Abstract:
:IL-21 is a pleiotropic cytokine that plays a key role in modulating inflammatory responses, including the promotion of autoimmune diseases. Several groups have quantitated circulating levels of IL-21 in plasma and serum samples using various commercial ELISAs. We determined, however, that the most commonly used commercial assays in published literature were not specific or sensitive enough to detect levels of IL-21 in heparin plasma or serum from healthy human individuals. This finding prompted an effort to develop more specific and sensitive methods to quantitate IL-21 in complex biological matrices using proprietary anti-IL-21 antibodies with the Quanterix SiMoA platform and the Meso Scale Discovery (MSD) S-PLEX® format. Assays developed on both technology platforms were characterized in heparin plasma and serum using spike recoveries across a range of concentrations. Each method was able to detect sub-pg/mL levels of IL-21 (predicted Limit of Detection [LOD] of approximately 1.0 fg/mL for both the Quanterix SiMoA and MSD S-PLEX® platforms) which is 200-500 times lower than current commercial assays. Additionally we demonstrated that rheumatoid factor did not interfere with measuring IL-21 in the Quanterix SiMoA assay. Results obtained with the two new ultrasensitive assays showed a strong correlation (r = 0.9428; p < .0001). Additionally, IL-21 levels were significantly increased in samples from patients with Systemic Lupus Erythematosus (mean+/- SD: n = 14, 202.64 +/- 111.47 fg/mL, p = .0001 for Quanterix SiMoA and 275.4 +/- 174.66 fg/mL p = .0001 for MSD S-PLEX®) as well as in samples from patients with Sjögren's Syndrome (mean+/- SD: n = 11, 122.18 +/- 84.50 fg/mL, p = .0029 for Quanterix SiMoA and 183.64 +/- 153.00 fg/mL, p = .0082 for MSD S-PLEX®) when compared to healthy donors (mean+/- SD: n = 11, 38.1 +/- 27.8 fg/mL for Quanterix SiMoA and 58.1 +/- 30.7 fg/mL for MSD S-PLEX®). These ultrasensitive assays, for the first time, allow for the accurate quantitation of human IL-21 in heparin plasma and serum. In addition, these experiments also provide a direct comparison of the MSD S-PLEX® format and Quanterix SiMoA platform technologies, which may have broader implications to future application of these methods to evaluate low abundance proteins in complex biological matrices.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Poorbaugh J,Samanta T,Bright SW,Sissons SE,Chang CY,Oberoi P,MacDonald AJ,Martin AP,Cox KL,Benschop RJdoi
10.1016/j.jim.2018.12.005subject
Has Abstractpub_date
2019-03-01 00:00:00pages
9-16eissn
0022-1759issn
1872-7905pii
S0022-1759(18)30388-0journal_volume
466pub_type
杂志文章abstract::The identification of distinct T helper lymphocyte subsets (Th1/2) with polarised cytokine production has opened up new fields in immunobiology. Of the several alternative methods of monitoring cytokine production, flow cytometric analysis of intracellular staining has distinct advantages and pitfalls. It allows high ...
journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
doi:10.1016/s0022-1759(00)00230-1
更新日期:2000-09-21 00:00:00
abstract::Highly sensitive and easy detection method for Alzheimer's disease (AD) with a suitable biomarker is mandatory for preventing the factors resulting from AD. This research reports a modified ELISA with graphene for the detection of AD biomarker amyloid beta (Aβ) oligomer. Gold nanoparticle (AuNP) conjugated aptamer was...
journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(94)00310-s
更新日期:1995-03-27 00:00:00
abstract::An international collaborative study was conducted at ten sites to examine the performance of enzyme immunoassays (EIAs) for the quantitation of IgG1, IgG2, IgG3, IgG4 and total IgG anti-Haemophilus influenzae type b (Hib) capsular polysaccharide in human serum. All groups used the same reagents: microtiter plates coa...
