptide fragments. This substitution removes the source of competitive inhibition that occurs when therapeutic antibodies bind to FOLR1 and block diagnostic antibody access, allowing accurate detection regardless of therapy status.
Solution Approach 2:
The patent extracts FOLR1 from its native form and detects it through its constituent peptide fragments after proteolytic digestion. By detecting the mass-to-charge ratio of specific peptides rather than relying on intact protein-antibody binding, the assay becomes insensitive to the presence of therapeutic antibodies that would otherwise interfere with detection.
3Measurement precision
If assays require pre-treatment with light acid wash to dissociate folic acid from the receptor, then folic acid interference is reduced, but the procedure becomes more complex and time-consuming
Solution Approach 1:
The patent extracts FOLR1 from the plasma sample through immunocapture, then digests it into peptide fragments. By detecting specific peptide sequences rather than intact FOLR1 bound to folic acid, the method eliminates the need for acid wash steps to dissociate folic acid, as the peptide detection is not affected by folic acid binding.
Solution Approach 2:
The patent performs immunocapture of FOLR1 from plasma as a preliminary step before digestion and detection. This capture step selectively isolates FOLR1 from the complex plasma matrix, enriching the target analyte and eliminating the need for subsequent acid treatment to remove folic acid interference, thereby simplifying the overall workflow.
4Measurement precision
If many commercially available kits are used, then detection can be performed, but they are unreliable in reagents and lot-to-lot stability
Solution Approach 1:
The patent replaces antibody-based detection systems with mass spectrometry detection. This substitution eliminates variability in antibody reagents across different lots and manufacturers, as mass spectrometry detects the intrinsic mass-to-charge ratio of peptide fragments rather than relying on biological reagents that exhibit lot-to-lot variability.
Solution Approach 2:
The patent changes the detection parameter from antibody binding affinity (which varies between lots) to the mass-to-charge ratio of peptide fragments (which is an invariant physical property). This parameter change ensures consistent detection across different samples and laboratories without dependence on commercial kit reagents.
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 highly sensitive and accurate detection of FOLR1, enhancing diagnostic capabilities for cancers like ovarian cancer by overcoming previous limitations of specificity and sensitivity, enabling early detection and improved therapeutic strategies.
Implementation Method 1
capturing said folate receptor 1 (FOLR1) with an immunocapture reagent bound to a solid support
Implementation Method 2
performing liquid chromatography-mass spectrometry (LC/MS) analysis on the digested FOLR1
Implementation Method 3
performing liquid chromatography-mass spectrometry (LC/MS) analysis on the digested FOLR1, wherein said FOLR1 is detected by monitoring the chromatographic separation and mass spectrometric response of at least one signature FOLR1 peptide
Data Source
AI summary
The invention generally relates to methods and kits for the detection of human folate receptor 1 in a sample. Peptides of human folate receptor 1 are further provided.