scholarly journals Analytical Currents: Nanotube electrodes measure DNA conductivity | Flow cytometry: the eyes have it | Functional analysis of membrane proteins with nanodiscs and SAMDI MS | Immunomagnetic diffractometry for biomarker detection | Pushing molecules back and forth through a nanopore | The mystery of the pink paintings| Shared Instrumentation Grant program to accept proposals

2008 ◽  
Vol 80 (3) ◽  
pp. 513-515
Keyword(s):  
Flow Cytometry ◽  
Functional Analysis ◽  
Membrane Proteins ◽  
Biomarker Detection ◽  
Grant Program ◽  
Dna Conductivity

scholarly journals Co-Translational Insertion of Aquaporins into Liposome for Functional Analysis via an E. coli Based Cell-Free Protein Synthesis System

Cells ◽  
2019 ◽  
Vol 8 (11) ◽  
pp. 1325 ◽  
Author(s):  
Ke Yue ◽  
Tran Nam Trung ◽  
Yiyong Zhu ◽  
Ralf Kaldenhoff ◽  
Lei Kai
Keyword(s):  
Functional Analysis ◽  
Membrane Proteins ◽  
Fluorescent Protein ◽  
Water Channel ◽  
New Approach ◽  
E Coli ◽  
Straightforward Method ◽  
Lipid Environment ◽  
Green Fluorescent ◽  
Eukaryotic Organisms

Aquaporins are important and well-studied water channel membrane proteins. However, being membrane proteins, sample preparation for functional analysis is tedious and time-consuming. In this paper, we report a new approach for the co-translational insertion of two aquaporins from Escherichia coli and Nicotiana tabacum using the CFPS system. This was done in the presence of liposomes with a modified procedure to form homogenous proteo-liposomes suitable for functional analysis of water permeability using stopped-flow spectrophotometry. Two model aquaporins, AqpZ and NtPIP2;1, were successfully incorporated into the liposome in their active forms. Shifted green fluorescent protein was fused to the C-terminal part of AqpZ to monitor its insertion and status in the lipid environment. This new fast approach offers a fast and straightforward method for the functional analysis of aquaporins in both prokaryotic and eukaryotic organisms.

scholarly journals Culture, expansion, and flow-cytometry-based functional analysis of pteropid bat MR1-restricted unconventional T cells

2021 ◽  
Vol 2 (2) ◽  
pp. 100487
Author(s):  
Wan Rong Sia ◽  
Ying Ying Hey ◽  
Randy Foo ◽  
Lin-Fa Wang ◽  
Edwin Leeansyah
Keyword(s):  
Flow Cytometry ◽  
Functional Analysis ◽  
T Cells

scholarly journals 883 An anti-carcinoma monoclonal antibody (mAb) NEO-201 can also target human acute myeloid leukemia (AML) cell lines in vitro

2021 ◽  
Vol 9 (Suppl 3) ◽  
pp. A925-A925
Author(s):  
Alessandra Romano ◽  
Nunziatina Parrinello ◽  
Sara Marino ◽  
Enrico La Spina ◽  
Massimo Fantini ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Functional Analysis ◽  
Epithelial Cells ◽  
Cell Line ◽  
Cell Lines ◽  
Lymphoblastic Leukemia ◽  
Phase 1 ◽  
Adcc Activity ◽  
Phenotypic Analysis

BackgroundNEO-201 is an IgG1 mAb targeting variants of CEACAM5/6 and has demonstrated tumor sensitivity and specificity in epithelial cells. Functional analysis has revealed that NEO-201 can engage innate immune effector mechanisms including ADCC and CDC to directly kill tumor cells expressing its target. A recent Phase 1 clinical trial at the NCI has determined both safety and recommended Phase 2 dosing. We have also seen the expression of the NEO-201 target on hematologic cells, specifically Tregs and neutrophils. Due to epitope being expressed both on malignant epithelial cells as well as several hematologic cells, we designed this study to explore the reactivity of NEO-201 against hematological neoplastic cells in vitro.MethodsPhenotypic analysis was conducted by flow cytometry. Cell lines used were six AML (HL60, U937, MOLM13, AML2, IMS-M2 and OCL-AML3), two multiple myelomas (MM) (OPM2, MM1.S), two acute lymphoblastic leukemia (ALL) (SUP-B15, RPMI8402) and four mantle cell lymphoma (MCL) (Jeko-1, Z138, JVM2 and JVM13). Markers used for flow cytometry analysis were CD15, CD45, CD38, CD138, CD14, CD19 and NEO-201. Functional analysis was performed by evaluating the ability of NEO-201 to mediate ADCC activity against AML cell lines using human NK cells as effector cells.Results5 of 6 AML cell lines tested bind to NEO-201 and the% of positive cells were 47%, 99.5%,100%,100% and 97.8% for HL60, U937, MOLM13, AML3 and IMS-M2, respectively. The% of positive cells in the two MM cell line were 99% and 18% for OPM2 and MM1.S, respectively. NEO-201 binding was not detected in the two ALL and the four MCL cell lines tested. Functional analysis has demonstrated that NEO-201 can mediate ADCC activity against the AML cell line (HL60) tested.ConclusionsThis study demonstrates that NEO-201 mAb’s target is expressed in most of the AML cell lines tested in vitro. In addition, we have shown it can mediate ADCC activity against HL60 cells (AML). Together, these findings provide a rationale for further investigation of the role of NEO-201 in AML as well as MM, further exploring patient PBMCs and bone marrow samples.

