样式: 排序: IF: - GO 导出 标记为已读
-
Development of a defibrinated human blood hemolysis assay for rapid testing of hemolytic activity compared to computational prediction J. Immunol. Methods (IF 2.2) Pub Date : 2024-04-09 Ashley M. Carpenter, Monique L. van Hoek
-
Humoral and cellular response of two different vaccines against SARS-CoV-2 in a group of healthcare workers: Comment J. Immunol. Methods (IF 2.2) Pub Date : 2024-04-07 Hinpetch Daungsupawong, Viroj Wiwanitkit
-
Protein G affinity chromatography is an underrated but very potent purification method for a broad range of species-independent and tag-less Fab-fragments J. Immunol. Methods (IF 2.2) Pub Date : 2024-04-04 Daniel Stern, Paulin Dettmann, Brigitte G. Dorner, Hans Werner Mages
Because of their superior properties for certain biological applications small antibody derivatives like fragment of antigen binding (Fab) have found widespread use in basic research and as therapeutics. However, generation of Fab-fragments is still a rather complex matter, reflected by the fact that a variety of methods and purification techniques are necessary for the production of all the different
-
Validation of dot blot immunoassay for measurement of complement opsonization of nanoparticles J. Immunol. Methods (IF 2.2) Pub Date : 2024-04-02 Yue Li, Andrew Monte, Layne Dylla, S. Moein Moghimi, Dmitri Simberg
-
Magnetic CAR T cell purification using an anti-G4S linker antibody J. Immunol. Methods (IF 2.2) Pub Date : 2024-04-02 Dennis Christoph Harrer, Sin-Syue Li, Marcell Kaljanac, Valerie Bezler, Markus Barden, Hong Pan, Wolfgang Herr, Hinrich Abken
Chimeric antigen receptor (CAR) redirected T cells are successfully employed in the combat against several hematological malignancies, however, are often compromised by low transduction rates making refinement of the CAR T cell products necessary. Here, we report a broadly applicable enrichment protocol relying on marking CAR T cells with an anti-glycine-serine (G4S) linker antibody followed by magnetic
-
Simultaneous determination of inflammatory factors SAA and LTF based on stable element labeling and inductively coupled plasma mass spectrometry to aid in the diagnosis of infection J. Immunol. Methods (IF 2.2) Pub Date : 2024-04-02 Hairong Tang, Gongwei Sun, Ying Xu, Shasha Men, Wencan Jiang, Chengbin Wang
The clinical value of Serum amyloid A (SAA) and Lactoferrin (LTF) has received significant attention, but their detection methods are inadequate, which limits their application. This study aims to develop a dual detection method based on stable element labeling strategies and inductively coupled plasma mass spectrometry (ICP-MS) for SAA/LTF and to assess whether it can be widely used in clinical practice
-
Humoral and cellular response of two different vaccines against SARS-CoV-2 in a group of healthcare workers: An observational study J. Immunol. Methods (IF 2.2) Pub Date : 2024-03-14 Nejla Stambouli, Khadija Bahrini, Chihebeddine Romdhani, Aicha Rebai, Sana Boughariou, Mohamed Zakraoui, Bilel Arfaoui, Sameh Seyli, Yasmine Boukhalfa, Riadh Battikh, Mohamed Ben Moussa, Iheb Labbene, Mustpha Ferjani, Hedi Gharssallah
On March 13, 2021, Tunisia started a widespread immunization program against SARS-CoV-2 utilizing different vaccinations that had been given emergency approval. Herein, we followed prospectively a cohort of participant who received COVID-19 vaccine (Pfizer BioNTech and Sputnik-Gameleya V). The goal of this follow-up was to define the humoral and cellular immunological profile after immunization by
-
An optimized non-T cell transfection system based on HEK293FT cells for CD3ζ phosphorylation and ubiquitination J. Immunol. Methods (IF 2.2) Pub Date : 2024-03-13 Jiaqi Zheng, Yuchuan Zhang, Yiting Cai, Wei Han, Wei Chen
-
Selection of positive controls and their impact on anti-drug antibody assay performance J. Immunol. Methods (IF 2.2) Pub Date : 2024-03-11 Joshua A. Weiner, Harini Natarajan, Calum J. McIntosh, Eun Sung Yang, Misook Choe, Cassidy L. Papia, Katherine S. Axelrod, Gabriela Kovacikova, Amarendra Pegu, Margaret E. Ackerman
Development of assays to reliably identify and characterize anti-drug antibodies (ADAs) depends on positive control anti-idiotype (anti-id) reagents, which are used to demonstrate that the standards recommended by regulatory authorities are met. This work employs a set of therapeutic antibodies under clinical development and their corresponding anti-ids to investigate how different positive control
-
