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What is Liquid Biopsy?  


    Cancer detection and monitoring method


    Liquid Biopsy, also known as fluid biopsy, is a significant branch of in vitro diagnosis that is primarily based on blood or other body fluid for disease analysis and monitoring, such as cancer. With a simple blood draw, circulating tumor biomarkers in the blood sample provide information on tumor progression and treatment efficacy, and guiding personalized precision medication direction. Compared with traditional tissue biopsy, liquid biopsy has provided additional benefits of real-time monitoring, avoid unrepresentative sampling in heterogenous tumor, and provides comprehensive clinical information.


    Liquid Biopsy is a branch of in vitro diagnostics that analyses and diagnoses diseases such as cancer through the use of body liquids like blood or urine. It was rated as one of the “Breakthrough Technologies 2015” by MIT Technology Review.


    Compared to traditional tissue biopsy, liquid biopsy is equipped with unique advantages like real-time monitoring, ability to overcome tumor heterogeneity, and comprehensive diagnostic information. Currently, in clinical researches, liquid biosy mainly includes tests for CTC (Circulating Tumor Cell), ctDNA (Circulating Tumor DNA), exosomes, and Circulating miRNA etc. In comparison to traditional methods like observation of clinical symptoms or diagnostic imaging, using liquid biopsy can predict the risk of cancer earlier, enabling “Early discovery, early screening, early treatment”.


What is Circulating miRNA?


microRNAs or miRNAs are small RNA molecules that are non-coding and have important effects on gene expression, often inhibitory.  By binding to mRNAs in the cells, the microRNAs inhibit DNA functioning, thus affecting important life functions in a cell. In many cancer types, miRNAs production is upregulated, converting an ordinary cell into a cancerous one. Additionally, these miRNAs can be transported outside the cell, in the tumour environment, in sacs/vesicles called 'exosomes'. The presence of a high number of exosomal miRNAs has been associated with the spread of certain types of cancer in the body. Isolation and detection of miRNAs from a patient’s blood can help in early cancer diagnosis, without the need for invasive surgeries.

What is Circulating Tumor Cell (CTC)?


Circulating tumor cell (CTCs) are the cancer cells that have detached from the primary tumor or metastatic tumor into circulatory system during tumor progression or metastases. CTC provides comprehensive information of cancer in cellular, genetics and protein levels. Hence, CTC detection is an effective tool for earlier detection, evaluation of therapeutic efficacy, individual precision medicine selection, drug resistance detection, earlier tumor recurrence evaluation and real-time monitoring of cancer.


How can we detect CTCs in blood?


Cellomics CTC detection Platform - New generation of CTC viable Cell Sorting.


The CTC detection platform has employed the leading microfluidic chip technology that select CTCs extensively by cell size, cell shape and multiple surface biomarkers. This technology has combined the physical, chemical and biological properties of CTCs to uphold the high sensitivity, high specificity and high accuracy of CTC detection.


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 CTC counting : 


  • According to current studies, CTC is recognized as prognostic factor for lots of cancers, including breast cancer, prostate cancer, bowel cancer and more. The higher number of CTC in blood indicates poor prognosis of patients. Comparing with conventional radiological examination, CTC counting affords much more prior cancer screening and auxiliary diagnosis, and provides more authentic total survival time of patients.




Example:


  • The number of CTC >5 demonstrated lower survival rate than the number of CTC <5 in 7.5mL peripheral blood from breast cancer patients and bowel cancer patients respectively.

 CTC tumor classification : 

 

  • “Epithelial-Mesenchymal transition” means the process of CTC inter-conversion between cancer-cell states involving epithelial, mesenchymal and epithelial-mesenchymal when it enters the blood circulating system. It is important to note that mesenchymal CTCs have higher possibilities of migration. 
  • To classify CTC tumor and accelerate tumor determination, CELLOMICS CTC100 cell sorter makes the isolated CTCs perfectly suitable for further downstream analysis. The sorted cancer cells can be visualized by immunostaining; the fluorescent-tagged immuno-protein can specifically bind to epithelial biomarker EpCam or mesenchymal biomarker Cell Surface Vimentin (CSV) on cancer cells and reflect light to anticipate the cell membrane of CTC.

 


 

Example:

 

  • CTC with EpCam expression show more clear cancer progression

 Immunostaining : 

 

  • Other than tumor classification, CELLOMICS CTC100 cell sorter and immunostaining can also be applied on CTC target protein expression detection (such as PD-L1 expression and etc.), which provides useful information for medical treatment selection and dynamic monitoring and etc.