scholarly journals Functional Analysis of Anti-cytokine Autoantibodies Using Flow Cytometry

2019 ◽  
Vol 10 ◽  
Author(s):  
Patricia A. Merkel ◽  
Terri Lebo ◽  
Vijaya Knight
Keyword(s):  
Flow Cytometry ◽  
Functional Analysis

scholarly journals An Innovative Method to Identify Autoantigens Expressed on the Endothelial Cell Surface: Serological Identification System for Autoantigens Using a Retroviral Vector and Flow Cytometry (SARF)

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Tsuyoshi Shirai ◽  
Hiroshi Fujii ◽  
Masao Ono ◽  
Ryu Watanabe ◽  
Tomonori Ishii ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Endothelial Cell ◽  
Membrane Proteins ◽  
Cell Surface ◽  
Transmembrane Protein ◽  
Retroviral Vector ◽  
Vascular Lesions ◽  
Intercellular Adhesion Molecule ◽  
Identification System ◽  
Serological Identification

Autoantibodies against integral membrane proteins are usually pathogenic. Although anti-endothelial cell antibodies (AECAs) are considered to be critical, especially for vascular lesions in collagen diseases, most molecules identified as autoantigens for AECAs are localized within the cell and not expressed on the cell surface. For identification of autoantigens, proteomics and expression library analyses have been performed for many years with some success. To specifically target cell-surface molecules in identification of autoantigens, we constructed a serological identification system for autoantigens using a retroviral vector and flow cytometry (SARF). Here, we present an overview of recent research in AECAs and their target molecules and discuss the principle and the application of SARF. Using SARF, we successfully identified three different membrane proteins: fibronectin leucine-rich transmembrane protein 2 (FLRT2) from patients with systemic lupus erythematosus (SLE), intercellular adhesion molecule 1 (ICAM-1) from a patient with rheumatoid arthritis, and Pk (Gb3/CD77) from an SLE patient with hemolytic anemia, as targets for AECAs. SARF is useful for specific identification of autoantigens expressed on the cell surface, and identification of such interactions of the cell-surface autoantigens and pathogenic autoantibodies may enable the development of more specific intervention strategies in autoimmune diseases.

scholarly journals Advances in Structural and Functional Analysis of Membrane Proteins by Electron Crystallography

Structure ◽  
2011 ◽  
Vol 19 (10) ◽  
pp. 1381-1393 ◽  
Author(s):  
Goragot Wisedchaisri ◽  
Steve L. Reichow ◽  
Tamir Gonen
Keyword(s):  
Functional Analysis ◽  
Membrane Proteins ◽  
Electron Crystallography

scholarly journals NK Cell Isolation from Liver Biopsies: Phenotypic and Functional Analysis of Low Cell Numbers by Flow Cytometry

2013 ◽  
Vol 4 ◽  
Author(s):  
Ning Li ◽  
Gisella L. Puga Yung ◽  
Amandine Pradier ◽  
Christian Toso ◽  
Emiliano Giostra ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Functional Analysis ◽  
Nk Cell ◽  
Cell Isolation ◽  
Cell Numbers ◽  
Liver Biopsies

scholarly journals Preparation, identification, and functional analysis of monoclonal antibodies against atypical porcine pestivirus NS3 protein

2020 ◽  
Vol 32 (5) ◽  
pp. 695-699
Author(s):  
Miaomiao Yan ◽  
Jingling Huang ◽  
Jianing Chen ◽  
Wenting Yang ◽  
Guangliang Liu
Keyword(s):  
Flow Cytometry ◽  
Functional Analysis ◽  
Monoclonal Antibodies ◽  
Western Blot ◽  
Light Chain ◽  
Immunofluorescence Assay ◽  
Linear Epitope ◽  
Kappa Light Chain ◽  
Conformational Epitopes ◽  
Ns3 Protein

Atypical porcine pestivirus (APPV) had been detected in many countries. However, to date, a commercial detection kit is not available because of a lack of specific monoclonal antibodies (mAbs) to APPV. We generated 7 mAbs targeting the NS3 protein of APPV. Isotyping results indicated that all of these mAbs are IgG1 with a kappa light chain. We analyzed the epitopes recognized by mAbs 2B6, 6G11, 8D1, 8D3, and 8F12, which recognized the same linear epitope (GRIKSAYSDE); the 6H3 and 7E10 mAbs recognized 2 different conformational epitopes. Applications of these antibodies were verified by ELISA, western blot, indirect immunofluorescence assay, and flow cytometry. The antibodies were functionally workable for these immunoassays except for 8F12, which could not be used in flow cytometry.

scholarly journals Multi-Dimensional T-Cell Functional Analysis by Spectral Flow Cytometry as a Tool to Dissect Endotypes in Allergic Disease.

2019 ◽  
Vol 143 (2) ◽  
pp. AB184
Author(s):  
Alberta GA. Paul ◽  
Jill Glesner ◽  
Josephine Lannigan ◽  
Lyndsey M. Muehling ◽  
Jacob D. Eccles ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Functional Analysis ◽  
T Cell ◽  
Allergic Disease ◽  
Spectral Flow

Generation of membrane proteins in polymer-based lipoparticles as flow cytometry antigens

2018 ◽  
Vol 109 ◽  
pp. 483-488 ◽  
Author(s):  
V. Mitch Luna ◽  
Mukta Vazir ◽  
Amit Vaish ◽  
Su Chong ◽  
Irwin Chen ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Membrane Proteins
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