Characteristics of erythropoietin antibodies in patients treated with recombinant human erythropoietin

Our aim was to characterize anti-EPO antibodies in serum samples of the patients treated with erythropoietin. 106 serum samples from the patients treated with erythropoietin (EPO) were collected and assayed. 134 serum samples of patients who did not receive EPO were taken for comparative analysis. T...

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Autores principales:	A. M. Kudryashova, L. N. Nesterenko, G. A. Generalova, T. Yu. Abasheeva, N. A. Mikhailova, O. V. Borisova
Formato:	article
Lenguaje:	RU
Publicado:	SPb RAACI 2020
Materias:	erythropoietin antibodies elisa immunogenicity avidity human serum Immunologic diseases. Allergy RC581-607
Acceso en línea:	https://doaj.org/article/6c3e84e7735946cfb8d648d437560eae
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spelling	oai:doaj.org-article:6c3e84e7735946cfb8d648d437560eae2021-11-18T08:03:49ZCharacteristics of erythropoietin antibodies in patients treated with recombinant human erythropoietin1563-06252313-741X10.15789/1563-0625-COE-1879https://doaj.org/article/6c3e84e7735946cfb8d648d437560eae2020-01-01T00:00:00Zhttps://www.mimmun.ru/mimmun/article/view/1879https://doaj.org/toc/1563-0625https://doaj.org/toc/2313-741XOur aim was to characterize anti-EPO antibodies in serum samples of the patients treated with erythropoietin. 106 serum samples from the patients treated with erythropoietin (EPO) were collected and assayed. 134 serum samples of patients who did not receive EPO were taken for comparative analysis. The anti-EPO antibody detection was performed in ELISA test with rhEPO, by passive capture on ELISA plates, using steptavidin-biotin immunochemical system. Mouse monoclonal antibodies to human IgG, IgG1, IgG2, IgG3 and IgG4 conjugated to horseradish peroxidase were used to detect anti-EPO antibodies, and protein-A peroxidase conjugate was used for quantitative assays. Rabbit anti-human EPO polyclonal antibodies at known concentrations were used as a calibration standard. Six calibration samples at the concentration range of 16-1000 ng/ml were used to plot calibration curves. The lower detection limit was 12 ng/mL, and the quantitative detection limit was 31 ng/ml. Immunochemical capturing led to increasing of total IgG antibody detection by 3.2 times, IgG1 – by 1.1 times IgG2 – by 1.25 times, IgG3 – by 1.5 times, IgG4 – by 1.7 times. Antibodies of mixed isotype were found in most patients. IgG1 or IgG4 antibodies to EPO were determined only in 3 samples. Specific IgM was not detectable among 106 sera samples, whereas total IgG antibodies were detected in 36.8 % of cases. In 34% of sera, their presence was confirmed by detection of at least one of the subclasses. IgG1 antibody was detected in 83.3%; IgG4, in 80.6% of the samples positive for total IgG antibodies. In all cases, IgG2 and/or IgG3 were detected in presence of IgG1 or IgG4 antibodies. The antibody concentration was 3.2 to 35.5 µg/mL in sera from 28 patients, in 8 cases the level of antibodies was > 50 µg/ml, however, being below the limit of quantitative detection in 3 patients. Only 6 samples contained antibodies with avidity index of > 50%. Immunochemical capturing of the antigen led to increased sensitivity for detecting all subclasses of specific antibodies. The specific IgG antibodies to EPO were found in more than 1/3 of serum samples from the patients treated with erythropoietin. Low-avidity antibodies of IgG1 and IgG4 subclasses were determined in most cases.A. M. KudryashovaL. N. NesterenkoG. A. GeneralovaT. Yu. AbasheevaN. A. MikhailovaO. V. BorisovaSPb RAACIarticleerythropoietinantibodieselisaimmunogenicityavidityhuman serumImmunologic diseases. AllergyRC581-607RUMedicinskaâ Immunologiâ, Vol 22, Iss 1, Pp 143-152 (2020)
institution	DOAJ
collection	DOAJ
language	RU
