Development and validation of a cell-based bioassay for the detection of neutralizing antibodies against recombinant human erythropoietin in clinical studies

2004 ◽  
Vol 293 (1-2) ◽  
pp. 115-126 ◽  
Author(s):  
Xin Wei ◽  
Steven J. Swanson ◽  
Shalini Gupta
Keyword(s):  
Clinical Studies ◽  
Recombinant Human Erythropoietin ◽  
Neutralizing Antibodies ◽  
Human Erythropoietin ◽  
A Cell ◽  
Development And Validation

scholarly journals Detection of SARS-CoV-2 neutralizing antibodies with a cell-free PCR assay

2020 ◽  
Author(s):  
Kenneth Danh ◽  
Donna Grace Karp ◽  
Peter V Robinson ◽  
David Seftel ◽  
Mars Stone ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Severe Disease ◽  
Pcr Assay ◽  
Disease Symptoms ◽  
Dna Conjugates ◽  
A Cell ◽  
Pathogen Reduction ◽  
Development And Validation ◽  
The Impact ◽  
Level 2

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to more than 4 million confirmed infections worldwide and over 300,000 deaths. While Remdesivir has recently received FDA emergency use authorization for treatment of SARS-CoV-2 infection, convalescent plasma (CP) with high titers of SARS-CoV-2 neutralizing antibodies (NAbs) from recovered donors remains a promising and widely accessible method to mitigate severe disease symptoms. Here, we describe the development and validation of a cell-free neutralization PCR assay using SARS-CoV-2 spike protein S1 and human ACE2 receptor-DNA conjugates. By comparing with samples collected prior to the outbreak, we confirmed that NAbs were specifically detected in COVID-19 cases. Using our unique assay, the NAb signals are detectable as early as 10 days after onset of symptoms and continue to rise, plateauing after 18 days. Notably, we showed that the use of licensed pathogen reduction technology to inactivate potentially contaminating infectious pathogens in CP did not alter NAb signals, paving a path to safely administer effective CP therapies. The described neutralization PCR assay can serve as a qualification tool to easily identify suitable CP donors of a potentially lifesaving therapy. In addition, this assay tool is readily deployable in standard laboratories with biosafety level 2 capability, and can yield results within 2-3 hr. This advancement can facilitate research on factors driving diverse COVID-19 disease manifestations, and to evaluate the impact of various CP processing protocols on CP therapeutic efficacy.

Development and validation of a cell based assay for the detection of neutralizing antibodies against recombinant insulins

2018 ◽  
Vol 452 ◽  
pp. 53-62 ◽  
Author(s):  
Sanjukta Chatterjee ◽  
Laxmikant Vashishta ◽  
Vinit S. Waichale ◽  
Vivek G. Nayak ◽  
Ramakrishnan Melarkode ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
A Cell ◽  
Development And Validation

scholarly journals Intravenous Epoetin Alfa-epbx versus Epoetin Alfa for Treatment of Anemia in End-Stage Kidney Disease

2018 ◽  
Vol 13 (8) ◽  
pp. 1204-1214 ◽  
Author(s):  
Steven Fishbane ◽  
Bhupinder Singh ◽  
Seema Kumbhat ◽  
Wayne A. Wisemandle ◽  
Nancy E. Martin
Keyword(s):  
Body Weight ◽  
Adverse Events ◽  
Confidence Interval ◽  
Least Squares ◽  
Recombinant Human Erythropoietin ◽  
Neutralizing Antibodies ◽  
Mean Difference ◽  
Epoetin Alfa ◽  
Human Erythropoietin ◽  
Equivalence Margin

