TEPATM TECHNOLOGY PLATFORM
TEPATM is proprietary TARGETED ENRICHMENT AND PREAMPLIFICATION technology.
It solves signal-to-noise problem in liquid biopsy by increasing detection levels of mutant cfDNA in blood up to 1000 times. This crucial improvement allows liquid biopsy to finally become a viable tool in early cancer detection.
CHALLENGES AND PROBLEMS IN LIQUID BIOPSY
LOW CONCENTRATION Amount of cfDNA from tumors in early stages of cancer is less than 0.1% of the total circulating DNA in plasma
FRAGMENTATION cfDNA from tumors is highly fragmented and shorter than cfDNA from normal cells (Underhill et al, 2016)
LIMITATIONS OF cfDNA ISOLATION TECHNIQUES cfDNA isolation kits currently available on the market are biased towards longer DNA fragments and higher GC content. DNA fragments shorter than 75bp are lost during isolation and cannot be accessible for NGS analysis.
LOW SENSITIVITY Sensitivity of currently available qPCR/NGS based methods is insufficient for detecting low content of mutant cfDNA
1. Isolation of cfDNA including extra-short fragments (20-50 bp) using proprietary SubXTM technology with no bias for DNA length and GC content.
2. Enrichment of mutant cfDNA by: a- elongation of short fragments and b- selective preamplification of mutant copies up to 1000 times to the level of reliable detection.
Selective preamplification of mutant cfDNA fragments leading to a 1000 fold increase in qPCR and NGS sensitivity.
ADDITIONAL CLINICAL APPLICATION
Reliable estimation of the tumor specific DNA content vs normal (WT) DNA for tumor mutation burden analysis
Multiplexing for detection of mutations with clinical relevance
Multianalyte analysis (e.g. exoDNA vs cfDNA)
Applicable to other bioliquids - urine, CSF, saliva, etc.