A flow cytometry-based assay to determine the ability of anti-Streptococcus pyogenes antibodies to mediate monocytic phagocytosis in human sera J. Immunol. Methods (IF 2.2) Pub Date : 2024-03-06 Elena Boero, Martina Carducci, Alexander J. Keeley, Francesco Berlanda Scorza, Miren Iturriza-Gómara, Danilo Gomes Moriel, Omar Rossi
-
Development and basic performance verification of a rapid homogeneous bioassay for agonistic antibodies against the thyroid-stimulating hormone receptor J. Immunol. Methods (IF 2.2) Pub Date : 2024-03-05 Motoki Hoshina, Shiomi Ojima, Atsushi Kawasaki, Kosuke Doi, Satoshi Ohta, Asuka Inoue, Hiroshi Murayama
Graves' disease is a type of autoimmune hyperthyroidism caused by thyroid-stimulating antibodies (TSAb). The combination of a porcine thyroid cell bioassay and cyclic adenosine monophosphate (cAMP) immunoassay (TSAb-enzyme immunoassay; EIA) is a clinically approved TSAb measurement method. Due to the requirement of multiple procedures and a long assay time of 6 h in the TSAb-EIA, a simplified and rapid
-
Performance of Softcup® menstrual cup and vulvovaginal swab samples for detection and quantification of genital cytokines J. Immunol. Methods (IF 2.2) Pub Date : 2024-03-05 Nashlin Pillay, Gugulethu Favourate Mzobe, Marothi Letsoalo, Asavela Olona Kama, Andile Mtshali, Stanley Nzuzo Magini, Nikkishia Singh, Vani Govender, Natasha Samsunder, Megeshinee Naidoo, Dhayendre Moodley, Cheryl Baxter, Derseree Archary, Sinaye Ngcapu
Cytokines are important mediators of immunity in the female genital tract, and their levels may be associated with various reproductive health outcomes. However, the measurement of cytokines and chemokines in vaginal fluid samples may be influenced by a variety of factors, each with the potential to affect the sensitivity and accuracy of the assay, including the interpretation and comparison of data
-
An immunochromatographic test for serological diagnosis of scrub typhus J. Immunol. Methods (IF 2.2) Pub Date : 2024-03-01 Shuhao Yan, Qingyu Lu, Qingyuan Tao, Yawei Lu, Bao Gao, Sibo Wang, Xusheng Cai, Lele Ai, Xiaohui Xiong, Min Cao, Weilong Tan
A fluorescent immunochromatographic test (FM-ICT) was developed for rapid detection of anti- antibodies in serum samples. The FM-ICT was constructed based on the dual-antigen sandwich method. Truncated 56 kDa outer membrane protein of strain SJ, was expressed in and mixed with those of Ptan and Gillam strains. A thin line of the protein mixture was precisely sprayed across a nitrocellulose membrane
-
High-resolution epitope mapping of commercial antibodies to ANCA antigens by yeast surface display J. Immunol. Methods (IF 2.2) Pub Date : 2024-03-01 John S. Poulton, Sajan Lamba, Meghan Free, Gang Xi, Elizabeth McInnis, Gabrielle Williams, Stephan T. Kudlacek, David Thieker, Brian Kuhlman, Ronald Falk
Epitope mapping provides critical insight into antibody-antigen interactions. Epitope mapping of autoantibodies from patients with autoimmune diseases can help elucidate disease immunogenesis and guide the development of antigen-specific therapies. Similarly, epitope mapping of commercial antibodies targeting known autoantigens enables the use of those antibodies to test specific hypotheses. Anti-Neutrophil
-
A comprehensive method for the phenotypical and functional characterization of recalled human memory B and T cells specific to vaccine antigens J. Immunol. Methods (IF 2.2) Pub Date : 2024-02-28 Czdari Lee, Imtisal Imran, Sara Thomas, Mahyar Nouri-Shirazi
-
Methodological optimisation of thymocyte isolation and cryopreservation of human thymus samples J. Immunol. Methods (IF 2.2) Pub Date : 2024-02-27 Ruth R. Hagen, Calvin Xu, Hui-Fern Koay, Igor E. Konstantinov, Stuart P. Berzins, Katherine Kedzierska, Carolien E. van de Sandt
Premature lymphocytes develop into non-autoreactive, mature naïve CD4 or CD8 T cells in the thymus before entering the circulation. However, in-depth characterization of human thymocyte development remains challenging due to limited availability of human thymus samples and the fragile nature of thymocyte populations. Thymocytes often do not survive cryopreservation and thawing procedures, especially
-
True volumetric counting of CD34+ cells using flow cytometry J. Immunol. Methods (IF 2.2) Pub Date : 2024-02-21 Laura G. Rico, Jorge Bardina, Roser Salvia, Michael D. Ward, Jolene A. Bradford, Jordi Petriz
While the single-platform flow cytometric CD34+ cell counting method is the preferred choice to predict the yield of mobilized peripheral blood stem cells, most flow cytometers lack the ability of hematology counter analyzers to perform volumetric counting. However, one of the problems using reference microbeads is the vanishing counting bead phenomenon. This phenomenon results in a drop in microbeads
