Application of immuno-PCR for the detection of early stage cancer

Khan, Amjad Hayat; Sadroddiny, Esmaeil

doi:10.1016/j.mcp.2016.01.010

Cited by 28 publications

(11 citation statements)

References 52 publications

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“…The spPLA for survivin resulted in a limit of detection of 1.45 pg/mL, which is about 23-times better than the limit of detection of the survivin ELISA [ 19 ]. This constitutes a major improvement for survivin detection and is in accordance with data from the literature for immuno-PCR assays showing an improved limit of detection for different protein targets [ 30 , 48 , 49 ]. Initial experiments of a homogenous PLA failed, most likely due to inhibiting elements in urine [ 6 ].…”

Section: Discussionsupporting

confidence: 87%

Exploring solid-phase proximity ligation assay for survivin detection in urine

et al. 2022

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Urine-based biomarkers are a rational and promising approach for the detection of bladder cancer due to the proximity of urine to the location of the tumor site and the non-invasive nature of its sampling. A well-known and highly investigated biomarker for bladder cancer is survivin. For detection of very small amounts of urinary survivin protein a highly sensitive assay was developed. The assay is based on the immuno-PCR technology, more precisely a solid-phase proximity ligation assay (spPLA). The limit of detection for the survivin spPLA was 1.45 pg/mL, resulting in an improvement of the limit of detection by a factor of approximately 23 compared to the previously in-house developed survivin ELISA. A key step in development was the initial isolation of survivin by a molecular fishing rod based on magnetic beads. Interfering matrix compounds pose a special challenge for further analytical application, but can be overcome by this isolation step. The assay is designed to work with only 500 μL of voided urine. The survivin spPLA showed a sensitivity of 30% and specificity of 89% for bladder cancer detection in this study of 110 bladder cancer cases and 133 clinical controls. Moreover, the results demonstrated again that survivin is a useful complementary marker in combination with UBC® Rapid by increasing the overall sensitivity to 70% with a specificity of 86%. Although the performance for detection of bladder cancer was rather low, the herein developed assay might serve as a new tool for survivin biomarker research in diverse human fluids, even if the biological matrix is complex or survivin is only present in small amounts.

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Section: Discussionsupporting

confidence: 87%

Exploring solid-phase proximity ligation assay for survivin detection in urine

et al. 2022

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“…Detailed protocol for immunoPCR assay development for IL-6 in plasma was described by [ 161 ]. Applications of immuno-PCR for the detection of early stage cancer were currently reviewed in [ 162 , 163 ].…”

Section: Cytokine Detection Techniquesmentioning

confidence: 99%

Advances in Proteomic Techniques for Cytokine Analysis: Focus on Melanoma Research

Skalníková

Čížková

Cervenka

et al. 2017

IJMS

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Melanoma is a skin cancer with permanently increasing incidence and resistance to therapies in advanced stages. Reports of spontaneous regression and tumour infiltration with T-lymphocytes makes melanoma candidate for immunotherapies. Cytokines are key factors regulating immune response and intercellular communication in tumour microenvironment. Cytokines may be used in therapy of melanoma to modulate immune response. Cytokines also possess diagnostic and prognostic potential and cytokine production may reflect effects of immunotherapies. The purpose of this review is to give an overview of recent advances in proteomic techniques for the detection and quantification of cytokines in melanoma research. Approaches covered span from mass spectrometry to immunoassays for single molecule detection (ELISA, western blot), multiplex assays (chemiluminescent, bead-based (Luminex) and planar antibody arrays), ultrasensitive techniques (Singulex, Simoa, immuno-PCR, proximity ligation/extension assay, immunomagnetic reduction assay), to analyses of single cells producing cytokines (ELISpot, flow cytometry, mass cytometry and emerging techniques for single cell secretomics). Although this review is focused mainly on cancer and particularly melanoma, the discussed techniques are in general applicable to broad research field of biology and medicine, including stem cells, development, aging, immunology and intercellular communication.

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“…Moreover, newly-developed ultrasensitive protein assays like immuno-PCR could be seen as an incremental step in the field of immunological research and clinical diagnostics. The techniques are combined for nucleic acid amplification with an antibody-based assay which can dramatically increase the sensitivity of conventional immunoassays [25][26][27][28]. These methods can quantify up to a few dozen endogenous proteins recognized by highly specific antibodies and have enabled a lot many exciting research avenues.…”

Section: Discussionmentioning

confidence: 99%

Laser Capture Microdissection Coupled Capillary Immunoassay to Study the Expression of PCK-2 on Spatially-Resolved Islets of Rat Langerhans

et al. 2021

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The platform for precise proteomic profiling of targeted cell populations from heterogeneous tissue sections is developed. We demonstrate a seamless and systematic integration of LCM with an automated cap-IA for the handling of a very small-sized dissected tissues section from the kidney, liver and pancreatic Langerhans islet of rats. Our analysis reveals that the lowest LCM section area ≥ 0.125 mm2 with 10 µm thickness can be optimized for the detection of proteins through LCM-cap-IA integration. We detect signals ranging from a highly-abundant protein, β-actin, to a low-abundance protein, LC-3AB, using 0.125 mm2 LCM section from rat kidney, but, so far, a relatively large section is required for good quality of results. This integration is applicable for a highly-sensitive and accurate assessment of microdissected tissue sections to decipher hidden proteomic information of pure targeted cells. To validate this integration, PCK2 protein expression is studied within Langerhans islets of normal and diabetic rats. Our results show significant overexpression of PCK2 in Langerhans islets of rats with long-term diabetes.

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Application of immuno-PCR for the detection of early stage cancer

Cited by 28 publications

References 52 publications

Exploring solid-phase proximity ligation assay for survivin detection in urine

Exploring solid-phase proximity ligation assay for survivin detection in urine

Advances in Proteomic Techniques for Cytokine Analysis: Focus on Melanoma Research

Laser Capture Microdissection Coupled Capillary Immunoassay to Study the Expression of PCK-2 on Spatially-Resolved Islets of Rat Langerhans

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