 


 

Example:

 

  • PD-1/PD-1 immunotherapy is the most prevalent anti-cancer therapy nowadays. It aims at triggering natural immune responses to defense cancer. Despite years of clinical verification, it does not applicable to all cancer patients. Consequently, it raises the necessity to deal with the problem of sensitive people determination. According to studies, highly expressed PD-L1 in CTC patients may have higher chance to be benefit from PD-1 immunotherapy.
  • -  PD-L1 sensitive test before treatment to evaluate the fitness of PD-L1 to individuals
  • -  PD-L1 immunotherapy therapeutic effect real time monitoring

 Immuno-FISH technique : 

 

  • CTC that is enriched by CELLOMICS CTC100 Cell Sorter is able to perform Fluorescence in situ hybridization (FISH) technique for DNA mutation detection. The fluorescent-tag-probes hybridize to particular gene location and reveal the position and number of CTC mutation sites, including EML4-ALK, HER2, cMET and etc. CTC immuno-FISH technique contributes to improve current CTC identification by molecular properties and provides more reliable information for CTC classification.



 Digital PCR and gene targeting test : 


  • The CTC samples collect from CELLOMICS CTC100 Cell Sorter can be employed in PCR-based gene targeting analysis for instance, droplet digital PCR (ddPCR), quantitative PCR (qPCR) and etc. The detecting genes contain lung cancer or breast cancer mutation genes EGFR, KRAS, TP53, PIK3CA and etc. They are crucially relevant to tumor targeting therapy for treatment guide and real-time monitoring by providing comprehensive and personalized genomic information for patient.




Example:


  • -  Detecting EGFR mutation in CTC with ddPCR

 NGS and Driver gene detection : 

 

  • CELLOMICS CTC100 Cell Sorter is also able to sort out CTC for Next Generation Sequencing (NGS) and Single Cell Sequencing. Integrating the whole genome sequencing, exosomal sequencing, targeted gene sequencing, transcriptome sequencing and methylation sequencing, can the CTC molecular mechanism and metastatic mechanism be studied and analyzed thoroughly, and uncovered unknown gene targets.

  • Through sequencing, it suggested that MET-CTC and EMT-CTC demonstrated enormous differences in MET, EMTHE and CSC-related gene expression. 

 

 Cell Culture/PDX : 

 

  • An intact CTC carries a lot of tumor information, including proteins, DNA mutation, coding DNA, non-coding DNA and etc. In vitro CTC culturing for drug sensitivity test has been proved to be applicable to medication selections, which exhibits foreseeable clinical potentials.
  • (Consensus among professionals: Liquid Biopsy in clinical tumor treatment applications and practices of medical diagnosis)
  • -  In vitro Breast Cancer CTC multiplication
  • -  The drug sensitivity test has manifested the same results with clinical cases


 CTC counting : 


  • According to current studies, CTC is recognized as prognostic factor for lots of cancers, including breast cancer, prostate cancer, bowel cancer and more. The higher number of CTC in blood indicates poor prognosis of patients. Comparing with conventional radiological examination, CTC counting affords much more prior cancer screening and auxiliary diagnosis, and provides more authentic total survival time of patients.




Example:


  • The number of CTC >5 demonstrated lower survival rate than the number of CTC <5 in 7.5mL peripheral blood from breast cancer patients and bowel cancer patients respectively.

 CTC tumor classification : 

 

  • “Epithelial-Mesenchymal transition” means the process of CTC inter-conversion between cancer-cell states involving epithelial, mesenchymal and epithelial-mesenchymal when it enters the blood circulating system. It is important to note that mesenchymal CTCs have higher possibilities of migration. 
  • To classify CTC tumor and accelerate tumor determination, CELLOMICS CTC100 cell sorter makes the isolated CTCs perfectly suitable for further downstream analysis. The sorted cancer cells can be visualized by immunostaining; the fluorescent-tagged immuno-protein can specifically bind to epithelial biomarker EpCam or mesenchymal biomarker Cell Surface Vimentin (CSV) on cancer cells and reflect light to anticipate the cell membrane of CTC.

 


 

Example:

 

  • CTC with EpCam expression show more clear cancer progression

 CTC counting : 


  • According to current studies, CTC is recognized as prognostic factor for lots of cancers, including breast cancer, prostate cancer, bowel cancer and more. The higher number of CTC in blood indicates poor prognosis of patients. Comparing with conventional radiological examination, CTC counting affords much more prior cancer screening and auxiliary diagnosis, and provides more authentic total survival time of patients.




Example:


  • The number of CTC >5 demonstrated lower survival rate than the number of CTC <5 in 7.5mL peripheral blood from breast cancer patients and bowel cancer patients respectively.

 CTC tumor classification : 

 

  • “Epithelial-Mesenchymal transition” means the process of CTC inter-conversion between cancer-cell states involving epithelial, mesenchymal and epithelial-mesenchymal when it enters the blood circulating system. It is important to note that mesenchymal CTCs have higher possibilities of migration. 
  • To classify CTC tumor and accelerate tumor determination, CELLOMICS CTC100 cell sorter makes the isolated CTCs perfectly suitable for further downstream analysis. The sorted cancer cells can be visualized by immunostaining; the fluorescent-tagged immuno-protein can specifically bind to epithelial biomarker EpCam or mesenchymal biomarker Cell Surface Vimentin (CSV) on cancer cells and reflect light to anticipate the cell membrane of CTC.

 


 

Example:

 

  • CTC with EpCam expression show more clear cancer progression

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