-
-
Development of gold nanocluster complex for the detection of tumor necrosis factor-alpha based on immunoassay J. Immunol. Methods (IF 2.2) Pub Date : 2024-02-18 Natchanok Talapphet, Chang Soon Huh, Moon-Moo Kim
Tumor necrosis factor-alpha, TNF-α, a cytokine recognized as a key regulator of inflammatory responses, is primarily produced by activated monocytes and macrophages. Measuring TNF-α levels serves as a valuable indicator for tracking several diseases and pathological states. Gold nanotechnology has been identified as a highly effective catalyst with unique properties for measuring inflammatory cytokines
-
Comparative assessment of cytometry by time-of-flight and full spectral flow cytometry based on a 33-color antibody panel J. Immunol. Methods (IF 2.2) Pub Date : 2024-02-15 Antonia Schäfer, Sènan Mickael D'Almeida, Julien Dorier, Nicolas Guex, Jean Villard, Miguel Garcia
Mass cytometry and full spectrum flow cytometry have recently emerged as new promising single cell proteomic analysis tools that can be exploited to decipher the extensive diversity of immune cell repertoires and their implication in human diseases. In this study, we evaluated the performance of mass cytometry against full spectrum flow cytometry using an identical 33-color antibody panel on four healthy
-
Comparison of real-time quantitative PCR and two digital PCR platforms to detect copy number variation in FCGR3B J. Immunol. Methods (IF 2.2) Pub Date : 2024-02-06 Kirstine Kløve-Mogensen, Simone Karlsson Terp, Rudi Steffensen
The importance of structural genetic variants, such as copy number variations (CNVs), in modulating human disease is being increasingly recognized. Several clinical conditions require investigation of human neutrophil antigen (HNA-1), which is encoded by the Fc gamma receptor IIIb gene (, including suspicion of neutropenia, infections, and proactive testing of blood component donors to reduce the potential
-
Methods for studying P2X4 receptor ion channels in immune cells J. Immunol. Methods (IF 2.2) Pub Date : 2024-02-03 Ronald Sluyter, Tahnee B.-D. McEwan, Reece A. Sophocleous, Leanne Stokes
The P2X4 receptor is a trimeric ligand-gated ion channel activated by adenosine 5′-triphosphate (ATP). P2X4 is present in immune cells with emerging roles in inflammation and immunity, and related disorders. This review aims to provide an overview of the methods commonly used to study P2X4 in immune cells, focusing on those methods used to assess gene expression, the presence of the P2X4 protein, and
-
Comparison of three methods for the detection of antibodies against muscle-specific kinase J. Immunol. Methods (IF 2.2) Pub Date : 2024-02-03 Kyphuong Luong, Bucky K. Lozier, Camille L. Novis, Tammy L. Smith, Lauren M. Zuromski, Lisa K. Peterson
To compare 3 different methods for the detection of antibodies against muscle-specific kinase (MuSK). MuSK antibody testing was performed in 237 serum samples by enzyme-linked immunosorbent assay (ELISA) and fixed cell-based assay (f-CBA-IFA). One hundred and forty-eight (148) of the sera had previously been tested by RIA during clinical testing: 47 MuSK antibody positive and 101 MuSK antibody negative
-
-
Immunofluorescence analysis of human eosinophils J. Immunol. Methods (IF 2.2) Pub Date : 2024-01-23 Sarah Almas, Nawell Fayad, Ojas Srivastava, Mujtaba Siddique, Sharanya Das, Nicolas Touret, Xuejun Sun, Paige Lacy
A prominent inflammatory cell type in allergic diseases is the eosinophil, a granulated white blood cell that releases pro-inflammatory cytokines. Eosinophil-derived cytokines, including interleukin-9 (IL-9) and interleukin-13 (IL-13), can skew the immune response towards an allergic phenotype. Unfortunately, it is challenging to immunolabel and collect quantifiable images of eosinophils given their
-
Development and characterization of a hemolysis inhibition assay to determine functionality of anti-Streptolysin O antibodies in human sera J. Immunol. Methods (IF 2.2) Pub Date : 2024-01-16 Martina Carducci, Alana Whitcombe, Luca Rovetini, Luisa Massai, Alexander J. Keeley, Thushan I. de Silva, Julie Bennett, Francesco Berlanda Scorza, Miren Iturriza, Nicole J. Moreland, Danilo G. Moriel, Omar Rossi