Background and objectivesThis study was conducted to compare the safety and efficacy of intravenous epoetin alfa-epbx, an epoetin alfa biosimilar, to epoetin alfa in patients on hemodialysis with ESKD and anemia.Design, setting, participants, & measurementsIn this 24-week, multicenter, double-blind comparative efficacy and safety study, 612 patients on hemodialysis with ESKD and anemia who had stable hemoglobin and were receiving stable doses of intravenous epoetin alfa were randomized (1:1) to intravenous epoetin alfa or epoetin alfa-epbx. Dosing was adjusted according to the epoetin alfa prescribing information. The coprimary efficacy end points were the least squares mean difference between the two treatments in mean weekly hemoglobin level and mean weekly epoetin dose per kilogram of body weight during the last 4 weeks of treatment.ResultsThe least squares mean difference between epoetin alfa-epbx and epoetin alfa in weekly hemoglobin was −0.12 g/dl and the 95% confidence interval (−0.25 to 0.01) was contained within the prespecified equivalence margin (−0.5 to 0.5 g/dl). The least squares mean difference between epoetin alfa-epbx and epoetin alfa in weekly epoetin dose per kilogram of body weight was 0.37 U/kg per week, and the 95% confidence interval (−10.40 to 11.13) was contained within the prespecified equivalence margin (−45 to 45 U/kg per week). Incidences of adverse events (77.1% versus 75.3%), serious adverse events (24.9% versus 27.0%), and deaths (n=5 versus 6) were similar between the epoetin alfa-epbx and epoetin alfa groups, respectively. Five patients tested positive for anti-recombinant human erythropoietin antibodies at baseline, and two additional patients (n=1 per group) developed anti-recombinant human erythropoietin antibodies while on study treatment. All patients tested negative for neutralizing antibodies, and no patient in either group experienced an event of pure red cell aplasia.ConclusionsThis 24-week, comparative, clinical trial in patients on hemodialysis with ESKD and anemia demonstrated there is no clinically meaningful difference in efficacy or safety between epoetin alfa-epbx and epoetin alfa.

scholarly journals Biosimilar recombinant human erythropoietin induces the production of neutralizing antibodies

2011 ◽  
Vol 80 (1) ◽  
pp. 88-92 ◽  
Author(s):  
Kearkiat Praditpornsilpa ◽  
Khajohn Tiranathanagul ◽  
Pawinee Kupatawintu ◽  
Saengsuree Jootar ◽  
Tanin Intragumtornchai ◽  
...  
Keyword(s):  
Recombinant Human Erythropoietin ◽  
Neutralizing Antibodies ◽  
Human Erythropoietin

scholarly journals Modified recombinant human erythropoietin with potentially reduced immunogenicity

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Thanutsorn Susantad ◽  
Mayuree Fuangthong ◽  
Kannan Tharakaraman ◽  
Phanthakarn Tit-oon ◽  
Mathuros Ruchirawat ◽  
...  
Keyword(s):  
Recombinant Human Erythropoietin ◽  
Neutralizing Antibodies ◽  
Structural Integrity ◽  
Computational Design ◽  
Pure Red Cell Aplasia ◽  
Red Cell ◽  
Thai Population ◽  
Mhc Molecules ◽  
Human Erythropoietin

AbstractRecombinant human erythropoietin (rHuEPO) is a biopharmaceutical drug given to patients who have a low hemoglobin related to chronic kidney disease, cancer or anemia. However, some patients repeatedly receiving rHuEPO develop anti-rHuEPO neutralizing antibodies leading to the development of pure red cell aplasia (PRCA). The immunogenic antibody response activated by rHuEPO is believed to be triggered by T-cells recognizing EPO epitopes bound to MHC molecules displayed on the cell surface of APCs. Previous studies have reported an association between the development of anti-rHuEpo-associated PRCA and the HLA-DRB1*09 gene, which is reported to be entrenched in the Thai population. In this study, we used computational design to screen for immunogenic hotspots recognized by HLA-DRB1*09, and predicted seventeen mutants having anywhere between one through four mutations that reduce affinity for the allele, without disrupting the structural integrity and bioactivity. Five out of seventeen mutants were less immunogenic in vitro while retaining similar or slightly reduced bioactivity than rHuEPO. These engineered proteins could be the potential candidates to treat patients who are rHuEpo-dependent and express the HLA-DRB1*09 allele.

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