Immunoenzymatic Quantitative Analysis of Antigens Expressed on the Cell Surface (Cell-ELISA)

Lourenço, Elaine; Roque-Barreira, Maria-Cristina

doi:10.1007/978-1-59745-324-0_29

Cited by 18 publications

(12 citation statements)

References 16 publications

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“…The potential applications of cell‐based enzyme‐linked immunosorbent assays (cell‐ELISA) go far beyond hybridoma screening . Only recently, a neuroblastoma cell line‐based cell‐ELISA employed for screening for anti‐neuronal antibodies has been reported .…”

Section: Laboratory Tests For Nmo‐igg/aqp4‐abmentioning

confidence: 99%

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Aquaporin‐4 Antibodies (NMO‐IgG) as a Serological Marker of Neuromyelitis Optica: A Critical Review of the Literature

2013

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Antibodies to aquaporin-4 (called NMO-IgG or AQP4-Ab) constitute a sensitive and highly specific serum marker of neuromyelitis optica (NMO) that can facilitate the differential diagnosis of NMO and classic multiple sclerosis. NMO-IgG/AQP4-Ab seropositive status has also important prognostic and therapeutic implications in patients with isolated longitudinally extensive myelitis (LETM) or optic neuritis (ON). In this article, we comprehensively review and critically appraise the existing literature on NMO-IgG/AQP4-Ab testing. All available immunoassays-including tissue-based (IHC), cell-based (ICC, FACS) and protein-based (RIPA, FIPA, ELISA, Western blotting) assays-and their differential advantages and disadvantages are discussed. Estimates for sensitivity, specificity, and positive and negative likelihood ratios are calculated for all published studies and accuracies of the various immunoassay techniques compared. Subgroup analyses are provided for NMO, LETM and ON, for relapsing vs. monophasic disease, and for various control groups (eg, MS vs. other controls). Numerous aspects of NMO-IgG/AQP4-Ab testing relevant for clinicians (eg, impact of antibody titers and longitudinal testing, indications for repeat testing, relevance of CSF testing and subclass analysis, NMO-IgG/AQP4-Ab in patients with rheumatic diseases) as well as technical aspects (eg, AQP4-M1 vs. AQP4-M23-based assays, intact AQP4 vs. peptide substrates, effect of storage conditions and freeze/thaw cycles) and pitfalls are discussed. Finally, recommendations for the clinical application of NMO-IgG/ AQP4-Ab serology are given.

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Section: Laboratory Tests For Nmo‐igg/aqp4‐abmentioning

confidence: 99%

“…However, controlling for non‐specific binding is more difficult in these types of assays than in conventional ICC qualitatively analyzed by a human rater. To date, no cell‐ELISAs for the detection of NMO‐IgG/AQP4‐Ab have been published .…”

Section: Laboratory Tests For Nmo‐igg/aqp4‐abmentioning

confidence: 99%

Aquaporin‐4 Antibodies (NMO‐IgG) as a Serological Marker of Neuromyelitis Optica: A Critical Review of the Literature

2013

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“…Expression of FLAG-tagged LHRs in intact or permeabilized cells was measured by ELISA [an established method for measurement of GPCR expression (11)(12)(13)(14)], as a ligand-independent method for determining total and cell surface receptor expression, respectively. In each case, expression of the mutant LHRs was assessed and compared with that measured for the wild-type receptor in the same experiment.…”

Section: Quantification Of Receptor Expression By Elisamentioning

confidence: 99%

Loss-of-Function Mutations in the Human Luteinizing Hormone Receptor Predominantly Cause Intracellular Retention

et al. 2016

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Mutations in G protein-coupled receptors (GPCRs) have been identified for many endocrine hormone signaling deficiencies. Inactivating mutations can impair ligand binding, receptor activation/ coupling to signaling pathways, or can cause receptor misfolding and consequent impaired expression at the cell membrane. Here we examine the cell surface expression, ligand binding, and signaling of a range of mutant human luteinizing hormone receptors (LHRs) identified as causing reproductive dysfunction in human patients. The data obtained reveal how mutations in GPCRs can have diverse and severely deleterious effects on receptor function. Furthermore, it was found that impaired functionality of the majority of the mutant LHRs was due to reduced expression at the cell surface (14/20) while only two mutations caused impaired binding affinity and two impaired in signaling. An additional two mutations were found to cause no impairment of receptor function. These data demonstrate that the majority of LHR mutations lead to intracellular retention and highlight the potential for novel pharmacological chaperone therapeutics that can "rescue" expression/function of retained mutant GPCRs. (Endocrinology 157: 4364 -4377, 2016) T he hypothalamic-pituitary-gonadal axis governs the endocrine control of reproduction. GnRH released from the hypothalamus binds the GnRH receptor on gonadotrope cells in the anterior pituitary, stimulating secretion of the gonadotropins, LH and FSH. LH and FSH enter the circulation and activate their cognate receptors LHR/LHCGR and FSHR in the gonads, to stimulate gametogenesis and the production/secretion of the sex steroid hormones. Perturbation of the hypothalamicpituitary-gonadal axis is associated with reproductive phenotypes, such as Leydig cell hypoplasia (LCH) in males (manifesting as a continuum of disorders from micropenis and hypospadias [Class II LCH], through to pseudohermaphroditism [Class I LCH]), and amenorrhea, and ovarian cysts in females (1). LCH is a 46,XY disorder, characterized by high circulating levels of LH with impaired male gonadal development. In some cases, LCH can be attributed to inactivating mutations in the LHR gene. In 46,XX patients inactivating LHR mutations can result in infertility, oligo-/amenorrhea, and/or empty follicle syn-

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“…We therefore determined the steady-state levels on the cell surface of all the mutant Env glycoproteins and compared them with those of wild-type FIV Env using cellular enzyme-linked immunosorbent assay (cellular ELISA). This technique is highly sensitive and specific for the quantitative analysis of molecules expressed on the cell surface (Jó zefowski et al, 2011;Lourenc -o and Roque-Barreira, 2010). 293T cells expressing the wild-type or mutant Env glycoproteins were incubated first with the anti-SU monoclonal antibody (MAb) and then with a horseradish peroxidase (HRP)-coupled secondary antibody as described in ''Materials and methods''.…”

Section: Cell Surface Expression Of Fiv Env Mutantsmentioning

confidence: 99%

Palmitoylation of the feline immunodeficiency virus envelope glycoprotein and its effect on fusion activity and envelope incorporation into virions

2012

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Immunoenzymatic Quantitative Analysis of Antigens Expressed on the Cell Surface (Cell-ELISA)

Cited by 18 publications

References 16 publications

Aquaporin‐4 Antibodies (NMO‐IgG) as a Serological Marker of Neuromyelitis Optica: A Critical Review of the Literature

Aquaporin‐4 Antibodies (NMO‐IgG) as a Serological Marker of Neuromyelitis Optica: A Critical Review of the Literature

Loss-of-Function Mutations in the Human Luteinizing Hormone Receptor Predominantly Cause Intracellular Retention

Palmitoylation of the feline immunodeficiency virus envelope glycoprotein and its effect on fusion activity and envelope incorporation into virions

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