journal_title:Journal of immunological methods
pub_type: 临床试验,杂志文章,多中心研究
doi:10.1016/0022-1759(92)90163-n
更新日期:1992-04-08 00:00:00
abstract::This paper describes a modification of the peroxidase technique by which immunophenotyping may be carried out on routinely air-dried blood and bone marrow (BM) smears. The method is simple and quick, requires no special equipment, can be performed on fresh or stored specimens and gives a standard of morphological deta...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90428-x
更新日期:1986-10-23 00:00:00
abstract::Two different methods have been used to study immune responses in the rat to Streptococcus pneumoniae type 3 and type 4 capsular polysaccharides (PPS). First, for simultaneous detection of the specificity and isotype of anti-PPS antibody-containing cells (ACC) in cryostat sections of lymphoid tissue, a double immunocy...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90314-6
更新日期:1991-12-15 00:00:00
abstract::Although enveloped retroviruses bud from the cell surface of T lymphocytes, they use the endocytic pathway and the internal membrane of multivesicular bodies for their assembly and release from macrophages and dendritic cells (DCs). Exosomes, physiological nanoparticles produced by hematopoietic cells, egress from thi...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2008.07.007
更新日期:2008-09-30 00:00:00
abstract::During a phase III pertussis vaccine trial, serum antibody responses were measured by two enzyme-linked immunosorbent assays (ELISA) for pertussis toxin and filamentous haemagglutinin. These were used both for studies of antibody levels after vaccination and for diagnostic purposes. Since the absorbance values obtaine...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(95)00067-k
更新日期:1995-06-28 00:00:00
abstract::The attachment of cells to plastic tissue culture plates is a widely used criterion of cell viability in microcytotoxicity assays. When cell suspensions of primary human colon carcinoma and melanoma cells which were of low initial viability (assessed by trypan blue exclusion) were allowed to adhere to tissue culture p...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(81)90039-9
更新日期:1981-01-01 00:00:00
abstract::A high-capacity manifold support with immobilized antigen was developed for the analysis of IgE-mediated immune reactivity in allergic subjects. Using this 96-pronged support, specific antibodies were trapped and detected from large sets of serum samples. We describe the binding of large amounts of antigen onto the ex...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(94)90218-6
更新日期:1994-01-12 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(77)90121-1
更新日期:1977-01-01 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(83)90222-3
更新日期:1983-07-29 00:00:00
abstract::The aim of this study was to compare the results of serological assays using pneumococcal proteins or polysaccharides for the detection of pneumococcal infection in childhood pneumonia. Serological assays measured IgG against eight pneumococcal proteins (Ply,CbpA,PspA1,PspA2,PcpA,PhtD,StkP-C,PcsB-N), C-polysaccharide ...
journal_title:Journal of immunological methods
pub_type: 临床试验,杂志文章,多中心研究
doi:10.1016/j.jim.2018.06.011
更新日期:2018-09-01 00:00:00
abstract::Thrombospondin (TSP), a platelet glycoprotein, was purified from platelet supernatants applied sequentially to Sepharose 4B and heparin-agarose affinity columns. This TSP inhibited alternative pathway activity in serum as assessed by lysis of rabbit erythrocytes and contained bound heparin, a substance which is known ...