The high burden of disease and the long-lasting sequelae following Streptococcus pyogenes (Strep A) infections make the development of an effective vaccine a global health priority. Streptolysin O (SLO), is a key toxin in the complex pathogenesis of Strep A infection. Antibodies are elicited against SLO after natural exposure and represent a key target for vaccine-induced immunity. Here we present
-
Miniaturized CAR knocked onto CD3ε extends TCR function with CAR specificity under control of endogenous TCR signaling cascade J. Immunol. Methods (IF 2.2) Pub Date : 2024-01-11 Katrin Manske, Lisa Dreßler, Simon P. Fräßle, Manuel Effenberger, Claudia Tschulik, Vlad Cletiu, Eileen Benke, Michaela Wagner, Kilian Schober, Thomas R. Müller, Christian Stemberger, Lothar Germeroth, Dirk H. Busch, Mateusz P. Poltorak
Immunotherapy using TCR and especially CAR transgenic T cells is a rapidly advancing field with the potential to become standard of care for the treatment of multiple diseases. While all current FDA approved CAR T cell products are generated using lentiviral gene transfer, extensive work is put into CRISPR/Cas mediated gene delivery to develop the next generation of safer and more potent cell products
-
Stability of anti-drug antibodies in human serum samples J. Immunol. Methods (IF 2.2) Pub Date : 2024-01-10 Helene Solberg, Christina Bartholdy, Lone Hummelshøj Landsy, Lars Ole Andresen
Stability of anti-drug antibodies (ADAs) is important as ADA-analysis should be reliable over time at different storage conditions. Stability of anti-insulin antibodies in serum samples was assessed after short-term storage at different temperatures and after long-term storage at −20 °C. Correlation between measurements was tested and acceptance criteria for incurred sample reanalysis were applied
-
Development of optimized cytotoxicity assays for assessing the antitumor potential of CAR-T cells J. Immunol. Methods (IF 2.2) Pub Date : 2023-12-24 Misa Eugene-Norbert, Alexis Cuffel, Gaetan Riou, Laetitia Jean, Clara Blondel, Justine Dehayes, Aurélie Bisson, Camille Giverne, Emilie Brotin, Christophe Denoyelle, Laurent Poulain, Olivier Boyer, Jérémie Martinet, Jean-Baptiste Latouche
CAR-T cells are T cells expressing a chimeric antigen receptor (CAR) rendering them capable of killing tumor cells after recognition of a target antigen. CD19 CAR-T cells have revolutionized the treatment of hematological malignancies. Their function is typically assessed by cytotoxicity assays using human allogeneic cell lines expressing the target antigen CD19 such as Nalm-6. However, an alloreactive
-
Antibody engineering to generate anti-tumor-associated glycoprotein 72 mouse recombinant CC49 IgG with improved solubility, purity, and thermal stability J. Immunol. Methods (IF 2.2) Pub Date : 2023-12-23 Zhihong Lin, Bailin Tu, Philip M. Hemken, A. Scott Muerhoff
Tumor-associated glycoprotein 72 (TAG-72) is a mucin that is overexpressed heterogeneously on the surface of cancer cells, and is a potential target for immunotherapies for various cancer types. As a tumor marker, TAG-72 is measured with the cancer antigen (CA) 72–4 immunoassay. The murine monoclonal antibody (mAb) CC49 is a second-generation IgG that targets an antigen on TAG-72; however, CC49 has
-
A live mammalian cells electroporation array for on-chip immunofluorescence J. Immunol. Methods (IF 2.2) Pub Date : 2023-12-23 Marta Maschietto, Stefano Girardi, Onelia Gagliano, Stefano Vassanelli
The detection of intracellular proteins in vitro is commonly realized with immunofluorescence techniques, through which antibodies or markers are delivered into fixed cells and recognize specific proteins. Many innovative techniques, however, avoid cells fixation by chemical compounds and, among the others, electroporation is widely used. Here we demonstrate that in situ electroporation on thin film
-
Dissociation of murine oral mucosal tissues for single cell applications J. Immunol. Methods (IF 2.2) Pub Date : 2023-12-22 Tomoko Ikeuchi, Ramin Akhi, Belmaliz Cardona Rodriguez, David Fraser, Drake Williams, Tae Sung Kim, Teresa Greenwell-Wild, Andrew Overmiller, Maria Morasso, Niki Moutsopoulos
Single-cell RNA sequencing and flow cytometry approaches have been instrumental in understanding cellular states within various tissues and organs. However, tissue dissociation methods can potentially alter results and create bias due to preferential recovery of particular cell types. Here we present efforts to optimize methods for dissociation of murine oral mucosal tissues and provide three different
-
