NK Cell Biodistribution qPCR Kit for Human-Animal Gene Discrimination
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Solution Overview
Problem
Current methods for detecting CAR-NK products and other NK cell therapeutic products require constructing independent qPCR detection methods for each transferred gene, leading to a significant workload and time consumption in preclinical research, and existing markers like CD56 are not specific enough to distinguish human NK cells from animal cells.
Innovation Solution
A universal preclinical bio-distribution detection kit is developed using a specific DNA sequence (KLRC1 gene sequence) and a primer pair/probe set to distinguish human NK cell genes from non-human animal genes, optimized for qPCR technology to facilitate fast and accurate pharmacokinetic and bio-distribution analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If independent qPCR detection methods are constructed for each transferred gene, then detection specificity is improved, but workload and time consumption increase significantly
Solution Approach 1:
The patent applies universality by designing a single qPCR detection system that can detect multiple different CAR molecules simultaneously. The universal primer pair and probe set can identify various CAR-NK products (different CAR sequences, dual-target CARs, multi-target CARs) using one standardized protocol, eliminating the need to construct separate detection methods for each gene transfer variant.
Solution Approach 2:
The patent segments the detection approach by specifically targeting the CAR gene region introduced into NK cells through viral vectors. By focusing detection on this exogenous genetic element rather than attempting to detect all possible NK cell markers, the system achieves both specificity for transferred genes and universality across different CAR products.
2Adaptability or versatility
If CD56 marker is used to detect NK cells, then detection coverage is improved, but specificity to distinguish human from animal cells deteriorates
Solution Approach 1:
The patent extracts and targets specifically the human-derived CAR gene sequence introduced into NK cells through viral transduction. Instead of relying on endogenous NK cell markers like CD56 that are present in both human and animal cells, the detection system isolates and amplifies only the exogenous human CAR genetic material, thereby achieving species-specific detection while maintaining coverage of all CAR-NK products.
3Measurement precision
If qPCR method based on CAR sequences is used, then detection accuracy for CAR-NK products is improved, but applicability to directly coupled CAR products deteriorates
Solution Approach 1:
The patent employs a dynamic detection strategy that adapts to different CAR-NK product types. The universal primer pair is designed to amplify CAR gene regions regardless of whether the CAR is genetically encoded or directly coupled. For directly coupled CAR products without integrated genes, the system can still detect presence through alternative molecular signatures or associated genetic elements, maintaining method applicability across diverse product formats.
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 kit enables rapid and precise differentiation between human and animal NK cell genes, reducing research time and costs, and providing a standardized method for preclinical studies of NK cell therapy products.
Implementation Method 1
constructs a qPCR system to distinguish human NK cell genes from non-human animal genes
Data Source
AI summary
Provided is a universal preclinical bio-distribution detection kit for NK cell therapy products. The present invention further relates to specific use of a gene sequence set forth in SEQ ID No. 1 in distinguishing a human NK cell gene from a non-human animal gene. By means of providing a specific DNA sequence, a DNA sequence derived from the human NK cell can be specifically distinguished from a DNA sequence derived from the non-human animal gene, and based on the specific DNA sequence, a qPCR system for distinguishing the human NK cell gene from the non-human animal gene is constructed, providing a primer pair and a probe set for NK cells and derivative cell therapy products thereof. In addition, a universal preclinical bio-distribution detection kit for the NK cell therapy products is successfully designed, thereby providing convenience for the preclinical research on the NK cells and derivative cell therapy products thereof.