Multiplex Primer Panel for Tumor Immune Response Assessment
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Solution Overview
Problem
Current methods for identifying responsive candidates to cancer immunotherapies and monitoring response are inefficient and highly variable, particularly due to limitations in measuring PD-L1 protein levels using immunohistochemistry, necessitating a higher-throughput, systematic, and standardized solution for understanding tumor microenvironment and drug response biomarkers.
Innovation Solution
A multiplex assay using primer pair reagents targeting immune response genes and housekeeping genes to measure expression levels in biological samples, covering checkpoint pathways, T cell signaling, tumor markers, and other relevant functions, allowing for the determination of immune response activity through multiplex amplification and detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If immunohistochemistry is used to measure PD-L1 protein levels, then PD-L1 measurement is achieved, but the method is inefficient and highly variable
Solution Approach 1:
The patent replaces the mechanical immunohistochemistry method with a molecular biology-based qPCR system. Instead of using antibodies and protein detection mechanisms, the invention uses primer pairs to amplify and quantify PD-L1 mRNA transcripts, substituting a biochemical amplification system for the immunological detection system, thereby achieving higher precision and throughput
Solution Approach 2:
The patent changes the detection parameter from protein level measurement (immunohistochemistry) to mRNA expression level measurement (qPCR). This parameter transformation allows for more sensitive and quantitative detection of PD-L1 expression, resolving the contradiction between measurement precision and throughput efficiency
2Reliability
If immunohistochemistry is used to measure PD-L1 protein levels, then PD-L1 measurement is achieved, but the method is highly variable
Solution Approach 1:
The patent replaces the variable immunohistochemistry method with a standardized qPCR system. The molecular amplification process provides more consistent and reproducible results compared to antibody-based staining, improving reliability while the standardized protocol reduces operational complexity
Solution Approach 2:
The patent uses mRNA copying and amplification through PCR to generate multiple copies of the target sequence. This amplification process creates sufficient signal for detection while providing standardized, reproducible measurements that improve reliability across different samples and laboratories
3Adaptability or versatility
If a single marker (PD-L1) is measured, then simple detection is achieved, but comprehensive understanding of tumor microenvironment is limited
Solution Approach 1:
The patent creates a multiplex qPCR assay that simultaneously measures multiple immune checkpoint genes (PD-L1, PD-1, CTLA-4, LAG-3, TIM-3) and housekeeping genes in a single reaction system. This multi-functional assay provides comprehensive tumor microenvironment assessment while maintaining operational efficiency through standardized protocols
Solution Approach 2:
The patent combines multiple gene detection functions into a single qPCR assay system. By merging the detection of multiple immune checkpoint markers and normalization controls into one unified platform, the invention achieves comprehensive assessment capability without proportionally increasing operational 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
The solution provides a robust and specific method for biomarker screening and understanding tumor immune response mechanisms, offering a comprehensive picture of the tumor microenvironment and improving the identification of responsive candidates and monitoring of treatment responses.
Implementation Method 1
Amplification comprises contacting at least a portion of the sample comprising multiple target sequences of interest with the composition of the invention in the presence of a polymerase under amplification conditions to produce a plurality of amplified target expression sequences
Data Source
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AI summary
The present invention provides methods and compositions that are useful for assessing gene expression for tumor immune response profile of a sample. In particular, a target-specific primer panel is provided that allows for selective amplification of immune response target sequences in a sample. In one aspect, the invention relates to target-specific primers useful for selective amplification of one or more target sequences associated with immune response. In some aspects, amplified target sequences obtained using the disclosed methods, and compositions can be used in various processes including nucleic acid sequencing and used to detect the presence of genetic variants and/or expression levels of one or more targeted sequences associated with immune response.