Novel hapten design, highly sensitive monoclonal antibody production, and immunoassay development for rapid screening of illegally added chloramphenicol in cosmetics J. Immunol. Methods (IF 2.2) Pub Date : 2023-12-22 Zhaoxiang Wang, Mian Wang, Xiaoxiang Fu, Jingqi Qian, Min Wang, Guiyu Tan
Hapten design and synthesis have been regarded as the key factor to generate high-quality antibodies. In the present study, a novel hapten of chloramphenicol was synthesized, characterized and compared with two conventional haptens. The new hapten generated mAb 4B5 showed higher sensitivity and titer than the other two haptens-based mAbs. The haptens synthesized with the structure of chloramphenicol
-
Tumor nano-lysate activates dendritic cells to evoke a preventative immune response J. Immunol. Methods (IF 2.2) Pub Date : 2023-12-12 Jenna A. Dombroski, Abigail R. Fabiano, Samantha V. Knoblauch, Schyler J. Rowland, Katherine N. Gibson-Corley, Michael R. King
A tumor nano-lysate “TNL” vaccine comprised of sonicated 4T1 cells was developed, characterized and implemented for the prevention of triple-negative breast cancer. This study aimed to gain a better understanding of the immune response behind the success of the vaccine in vivo, through use of ex vivo and in vivo assays. Here, we analyze the activation of various immune cells isolated from healthy mouse
-
A SMART method for isolating monoclonal antibodies from individual rhesus macaque memory B cells J. Immunol. Methods (IF 2.2) Pub Date : 2023-12-14 Jason T. Weinfurter, Sarah N. Bennett, Matthew R. Reynolds
-
Variegate expression of Cre recombinase in hematopoietic cells in CD11c-cre transgenic mice J. Immunol. Methods (IF 2.2) Pub Date : 2023-12-14 Claire Murat, Sylvie Guerder
In this study, we performed an in-depth analysis of Cre expression in the widely used CD11c-Cre transgenic mice generated by the group of Boris Reizis. In contrast to previous observation, using the highly sensitive Rosa-26-floxed-tdTomato reporter mouse line, we show variegated expression of Cre in multiple hematopoietic linage cells starting in hematopoietic stem cells. Indeed, we found that in the
-
Application of enzyme-linked immunosorbent assay to detect antimicrobial peptides in human intestinal lumen J. Immunol. Methods (IF 2.2) Pub Date : 2023-12-09 Julie S. Hong, Abrar Shamim, Hussein Atta, Eric B. Nonnecke, Sarah Merl, Satyajit Patwardhan, Elin Manell, Esad Gunes, Philip Jordache, Bryan Chen, Wuyuan Lu, Bo Shen, Beatrice Dionigi, Ravi P. Kiran, Megan Sykes, Emmanuel Zorn, Charles L. Bevins, Joshua Weiner
Intestinal transplantation is the definitive treatment for intestinal failure. However, tissue rejection and graft-versus-host disease are relatively common complications, necessitating aggressive immunosuppression that can itself pose further complications. Tracking intraluminal markers in ileal effluent from standard ileostomies may present a noninvasive and sensitive way to detect developing pathology
-
In silico epitope prediction of Borrelia burgdorferi sensu lato antigens for the detection of specific antibodies J. Immunol. Methods (IF 2.2) Pub Date : 2023-12-07 Weronika Grąźlewska, Karolina Sołowińska, Lucyna Holec-Gąsior
Despite many years of research, serodiagnosis of Lyme disease still faces many obstacles. Difficulties arise mainly due to the low degree of amino acid sequence conservation of the most immunogenic antigens among B. burgdorferi s.l. genospecies, as well as differences in protein production depending on the environment in which the spirochete is located. Mapping B-cell epitopes located on antigens allows
-
Development of automated microfluidic immunoassays for the detection of SARS-CoV-2 antibodies and antigen J. Immunol. Methods (IF 2.2) Pub Date : 2023-11-30 Linwood Johnson, Maggie L. Bartlett, Francisco Ramirez, Christopher D. Heger, Darci R. Smith
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the coronavirus disease 2019 (COVID-19) global pandemic. Rapid and sensitive detection of the virus soon after infection is important for the treatment and prevention of transmission of COVID-19, and detection of antibodies is important for epidemiology, assessment of vaccine immunogenicity, and identification of
-
Development of a high-throughput image cytometric screening method as a research tool for immunophenotypic characterization of patient samples from clinical studies J. Immunol. Methods (IF 2.2) Pub Date : 2023-11-29 Samir Patel, James I. McDonald, Hamza Mohammed, Vaishnavi Parthasarathy, Veronica Hernandez, Tyanna Stuckey, Allen H. Lin, Srinivas Koushik Gundimeda, Bo Lin, Julian Reading, Leo Li-Ying Chan
