A strain-promoted alkyne–azide cycloaddition (SPAAC) reaction of a novel EpCAM aptamer–fluorescent conjugate for imaging of cancer cells

Subramanian, Nithya; Sreemanthula, Jagadeesh Babu; Balaji, B.; Kanwar, Jagat R.; Biswas, Jyotirmay; Krishnakumar, Subramanian

doi:10.1039/c4cc02996h

Cited by 21 publications

(14 citation statements)

References 26 publications

(25 reference statements)

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“…In particular, aptamer SYL3C was identified and is currently widely used in the elucidation of cancer clinical samples, 53 immunostaining of cancer frozen tissues, 54 and imaging of cancer cells. 55 …”

Section: Aptamer-based Molecular Recognition For Biomarker Targetingmentioning

confidence: 99%

Molecular Elucidation of Disease Biomarkers at the Interface of Chemistry and Biology

et al. 2017

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Disease-related biomarkers are objectively measurable molecular signatures of physiological status that can serve as disease indicators or drug targets in clinical diagnosis and therapy, thus acting as a tool in support of personalized medicine. For example, the prostate-specific antigen (PSA) biomarker is now widely used to screen patients for prostate cancer. However, few such biomarkers are currently available, and the process of biomarker identification and validation is prolonged and complicated by inefficient methods of discovery and few reliable analytical platforms. Therefore, in this Perspective, we look at the advanced chemistry of aptamer molecules and their significant role as molecular probes in biomarker studies. As a special class of functional nucleic acids evolved from an iterative technology termed Systematic Evolution of Ligands by Exponential Enrichment (SELEX), these single-stranded oligonucleotides can recognize their respective targets with selectivity and affinity comparable to those of protein antibodies. Because of their fast turnaround time and exceptional chemical properties, aptamer probes can serve as novel molecular tools for biomarker investigations, particularly in assisting identification of new disease-related biomarkers. More importantly, aptamers are able to recognize biomarkers from complex biological environments such as blood serum and cell surfaces, which can provide direct evidence for further clinical applications. This Perspective highlights several major advancements of aptamer-based biomarker discovery strategies and their potential contribution to the practice of precision medicine.

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Section: Aptamer-based Molecular Recognition For Biomarker Targetingmentioning

confidence: 99%

Molecular Elucidation of Disease Biomarkers at the Interface of Chemistry and Biology

et al. 2017

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“…Scatter plot of total EpCAM/anti‐EpCAM bond number versus cell surface area further revealed that for MCF‐7 and PC3 cells, the total number of EpCAM/anti‐EpCAM bonds was linearly proportional to cell surface area; however, for MCF‐10A and HeLa cells, such linear correlation was not observable (Figure d). These observations further supported positive and comparable EpCAM expression for MCF‐7 and PC3 cells and negative EpCAM expression for MCF‐10A and HeLa cells . Assuming homogenous distribution of EpCAM on cancer cell membranes, average EpCAM/anti‐EpCAM bond density C bond for MCF‐10A, MCF‐7, PC3, and HeLa cells were 1.36 ± 0.62 bonds μm −2 , 29.71 ± 2.66 bonds μm −2 , 34.36 ± 3.94 bonds μm −2 , and 9.29 ± 1.57 bonds μm −2 , respectively.…”

Section: Resultsmentioning

confidence: 99%

“…We should note that the EpCAM/anti‐EpCAM bond quantities measured by the elasticity microcytometer were different from total EpCAM expression conventionally measured at the single cell level by immunostaining or fluorescence‐activated cell sorting. Nonetheless, the total numbers of EpCAM/anti‐EpCAM bonds on MCF‐10A, MCF‐7, PC3, and HeLa cells determined from the elasticity microcytometer were consistent and on the same order of magnitude with the total EpCAM expression data reported in the literature …”

Section: Resultsmentioning

confidence: 99%

“…Nonetheless, the total numbers of EpCAM/anti-EpCAM bonds on MCF-10A, MCF-7, PC3, and HeLa cells determined from the elasticity microcytometer were consistent and on the same order of magnitude with the total EpCAM expression data reported in the literature. [ 63,64 ] 2D scatter plot of EpCAM/anti-EpCAM bond density C bond versus cell stiffness was generated to further compare cellular phenotypes between MCF-10A and MCF-7 cells ( Figure 7 ). Importantly, clustering of MCF-10A and MCF-7 cells in the scatter plot clearly revealed a signifi cant distinction between these two cell types with respect to EpCAM expression and cell stiffness.…”

