Profiling CD antigens on leukaemias with an antibody microarray

Barber, Nicole; Gez, Swetlana; Belov, Larissa; Mulligan, Stephen P.; Woolfson, Adrian; Christopherson, Richard I.

doi:10.1016/j.febslet.2009.03.018

Cited by 23 publications

(14 citation statements)

References 45 publications

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“…Proof of principle of this technology in leukemia diagnosis has already been reported [2]. Another approach uses antibody microarrays based on a solid phase cell-capture technique [3], [4]. Belov L et al reported the analysis of the simultaneous expression of eighty-two antigens in 733 patients with a variety of leukemias and lymphomas, achieving a high degree of diagnostic consensus between established criteria and the results of an antibody microarray (93.9% for peripheral blood and 97.6% for bone marrow aspirates) [3].…”

Section: Introductionmentioning

confidence: 99%

Using Digital RNA Counting and Flow Cytometry to Compare mRNA with Protein Expression in Acute Leukemias

et al. 2012

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BackgroundThe diagnosis of malignant hematologic diseases has become increasingly complex during the last decade. It is based on the interpretation of results from different laboratory analyses, which range from microscopy to gene expression profiling. Recently, a method for the analysis of RNA phenotypes has been developed, the nCounter technology (Nanostring® Technologies), which allows for simultaneous quantification of hundreds of RNA molecules in biological samples. We evaluated this technique in a Swiss multi-center study on eighty-six samples from acute leukemia patients.MethodsmRNA and protein profiles were established for normal peripheral blood and bone marrow samples. Signal intensities of the various tested antigens with surface expression were similar to those found in previously performed Affymetrix microarray analyses. Acute leukemia samples were analyzed for a set of twenty-two validated antigens and the Pearson Correlation Coefficient for nCounter and flow cytometry results was calculated.ResultsHighly significant values between 0.40 and 0.97 were found for the twenty-two antigens tested. A second correlation analysis performed on a per sample basis resulted in concordant results between flow cytometry and nCounter in 44–100% of the antigens tested (mean = 76%), depending on the number of blasts present in a sample, the homogeneity of the blast population, and the type of leukemia (AML or ALL).ConclusionsThe nCounter technology allows for fast and easy depiction of a mRNA profile from hematologic samples. This technology has the potential to become a valuable tool for the diagnosis of acute leukemias, in addition to multi-color flow cytometry.

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Section: Introductionmentioning

confidence: 99%

Using Digital RNA Counting and Flow Cytometry to Compare mRNA with Protein Expression in Acute Leukemias

et al. 2012

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“…Another advantage of using array separation devices is that the volume of the array fluidic chamber is typically well known, allowing for absolute cell counting and eliminating the need for counting beads or an additional counting step – a large bottle neck in separations today. Affinity arrays have been extensively developed for characterization of blood cells and other cells based on antibody–antigen capture (Barber et al, 2009, Kaufman et al, 2010, Kohnke et al, 2009, Rahman et al, 2012, Zhou et al, 2010). …”

Section: Overview Of Cell Separation Methodsmentioning

confidence: 99%

Fundamentals and application of magnetic particles in cell isolation and enrichment: a review

2014

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Magnetic sorting using magnetic beads has become a routine methodology for the separation of key cell populations from biological suspensions. Due to the inherent ability of magnets to provide forces at a distance, magnetic cell manipulation is now a standardized process step in numerous processes in tissue engineering, medicine, and in fundamental biological research. Herein we review the current status of magnetic particles to enable isolation and separation of cells, with a strong focus on the fundamental governing physical phenomena, properties and syntheses of magnetic particles and on current applications of magnet-based cell separation in laboratory and clinical settings. We highlight the contribution of cell separation to biomedical research and medicine and detail modern cell separation methods (both magnetic and non-magnetic). In addition to a review of the current state-of-the-art in magnet-based cell sorting, we discuss current challenges and available opportunities for further research, development and commercialization of magnetic particle-based cell separation systems.

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“…Complex CD antigen phenotyping is achievable with DotScan TM [4,5], by printing cell surface antigen-specific antibodies onto the array, then incubating the slide in cell suspension, and finally visualizing bound cells. The binding pattern indicates which CD antigens are expressed on the cell membrane.…”

Section: Determination Of Cluster Of Differentiation (Cd) Antigensmentioning

confidence: 99%

Life on a microarray: assessing live cell functions in a microarray format

Papp

Szittner

Prechl

2012

Cell. Mol. Life Sci.

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Microarray technology outgrew the detection of simple intermolecular interactions, as incubation of slides with living cells opened new vistas. Cell-based array technology permits simultaneous detection of several different cell surface molecules, allowing the complex characterization of cells with an amount of information that is hardly assessed by any other technique. Furthermore, binding of cells to printed antibodies or ligands may induce their activation, and consequently the outcome of these interactions, such as phosphorylation, gene expression, secretion of various products; differentiation, proliferation and apoptosis of the cells are also measurable on arrays. Moreover, since cells can be transfected with printed vectors, over- or under-expression of selected genes is also achievable simultaneously, creating a nice tool for assessing the function of a given gene. The enormously high-throughput cell-based microarray technology enables testing the effect of external stimuli on a scale that was earlier unthinkable. This review summarizes the possible applications of cell-based arrays.

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Profiling CD antigens on leukaemias with an antibody microarray

Cited by 23 publications

References 45 publications

Using Digital RNA Counting and Flow Cytometry to Compare mRNA with Protein Expression in Acute Leukemias

Using Digital RNA Counting and Flow Cytometry to Compare mRNA with Protein Expression in Acute Leukemias

Fundamentals and application of magnetic particles in cell isolation and enrichment: a review

Life on a microarray: assessing live cell functions in a microarray format

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