Immunophenotyping has been the primary assay for characterization of immune cells from patients undergoing therapeutic treatments in clinical research, which is critical for understanding disease progression and treatment efficacy. Currently, flow cytometry has been the dominant methodology for characterizing surface marker expression for immunological research. Flow cytometry has been proven to be
-
Evaluation of HLA-DR expression in monocytes and CD64 in neutrophils as A predictor of SEPSIS/sirs in the infectious-inflammatory process J. Immunol. Methods (IF 2.2) Pub Date : 2023-12-01 Clara Giffoni de Freitas, Mariela Granero Farias
Sepsis is a highly fatal disease that affects millions of people worldwide every year. Currently, the diagnosis of sepsis is made by identifying at least two symptoms of systemic inflammatory response syndrome (SIRS), along with confirming the presence of microorganisms using a blood culture examination. Some biomarkers are already used to aid in the diagnosis, such as increased levels of C-reactive
-
Validation of a double-color ELISpot assay of IFN-γ and IL-4 production in human peripheral blood mononuclear cells J. Immunol. Methods (IF 2.2) Pub Date : 2023-11-29 Francesca Dapporto, Domiziana De Tommaso, Camilla Marrocco, Pietro Piu, Claudia Semplici, Giulia Fantoni, Ilaria Ferrigno, Giulia Piccini, Martina Monti, Francesca Vanni, Ilaria Razzano, Ilaria Manini, Emanuele Montomoli, Alessandro Manenti
The Enzyme-Linked ImmunoSpot (ELISpot) assay detects cytokines secreted during T cell-specific immune responses against pathogens. As this assay has acquired importance in the clinical setting, standard bioanalytical evaluation of this method is required. Here, we describe a formal bioanalytical validation of a double-color ELISpot assay for the evaluation of IFN-γ and IL-4 released by T helper 1 and
-
Development of a proficiency testing program for HPV serology assays used to evaluate antibody responses in vaccine trials J. Immunol. Methods (IF 2.2) Pub Date : 2023-11-09 Cheryl Miller, Troy J. Kemp, Ligia A. Pinto
In this study, a proficiency panel was created for evaluation of assay performance and inter- and intra-laboratory assay comparisons, especially the ability to accurately measure negative, low, intermediate, and high levels of HPV type-specific antibodies. Comprised of 80 deidentified samples, this panel is designed for individual labs to evaluate assay performance characteristics on a biennial basis
-
Proficiency tests to evaluate the impact on assay outcomes of harmonized influenza-specific Intracellular Cytokine Staining (ICS) and IFN-ɣ Enzyme-Linked ImmunoSpot (ELISpot) protocols J. Immunol. Methods (IF 2.2) Pub Date : 2023-10-31 Gwenn Waerlop, Geert Leroux-Roels, Anke Pagnon, Sarah Begue, Bruno Salaun, Michel Janssens, Donata Medaglini, Elena Pettini, Emanuele Montomoli, Elena Gianchecchi, Teresa Lambe, Leila Godfrey, Maireid Bull, Duncan Bellamy, Håkon Amdam, Geir Bredholt, Rebecca Jane Cox, Frédéric Clement
The magnitude and quality of cell-mediated immune responses elicited by natural infection or vaccination are commonly measured by Interferon-ɣ (IFN-ɣ) Enzyme-Linked ImmunoSpot (ELISpot) and Intracellular Cytokine Staining (ICS). To date, laboratories apply a variety of in-house procedures which leads to diverging results, complicates interlaboratory comparisons and hampers vaccine evaluations. During
-
Yclon: Ultrafast clustering of B cell clones from high-throughput immunoglobulin repertoire sequencing data J. Immunol. Methods (IF 2.2) Pub Date : 2023-10-30 João Gervásio, Alice Ferreira, Liza F. Felicori
Motivation The next-generation sequencing technologies have transformed our understanding of immunoglobulin (Ig) profiles in various immune states. Clonotyping, which groups Ig sequences into B cell clones, is crucial in investigating the diversity of repertoires and changes in antigen exposure. Despite its importance, there is no widely accepted method for clonotyping, and existing methods are computationally
-
A flexible gradient lateral flow immunochromatographic assay for qualitative, semi-quantitative, and quantitative determination of serum amyloid A J. Immunol. Methods (IF 2.2) Pub Date : 2023-10-24 Guangtian Yang, Jishun Li, Shenglan Zhang, Huixiang Ouyang, Chunhai Jiang, Hongcheng Pan
Serum amyloid A (SAA) is an acute-phase protein produced in response to inflammatory proteins during infections, inflammation, trauma, surgery, cancer, and other conditions. Early and accurate detection of SAA is necessary for diagnosis and monitoring of disease progression. To meet this need, we developed a gradient lateral flow immunoassay test strip using Au nanoparticles as signal reporters. The