Section: Quantitative Measurement Of Epcam Expression On Single Cancementioning

confidence: 99%

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Multiparametric Biomechanical and Biochemical Phenotypic Profiling of Single Cancer Cells Using an Elasticity Microcytometer

Chen

et al. 2016

Small

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Deep phenotyping of cancer cells at the single-cell level is of critical importance in the era of precision medicine to advance understanding of the precise relationship between gene mutation and cell phenotype and to elucidate biological nature of tumor heterogeneity and their potential biological and clinical implications. Existing microfluidic single-cell phenotyping tools, albeit their high-throughput, high-resolution operation, are limited to phenotypic measurements of 1 – 2 selected morphological and physiological features of single cells. To address the critical need for multiplexed, informative phenotyping of live single cancer cells, herein we reported a microfluidic elasticity microcytometer for multiparametric biomechanical and biochemical phenotypic profiling of free-floating, live single cancer cells to obtain quantitative information of cell size, cell deformability / stiffness, and expression levels of surface receptors simultaneously for the same single live cancer cells. The elasticity microcytometer was implemented for single-cell measurements and comparisons of four human cell lines with distinct metastatic potentials and derived from different human tissues. An analytical model was developed from first principles for the first time to effectively convert cell deformation and adhesion information of single cancer cells encapsulated inside the elasticity microcytometer to cell deformability / stiffness and surface protein expression. Together, the elasticity microcytometer holds a great promise for comprehensive molecular, cellular, and biomechanical phenotypic profiling of live cancer cells at the single cell level, critical for studying intra-tumor cellular and molecular heterogeneity using low-abundance, clinically relevant human cancer cells.

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“…Aptamers and inhibitors require additional steps before conjugation with imaging agents, such modification could change their binding affinity to receptors. 16,37,38 Antibodies usually have high binding affinity and specificity, but their large size limits their tumor penetration. Several antibodies against PSMA have been applied for the diagnosis of prostate cancer.…”

Section: Discussionmentioning

confidence: 99%

Prostate-specific membrane antigen targeted protein contrast agents for molecular imaging of prostate cancer by MRI

Salarian

Xue

et al. 2016

Nanoscale

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Prostate-specific membrane antigen (PSMA) is one of the most specific cell surface markers for prostate cancer diagnosis and targeted treatment. However, achieving molecular imaging using non-invasive MRI with high resolution has yet to be achieved due to the lack of contrast agents with significantly improved relaxivity for sensitivity, targeting capabilities and metal selectivity. We have previously reported our creation of a novel class of protein Gd 3+ contrast agents, ProCA32, which displayed significantly improved relaxivity while exhibiting strong Gd 3+ binding selectivity over physiological metal ions. In this study, we report our effort in further developing biomarker-targeted protein MRI contrast agents for molecular imaging of PSMA. Among three PSMA targeted contrast agents engineered with addition of different molecular recognition sequences, ProCA32.PSMA exhibits a binding affinity of 1.1 ± 0.1 μM for PSMA while the metal binding affinity is maintained at 0.9 ± 0.1 × 10 −22 M. In addition, ProCA32.PSMA exhibits r 1 of 27.6 mM −1 s −1 and r 2 of 37.9 mM −1 s −1 per Gd (55.2 and 75.8 mM −1 s −1 per molecule r 1 and r 2 , respectively) at 1.4 T. At 7 T, ProCA32.PSMA also has r 2 of 94.0 mM −1 s −1 per Gd (188.0 mM −1 s −1 per molecule) and r 1 of 18.6 mM −1 s −1 per Gd (37.2 mM −1 s −1 per molecule). This contrast capability enables the first MRI enhancement dependent on PSMA expression levels in tumor bearing mice using both T 1 and T 2 -weighted MRI at 7 T. Further development of these PSMAtargeted contrast agents are expected to be used for the precision imaging of prostate cancer at an † Electronic supplementary information (ESI) available. See

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A strain-promoted alkyne–azide cycloaddition (SPAAC) reaction of a novel EpCAM aptamer–fluorescent conjugate for imaging of cancer cells

Cited by 21 publications

References 26 publications

Molecular Elucidation of Disease Biomarkers at the Interface of Chemistry and Biology

Molecular Elucidation of Disease Biomarkers at the Interface of Chemistry and Biology

Multiparametric Biomechanical and Biochemical Phenotypic Profiling of Single Cancer Cells Using an Elasticity Microcytometer

Prostate-specific membrane antigen targeted protein contrast agents for molecular imaging of prostate cancer by MRI

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