journal_title:Journal of immunological methods
pub_type: 信件
doi:10.1016/s0022-1759(97)00006-9
更新日期:1997-03-28 00:00:00
abstract::A study was undertaken to establish conditions of growth to increase C3b receptor synthesis on a human B lymphoblastoid cell line (Raji) by use of cell differentiation effectors. It appears that whereas two polar compounds HMBA (2mM) and Me2SO(2%) have no or little effect, 5 BrdU (30microM) and db cAMP (5 x10(-4) M) a...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(81)90135-6
更新日期:1981-01-01 00:00:00
abstract::A method for bleeding rats from the tail vein was developed. Sufficient vasodilation of the tail was accomplished by the use of a specially designed warming device. With the help of a physical restrainer, up to 5 ml of blood can be collected from the tail vein and no anaesthesia is required. This bleeding procedure is...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90148-x
更新日期:1982-09-17 00:00:00
abstract::After centrifugation of heparinised whole blood on Hypaque-Ficoll medium (s.g. 1.114, Ferrante and Thong, 1980) the specific gravities were measured of the layers of medium above and below the band of polymorphonuclear leukocytes (PMN), as well as of the PMN and sedimented erythrocytes. The erythrocytes and PMN were a...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90363-2
更新日期:1987-01-26 00:00:00
abstract::A sensitive, specific immunoassay for detection of hepatitis B surface antigen (HBsAg) is described. The assay combined enzyme-linked immunosorbent assay and solid-phase radioimmunoassay and is termed enzyme potentiated radioimmunoassay (EPRIA). HBsAg was quantitated by enzymatic conversion of L[14C]glutamic acid to 1...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(81)90115-0
更新日期:1981-01-01 00:00:00
abstract::An antibody binding-inhibition test is described, which allows the detection of P. falciparum in red blood cells (RBC) infected in vitro, using a crossreacting, monoclonal anti-P. berghei antibody and P. berghei coated microtiter plates. Experiments carried out to determine the coating efficiency of various P. berghei...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90231-5
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journal_title:Journal of immunological methods
pub_type: 杂志文章
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doi:10.1016/s0022-1759(96)00147-0
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90516-3
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abstract::A new quantitative assay for migration inhibitory factor (MIF) employs 3H-labelled cultured mouse or human lymphoid cells migrating from capillary tubes. Capillaries filled with labelled cells are placed in liquid scintillation counting vials, along with the MIF-containing sample and are removed at the end of a five-h...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(78)90258-2
更新日期:1978-01-01 00:00:00
abstract::An immunomagnetic technique to detect and identify Salmonella serogroup C1 has been developed. Monoclonal antibodies (MAbs) specific for the O antigen 6,7 of Salmonella lipopolysaccharide (LPS) coupled to magnetic beads were used to isolate the salmonellae. Captured bacteria were easily identified by acridine orange s...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90387-u
更新日期:1991-03-01 00:00:00
abstract::Foot-and-mouth disease virus (FMDV) is an important veterinary pathogen which can cause widespread epidemics. Due to the high antigenic variability of FMDV, it is important to undertake mutation analysis under immunological pressure. To study the bovine antibody response at a molecular level, phage display technology ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2004.01.002
更新日期:2004-03-01 00:00:00
abstract::Dengue fever is caused by the dengue virus (DENV), and DENV1 is the prevalent epidemic serotype in south China. A new lateral flow assay (LFA) based on a near-infrared (NIR) fluorescent dye was developed to detect anti-DENV1 IgG antibodies. DyLight-800 was used as the marker conjugated to goat anti-human IgG antibodie...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2018.02.005
更新日期:2018-05-01 00:00:00
abstract::Methods for the dissociation of immunoglobulin-Brucella complexes (IgM-rich) were evaluated. The antigen-antibody complex was separated from serum by centrifugation and dissociated with either H2O, 2.0 M pH 2.0 glycine buffer, or various concentrations of KSCN. Optimal antibody recoveries for KSCN were obtained using ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(79)90152-2
更新日期:1979-01-01 00:00:00
abstract::The sensitivity of any immunoassay is a complex function of the underlying physico-chemical basis of the technique and the size and source of 'experimental' errors. Analysis of this relationship emphasizes the distinction between competitive labelled analyte methods, e.g., RIA, and the non-competitive labelled antibod...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90338-8
更新日期:1986-02-27 00:00:00
abstract::Hybridoma cells producing monoclonal antibodies against tumor-shed antigens were generated by fusing mouse myeloma cells with spleen cells from immunosuppressed mice bearing human melanoma xenografts. Thirty-eight fusion experiments were performed at different stages of tumor growth. Hybridomas producing anti-melanoma...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(83)90074-1
更新日期:1983-02-25 00:00:00
abstract::Affinity chromatography and subsequent ion-exchange chromatography of pooled anti-benzylpenicilloyl (anti-BPO) hyperimmune sera separated 5 different anti-BPO IgG fractions as described in the preceding paper. These fractions were tested for activities in passive hemagglutination (PHA), passive immune hemolysis (PIH),...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(78)90163-1
更新日期:1978-01-01 00:00:00