-
Assessment of mimicking by EBV-CMV immunoglobulin M of anti-HLA antibodies J. Immunol. Methods (IF 2.2) Pub Date : 2023-10-20 Rasime Derya Güleç, Fatma Demet Arslan, Berk Özyılmaz, Nisel Yılmaz, Sevgi Yılmaz Hancı, Şükran Köse
Objective We aimed to show the cross-reactivity that may occur between immunoglobulin (Ig) M antibodies that form against Cytomegalovirus (CMV) and/or Epstein-Barr virus (EBV) and human leukocyte antigens (HLA). Methods Complement-dependent cytotoxicity (CDC) cross-reactivity between serum samples of 57 patients with IgM positive CMV and/or EBV infections and T and B cells from 15 healthy donors were
-
A competitive ligand binding assay for detection of neutralizing antibodies against an insulin analog J. Immunol. Methods (IF 2.2) Pub Date : 2023-10-14 Victoria L. Peek, Deven M. Lemen, Robert J. Konrad, Yi Wen
Biotherapeutics have the potential to trigger undesired immune responses in the patients. For therapeutic proteins, immunogenicity is manifested as anti-drug antibodies (ADA). Because ADA could compromise pharmacokinetics, efficacy, and safety, regulatory agencies require immunogenicity assessment during clinical development. A tiered bioanalytical approach is recommended to monitor clinical immunogenicity
-
Evaluation of multiple immunoassay formats for detection of anti-drug antibodies to zinpentraxin alfa J. Immunol. Methods (IF 2.2) Pub Date : 2023-10-09 Xiaoyun Yang, Ketevan Siradze, Gizette Sperinde, Audrey Arjomandi, Saloumeh Fischer
Zinpentraxin alfa (rhPTX-2; PRM-151) is currently being developed for the treatment of fibrotic diseases such as idiopathic pulmonary fibrosis and myelofibrosis. Notably, because it is administered chronically and has an endogenously expressed counterpart, clinical studies of zinpentraxin alpha must include immunogenicity assessments. Since the typical homogenous bridging ELISA assay does not adequately
-
Impact of bridge heterology on functional parameters of ELISA for 17α-methyltestosterone J. Immunol. Methods (IF 2.2) Pub Date : 2023-09-28 Divya Verma, T.G. Shrivastav, Sonu Chand Thakur
-
Diagnosis of septic shock by serum measurement of human neutrophil lipocalin by a rapid homogeneous assay J. Immunol. Methods (IF 2.2) Pub Date : 2023-09-27 Huijun Zong, Xueyi Shang, Xin Wang, Ting Chen, Ye Wang, Yuhao Ren, Yongqiang Jiang, Yan Li, Qingyu Lv, Peng Liu
Background Human neutrophil lipocalin (HNL) is a marker of neutrophil activation and has a high efficacy in diagnosing bacterial infections. In this study, we applied the AlphaLISA technique to measure the serum level of HNL, evaluate HNL's efficacy in diagnosing septic shock, and identify any association between HNL level and septic patients' prognosis. Methods We collected 146 serum samples from
-
A systematic examination of anti-drug antibody titer estimation: Applied recommendations J. Immunol. Methods (IF 2.2) Pub Date : 2023-09-23 Lin Zhang, Michael E. Hodsdon, Thomas Pottanat, Sihe Wang, Robert John Konrad, Nicholas Seta, Richard E. Higgs
Biologic drugs (therapeutic proteins or peptides) have become one of the most important therapeutic modalities over the past few decades. Drug-induced immunogenicity is a significant concern as it may affect safety, tolerability, and efficacy. With more sensitive and drug-tolerant screening assays in use today, reliable estimation of anti-drug-antibody (ADA) titer has become more important for understanding
-
Standardization, validation, and comparative evaluation of a faster and high-performance test for quantification of yellow fever neutralizing antibodies J. Immunol. Methods (IF 2.2) Pub Date : 2023-09-23 Marisol Simões, Stephanie Almeida da Silva, Kelly Araújo Lúcio, Renan de Oliveira Vieira, Waleska Dias Schwarcz, Sheila Maria Barbosa de Lima, Luiz Antonio Bastos Camacho
Although it is considered the reference for quantification of neutralizing antibodies, classical method of the plaque reduction neutralization test (PRNT) is labor intensive, requires specific equipment and inputs, besides a long time for its finalization, even in the micro-PRNT version (in 96-well plates). It has a higher sample throughput, however the smaller wells make the reading of plaques more
-
