Contributions
Abstract: PB1596
Type: Publication Only
Background
Traditional methods of diagnostic of hematologic cancers (flow cytometer, immunohistochemistry analysis, etc.) have a high sensitivity and specificity, but they require expensive equipment and specially trained staff, which increase the costs of these analysis. Therefore, the actual task is to develop a new tool – diagnostic biosensors which are expanding beyond traditional clinical labs to point-of-care and home settings. Nowadays zinc oxide (ZnO)-based nanostructures owing to unique physical properties - high photoluminescence (PL), biocompatibility and other multifunctional characteristics attract attention as building blocks for biosensor development. These properties of ZnO help retain biological activity of the immobilized biomolecule and help in achieving enhanced sensing performance.
Aims
In current work, we demonstrate the possibility of a fluorescent detection of human leukemic cells: T- and B-lymphoblasts, using zinc oxide nanorod (ZnO NR) platforms and immobilized on them specialized monoclonal antibodies (MABs) against cluster of differentiation (CD) proteins on the surface of investigated cancer cells (CD5 and СD19).
Methods
It was used human cell lines MOLT-4 derived from the patient with an acute lymphoblastic leukemia in relapse and IM-9 derived from the patient with a multiple myeloma and also healthy human’s peripheral mononuclear cells as control samples where expression of CD5 and CD19 antigens were found in 8-12% and 5-9% of the cell population, respectively.
For platform preparation, glass substrate was cleaned in ethanol and mQ and treated by the use of plasma technology; than ZnO NRs stock solution was dropped on it and annealed at 300°C.
Results
Firstly, we established the optimal concentrations of human anti-CD19 and anti-CD5 and their isotype controls (mouse anti-human IgG1 and IgG2a) for immune biosensor development, i.e. an appropriate amount of MABs to provide significant coverage of the ZnO NRs surface and maximum response of ZnO NRs photoluminescence. Next, it was shown that B- or T-lymphoblastoid cells bind to CD19 or CD5 targered ZnO NRs with high selectivity and PL signal increased on 50–70% in comparison with the signal from the control samples. Furthermore, rise of the ZnO NRs photoluminescence intensity correlated with the amount of CD19+ and CD5+ cells in the investigated populations (controlled using flow cytometry).
Conclusion
The outcomes of our study confirmed that ZnO NRs exhibit an optical property useful for effective monitoring of fluorescent signal from biological systems: human leukemic lymphocytes conjugated with CD19 or CD5 MABs, even at extremely low cell concentrations – from 5-10 till 250 cells per 1 mm2 of ZnO NRs platform. We propose that MABs-targeted ZnO NRs can be used for the development of biosensors for detection of human leukemic cells.
This work has received funding from the European Union’s Horizon 2020 research and innovation programme H2020-MSCA-RISE under grant agreement № 777926.
Session topic: 1. Acute lymphoblastic leukemia – Biology & Translational Research
Keyword(s): Antibody targeting, Diagnosis, Leukemia cell line, Molecular response
Abstract: PB1596
Type: Publication Only
Background
Traditional methods of diagnostic of hematologic cancers (flow cytometer, immunohistochemistry analysis, etc.) have a high sensitivity and specificity, but they require expensive equipment and specially trained staff, which increase the costs of these analysis. Therefore, the actual task is to develop a new tool – diagnostic biosensors which are expanding beyond traditional clinical labs to point-of-care and home settings. Nowadays zinc oxide (ZnO)-based nanostructures owing to unique physical properties - high photoluminescence (PL), biocompatibility and other multifunctional characteristics attract attention as building blocks for biosensor development. These properties of ZnO help retain biological activity of the immobilized biomolecule and help in achieving enhanced sensing performance.
Aims
In current work, we demonstrate the possibility of a fluorescent detection of human leukemic cells: T- and B-lymphoblasts, using zinc oxide nanorod (ZnO NR) platforms and immobilized on them specialized monoclonal antibodies (MABs) against cluster of differentiation (CD) proteins on the surface of investigated cancer cells (CD5 and СD19).
Methods
It was used human cell lines MOLT-4 derived from the patient with an acute lymphoblastic leukemia in relapse and IM-9 derived from the patient with a multiple myeloma and also healthy human’s peripheral mononuclear cells as control samples where expression of CD5 and CD19 antigens were found in 8-12% and 5-9% of the cell population, respectively.
For platform preparation, glass substrate was cleaned in ethanol and mQ and treated by the use of plasma technology; than ZnO NRs stock solution was dropped on it and annealed at 300°C.
Results
Firstly, we established the optimal concentrations of human anti-CD19 and anti-CD5 and their isotype controls (mouse anti-human IgG1 and IgG2a) for immune biosensor development, i.e. an appropriate amount of MABs to provide significant coverage of the ZnO NRs surface and maximum response of ZnO NRs photoluminescence. Next, it was shown that B- or T-lymphoblastoid cells bind to CD19 or CD5 targered ZnO NRs with high selectivity and PL signal increased on 50–70% in comparison with the signal from the control samples. Furthermore, rise of the ZnO NRs photoluminescence intensity correlated with the amount of CD19+ and CD5+ cells in the investigated populations (controlled using flow cytometry).
Conclusion
The outcomes of our study confirmed that ZnO NRs exhibit an optical property useful for effective monitoring of fluorescent signal from biological systems: human leukemic lymphocytes conjugated with CD19 or CD5 MABs, even at extremely low cell concentrations – from 5-10 till 250 cells per 1 mm2 of ZnO NRs platform. We propose that MABs-targeted ZnO NRs can be used for the development of biosensors for detection of human leukemic cells.
This work has received funding from the European Union’s Horizon 2020 research and innovation programme H2020-MSCA-RISE under grant agreement № 777926.
Session topic: 1. Acute lymphoblastic leukemia – Biology & Translational Research
Keyword(s): Antibody targeting, Diagnosis, Leukemia cell line, Molecular response