Conformation-Specific p53 Fragment Mass Spectrometry for Alzheimer’s
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for diagnosing and prognosing neurodegenerative diseases such as Alzheimer's disease and dementia lack sensitivity and specificity, particularly in detecting conformationally altered p53 protein isoforms that are indicative of these conditions.
Innovation Solution
The use of specific anti-p53 antibodies with defined CDR sequences to form immunocomplexes with conformationally altered p53 isoforms, followed by enzymatic digestion and mass spectrometry to quantify p53 peptide fragments, specifically P1 peptide (TEEENLR) for diagnosis and prognosis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional diagnostic methods are used for neurodegenerative diseases, then the diagnostic process is simple, but the sensitivity and specificity for detecting conformationally altered p53 protein isoforms are insufficient
Solution Approach 1:
The diagnostic method is segmented into distinct sequential steps: immunocomplex formation with conformation-specific antibodies, enzymatic digestion of captured p53 protein, and mass spectrometry analysis of resulting peptides. This segmentation allows each step to be optimized independently, achieving high sensitivity and specificity through the combination of conformational antibody binding followed by controlled proteolytic cleavage and precise mass measurement
Solution Approach 2:
Conformation-specific anti-p53 antibodies serve as intermediaries that selectively bind to and capture unfolded p53 isoforms from complex biological samples. These antibodies act as mediators between the target analyte (conformationally altered p53) and the detection system (mass spectrometry), enabling specific isolation and enrichment of the disease-relevant protein isoforms before analysis
2Measurement precision
If mass spectrometry is used to quantify p53 peptide fragments, then detection sensitivity is improved, but the measurement and detection difficulty increases
Solution Approach 1:
P53 protein is captured in its intact form using conformation-specific antibodies before digestion. This preliminary capture and enrichment step concentrates the target analyte and removes interfering substances from complex biological matrices, thereby simplifying subsequent peptide detection and improving mass spectrometry measurement accuracy
Solution Approach 2:
The method focuses measurement on specific localized regions of the p53 protein by using conformation-specific antibodies that bind to unique epitopes on unfolded p53. Following digestion, only peptides derived from the antibody-bound region are analyzed by mass spectrometry, concentrating the measurement on disease-relevant local structures rather than analyzing the entire protein
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
Provides accurate and sensitive detection of neurodegenerative diseases by quantifying p53 peptide fragments, allowing for early diagnosis and prognosis of Alzheimer's disease and dementia with high specificity and sensitivity.
Implementation Method 1
contacting the biological sample or a portion thereof of the subject with an anti-p53 antibody comprising heavy chain CDR sequences (CDR1 (SEQ ID NO:8), CDR2 (SEQ ID NO: 9) and CDR3 (SEQ ID NO: 10)) and light chain CDR sequences (CDR1 (SEQ ID NO:11), CDR2 (SEQ ID NO: 12) and CDR3 (SEQ ID NO: 13)) in a reaction mixture to form an immunocomplex comprising the p53 isoform and the antibody
Implementation Method 2
subjecting the eluted p53 isoform or a portion thereof to enzymatic digestion, thereby to generate a composition comprising one or more proteolytic peptides comprising P1 peptide (TEEENLR, SEQ ID NO: 1)
Implementation Method 3
quantifying an amount of the P1 peptide in the composition or a portion thereof by mass spectrometry, wherein the quantifying comprises determining the intensity of mass/charge ratio corresponding to at least one fragment of the P1 peptide
Data Source
AI summary
Disclosed are fragments of p53 peptide (P1) and their use in the diagnosis and/or prognosis of Alzheimer's disease (AD) in a biological sample. The invention provides a method based on mass spectrometry analysis for the diagnosis of Alzheimer's disease at the pre-clinical and prodromal stages of the disease and for the prognosis of cognitive decline in a subject, by quantitating the levels of one or more p53 peptide fragments in a biological sample of a subject.