Validation of the use of dried blood spots in a chikungunya virus IgG serological assay J. Immunol. Methods (IF 2.2) Pub Date : 2023-09-22 Tereza Magalhaes, Moyra M. Portilho, Patricia S.S. Moreira, Milena L. Marinho, Wiler P. Dias, Natália M. Gonçalves, Osiyallê A.S. Rodrigues, Jane Montes, Leila Reis, Dilma F. Jesus, Tarcísio O. Silva, Lua S. Dultra, Joilda S. Nery, Guilherme S. Ribeiro
Dried blood spot (DBS) sampling is a simple, fast, and minimally invasive blood collection method that is particularly useful for diagnostic or epidemiological studies in hard-to-reach populations. Nevertheless, the use of DBS in assays that have been optimized with gold-standard samples (serum or plasma) must be optimized to yield reliable results. Here, we describe the validation of DBS in a commercial
-
Obtaining a high titer of polyclonal antibodies from rats to the SARS-CoV-2 nucleocapsid protein and its N- and C-terminal domains for diagnostic test development J. Immunol. Methods (IF 2.2) Pub Date : 2023-09-11 Michelle Teixeira de Almeida, Ana Paula Barbosa, Camila Gasque Bomfim, Aline Biazola Visnardi, Tania Churasacari Vinces, Alexandre Ceroni, Edison Luiz Durigon, Cristiane Rodrigues Guzzo
-
Kinetic titration method in flow cytometry for quantification of cell receptors J. Immunol. Methods (IF 2.2) Pub Date : 2023-09-02 Irina V. Khalo, Andrei A. Karpenko, Viktoriya S. Kozyreva, Anna N. Shilova, Olga A. Abubakirova, Dmitry I. Strokotov, Vyacheslav M. Nekrasov, Valeri P. Maltsev, Andrei V. Chernyshev
For the quantitative determination of cell receptors by fluorescence flow cytometry, we proposed a new method, which takes into account the reaction kinetics. The binding reaction of the ligand with receptors begins after placing the cells in the ligand solution. In the proposed method, there are several samples with the same concentration of cells and different initial concentrations of fluorescently
-
Recombinant antigen 5 from Polybia paulista wasp venom (Hymenoptera, Vespidae): Antigen-specific antibody production and functional profile of CD4+ T cells in the immune response J. Immunol. Methods (IF 2.2) Pub Date : 2023-09-07 Isabela Oliveira Sandrini Assugeni, Murilo Luiz Bazon, Lucas Machado Pinto, Leonardo Antônio Benedito Mainente, Márcia Regina Brochetto-Braga, Ricardo de Lima Zollner, Luís Gustavo Romani Fernandes
Polybia paulista is a neotropical social wasp related to severe accidents and allergic reactions cases, including anaphylaxis, in southeastern Brazil. Antigen 5 (Poly p 5) is a major allergenic protein from its venom with potential use for component-resolved diagnostic. Therefore, the previous characterization of the immune response profile triggered by Poly p 5 should be evaluated. Recombinant Poly
-
An enzyme-free method for isolating testicular macrophages from rodent models J. Immunol. Methods (IF 2.2) Pub Date : 2023-09-01 Wei Cai, Yalong Yang
Macrophages are the major type of immune cell in the testis of both humans and rodents. Testicular macrophages (TMs) play critical roles in maintaining the testicular microenvironment, such as Leydig cell-dependent hormone production, spermatogenesis, and immune balance. A substantial number of studies have used rodent models to investigate the functions of TMs with various methods and harvest macrophages
-
Optimisation of SARS-CoV-2 peptide stimulation and measurement of cytokine output by intracellular flow cytometry and bio-plex analysis J. Immunol. Methods (IF 2.2) Pub Date : 2023-09-06 Jemma Victoria Taylor, Emma Louise Callery, Anthony Rowbottom
Our study was conducted to optimise a peptide stimulation and an intracellular cytokine staining protocol, alongside Bio-Plex supernatant analysis, for use in patients who had previously contracted SARS-CoV-2 or received vaccination against this virus in a clinical laboratory setting. Peripheral Blood Mononuclear Cell extraction and cryopreservation allowed for cells to be stored long term and enhanced
-
Development of a concise and reliable method for quantifying the antibody loaded onto lipid nanoparticles modified with Herceptin J. Immunol. Methods (IF 2.2) Pub Date : 2023-09-01 Yusuke Yano, Rui Tada, Nobuhito Hamano, Kenshin Haruta, Tomomi Kobayashi, Masahiro Sato, Yamato Kikkawa, Yoko Endo-Takahashi, Motoyoshi Nomizu, Yoichi Negishi
Antibodies are essential components of the immune system with a wide range of molecular targets. They have been recognized as modalities for treating several diseases and more than 130 approved antibody-based therapeutics are available for clinical use. However, limitations remain associated with its efficacy, tissue permeability, and safety, especially in cancer treatment. Nanoparticles, particularly