Validation of the BIACORE 3000 platform for detection of antibodies against erythropoietic agents in human serum samples

Mason, Stephanie; La, Stephon; Mytych, Daniel T.; Swanson, Steven J.; Ferbas, John

doi:10.1185/030079903125002414

Cited by 51 publications

(22 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Where indicated in the figures, antibody concentrations were extrapolated relative to a standard curve created by serial dilution of the human positive control antibody. The concentrations of this antibody were expressed relative to rabbit polyclonal affinitypurified antisera (Amgen, Inc.) (14), since a human anti-epoetin alfa antibody calibrator is not available.…”

Section: Methodsmentioning

confidence: 99%

Feasibility of a Multiplex Flow Cytometric Bead Immunoassay for Detection of Anti-Epoetin Alfa Antibodies

Ferbas

Thomas

Hodgson

et al. 2007

Clin Vaccine Immunol

Self Cite

View full text Add to dashboard Cite

Immunogenicity profiles of recombinant therapeutic proteins are important to understand because antibodies raised against these molecules may have important clinical sequelae. The purpose of the present study was to demonstrate that a flow cytometric bead array could be used to detect clinically relevant antibodies with specificity to such therapeutics. We chose to evaluate well-characterized specimens from persons treated with epoetin alfa that developed antibody-mediated pure red blood cell aplasia as a means to demonstrate the utility of this platform. Our data show that this assay is capable of detecting anti-epoetin alfa antibodies with a relative antibody concentration of 50 ng/ml, where 25 of 25 sera spiked with antibodies at this concentration scored positive. Moreover, the assay was designed to include positive and negative control beads for each specimen that is processed to ensure the specificity of the signal when detected. Measurement of interassay precision supports quantitative estimates of relative antibody concentrations in the range of 313 to 5,000 ng/ml, where the percent coefficient of variation did not exceed 20%. With respect to clinical specimens, antibodies with specificity for epoetin alfa could be easily detected in a set of specimens from persons with pure red blood cell aplasia that had prior exposure to the EPREX brand of recombinant epoetin alfa. Further development and validation of this approach may facilitate successful widespread application of the method for detection of anti-epoetin alfa antibodies, as well as antibodies directed against other recombinant therapeutic proteins.Immune responses that occur in vivo during administration of a therapeutic protein can induce clinically significant adverse events, including neutralization of both the therapeutic and its endogenous counterpart. Although all antibody isotypes may have this property, manifestations appear to emerge after the class switch to the immunoglobulin G (IgG) isotype (11,17). Clinical management of persons with antibody-neutralized endogenous proteins is not straightforward and most commonly involves the cessation of (drug) administration with supportive care on a case-by-case basis until the circulating concentration of antibodies decreases to nonpathological levels (1). The prolonged half-life of antibodies and plasma cells makes this a long-term issue for the patient.A prototypical example of the ability of a recombinant therapeutic protein to raise cross-reactive, neutralizing antibodies is provided by patients that developed antibody-mediated pure red blood cell aplasia during treatment with recombinant epoetin alfa manufactured and sold outside of the United States. This protein was linked to the induction of neutralizing IgG antibodies and the development of pure red cell aplasia (PRCA) in more than 200 patients since 1998 (1), with the majority of cases attributed to the EPREX brand of recombinant epoetin alfa. Even though the manufacturer of EPREX has since taken action to potentially remediate this situ...

show abstract

Section: Methodsmentioning

confidence: 99%

Feasibility of a Multiplex Flow Cytometric Bead Immunoassay for Detection of Anti-Epoetin Alfa Antibodies

Ferbas

Thomas

Hodgson

et al. 2007

Clin Vaccine Immunol

Self Cite

View full text Add to dashboard Cite

show abstract

“…Blood samples taken before the first dose of darbepoetin alfa and at the end of the study were tested for the presence of antibodies to erythropoietic proteins. All antierythropoietic protein antibody assays were done at Amgen using the Biacore 3000 assay [26]. …”

Section: Methodsmentioning

confidence: 99%

Darbepoetin Alfa Effectively Treats Anemia in Patients with Chronic Kidney Disease with de novo Every-Other-Week Administration

et al. 2004

View full text Add to dashboard Cite

Aim: This multicenter, open-label study determined safety and efficacy of once-every-other-week administration of darbepoetin alfa for anemia of chronic kidney disease in erythropoietin-naive patients not on dialysis. Methods: Participants with hemoglobin levels <11.0 g/dl at baseline were administered darbepoetin alfa at an initial dosage of 0.75 µg/kg once every other week. The dose was titrated to achieve and maintain a hemoglobin response, defined as a hemoglobin range of between 11.0 and 13.0 g/dl for up to 24 weeks. The primary end point was the dose of darbepoetin alfa at initial hemoglobin response. Results: Six hundred and eight patients were enrolled, and 463 completed the study; 95% (95% confidence interval: 0.93, 0.97) of the patients who completed treatment achieved a hemoglobin response. The mean darbepoetin alfa dose at the time of response was 63.5 ± (SD) 16.9 µg, and the mean time to hemoglobin response was 5.7 ± (SD) 4.5 weeks. Oral iron therapy was administered to 60% and intravenous iron to 16% of the participants. Darbepoetin alfa was well tolerated, and adverse events were consistent with those expected in patients with chronic kidney disease. Conclusion: Darbepoetin alfa administered once every other week is effective and safe for achieving and maintaining target hemoglobin levels in anemic patients with chronic kidney disease.

show abstract

“…Thus, there is a theoretical possibility to obtain false negative results at a very early stage of the disease. The biosensor immunoassay (BIAcore assay) is a sophisticated assay platform (24) that has recently been applied to the detection of EPO antibodies (25). Briefly, epoetin is immobilized on the surface of a sensorchip, and this biosensor is incubated with serum samples.…”

Section: Assays For Anti-epo Antibodiesmentioning

confidence: 99%

Anti-Erythropoietin Antibodies and Pure Red Cell Aplasia

Rossert¹,

Casadevall²,

Eckardt³

2004

Journal of the American Society of Nephrology

128

View full text Add to dashboard Cite

Validation of the BIACORE 3000 platform for detection of antibodies against erythropoietic agents in human serum samples

Cited by 51 publications

References 11 publications

Feasibility of a Multiplex Flow Cytometric Bead Immunoassay for Detection of Anti-Epoetin Alfa Antibodies

Feasibility of a Multiplex Flow Cytometric Bead Immunoassay for Detection of Anti-Epoetin Alfa Antibodies

Darbepoetin Alfa Effectively Treats Anemia in Patients with Chronic Kidney Disease with de novo Every-Other-Week Administration

Anti-Erythropoietin Antibodies and Pure Red Cell Aplasia

Contact Info

Product

Resources

About