topic	erythropoietin antibodies elisa immunogenicity avidity human serum Immunologic diseases. Allergy RC581-607
spellingShingle	erythropoietin antibodies elisa immunogenicity avidity human serum Immunologic diseases. Allergy RC581-607 A. M. Kudryashova L. N. Nesterenko G. A. Generalova T. Yu. Abasheeva N. A. Mikhailova O. V. Borisova Characteristics of erythropoietin antibodies in patients treated with recombinant human erythropoietin
description	Our aim was to characterize anti-EPO antibodies in serum samples of the patients treated with erythropoietin. 106 serum samples from the patients treated with erythropoietin (EPO) were collected and assayed. 134 serum samples of patients who did not receive EPO were taken for comparative analysis. The anti-EPO antibody detection was performed in ELISA test with rhEPO, by passive capture on ELISA plates, using steptavidin-biotin immunochemical system. Mouse monoclonal antibodies to human IgG, IgG1, IgG2, IgG3 and IgG4 conjugated to horseradish peroxidase were used to detect anti-EPO antibodies, and protein-A peroxidase conjugate was used for quantitative assays. Rabbit anti-human EPO polyclonal antibodies at known concentrations were used as a calibration standard. Six calibration samples at the concentration range of 16-1000 ng/ml were used to plot calibration curves. The lower detection limit was 12 ng/mL, and the quantitative detection limit was 31 ng/ml. Immunochemical capturing led to increasing of total IgG antibody detection by 3.2 times, IgG1 – by 1.1 times IgG2 – by 1.25 times, IgG3 – by 1.5 times, IgG4 – by 1.7 times. Antibodies of mixed isotype were found in most patients. IgG1 or IgG4 antibodies to EPO were determined only in 3 samples. Specific IgM was not detectable among 106 sera samples, whereas total IgG antibodies were detected in 36.8 % of cases. In 34% of sera, their presence was confirmed by detection of at least one of the subclasses. IgG1 antibody was detected in 83.3%; IgG4, in 80.6% of the samples positive for total IgG antibodies. In all cases, IgG2 and/or IgG3 were detected in presence of IgG1 or IgG4 antibodies. The antibody concentration was 3.2 to 35.5 µg/mL in sera from 28 patients, in 8 cases the level of antibodies was > 50 µg/ml, however, being below the limit of quantitative detection in 3 patients. Only 6 samples contained antibodies with avidity index of > 50%. Immunochemical capturing of the antigen led to increased sensitivity for detecting all subclasses of specific antibodies. The specific IgG antibodies to EPO were found in more than 1/3 of serum samples from the patients treated with erythropoietin. Low-avidity antibodies of IgG1 and IgG4 subclasses were determined in most cases.
format	article
author	A. M. Kudryashova L. N. Nesterenko G. A. Generalova T. Yu. Abasheeva N. A. Mikhailova O. V. Borisova
author_facet	A. M. Kudryashova L. N. Nesterenko G. A. Generalova T. Yu. Abasheeva N. A. Mikhailova O. V. Borisova
author_sort	A. M. Kudryashova
title	Characteristics of erythropoietin antibodies in patients treated with recombinant human erythropoietin
title_short	Characteristics of erythropoietin antibodies in patients treated with recombinant human erythropoietin
title_full	Characteristics of erythropoietin antibodies in patients treated with recombinant human erythropoietin
title_fullStr	Characteristics of erythropoietin antibodies in patients treated with recombinant human erythropoietin
title_full_unstemmed	Characteristics of erythropoietin antibodies in patients treated with recombinant human erythropoietin
title_sort	characteristics of erythropoietin antibodies in patients treated with recombinant human erythropoietin
publisher	SPb RAACI
publishDate	2020
url	https://doaj.org/article/6c3e84e7735946cfb8d648d437560eae
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Characteristics of erythropoietin antibodies in patients treated with recombinant human erythropoietin

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