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Rapid Immunochromatographic Detection of Serum Anti-α-Galactosidase A Antibodies in Fabry Patients after Enzyme Replacement Therapy.

Nakano S, Tsukimura T, Togawa T, Ohashi T, Kobayashi M, Takayama K, Kobayashi Y, Abiko H, Satou M, Nakahata T, Warnock DG, Sakuraba H, Shibasaki F - PLoS ONE (2015)

Bottom Line: We developed an immunochromatography-based assay for detecting antibodies against recombinant α-galactosidase A proteins in serum.A conventional enzyme-linked immunosorbent assay supported the results.Most of the patients who showed immunopositive reaction exhibited classic Fabry phenotype and harbored gene mutations affecting biosynthesis of α-galactosidase A.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Medical Research, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo156-8506, Japan; Synthera Technologies Co., Ltd., 4-5-1 Honkomagome, Bunkyo-ku, Tokyo 113-0021, Japan.

ABSTRACT
We developed an immunochromatography-based assay for detecting antibodies against recombinant α-galactosidase A proteins in serum. The evaluation of 29 serum samples from Fabry patients, who had received enzyme replacement therapy with agalsidase alpha and/or agalsidase beta, was performed by means of this assay method, and the results clearly revealed that the patients exhibited the same level of antibodies against both agalsidase alpha and agalsidase beta, regardless of the species of recombinant α-galactosidase A used for enzyme replacement therapy. A conventional enzyme-linked immunosorbent assay supported the results. Considering these, enzyme replacement therapy with agalsidase alpha or agalsidase beta would generate antibodies against the common epitopes in both agalsidase alpha and agalsidase beta. Most of the patients who showed immunopositive reaction exhibited classic Fabry phenotype and harbored gene mutations affecting biosynthesis of α-galactosidase A. As immunochromatography is a handy and simple assay system which can be available at bedside, this assay method would be extremely useful for quick evaluation or first screening of serum antibodies against agalsidase alpha or agalsidase beta in Fabry disease with enzyme replacement therapy.

No MeSH data available.


Related in: MedlinePlus

Comparison of anti-GLA antibody levels in clinical samples using ELISA and IC.Sera from 29 Fabry patients (#1–29) and controls (Cont) were assayed by ELISA (left side) and IC (right side) as refer to Table 1. Control exhibits the average value of the data of healthy people (n = 20) used blank ELISA without both Aga-A neither Aga-B. According to the protocol, ELISA was performed using magnetic beads attached with Aga-A or Aga-B as antigens and the anti-GLA antibodies in each 100-fold diluted serum were measured with colorimetric methods of OD 450 nm using AP-labelled-anti-human IgG antibody and the substrate. The same serum were 5-fold diluted and applied to IC for Aga-A (black column) and Aga-B (white column), and judged by the color scale. The all data were shown as mean ± SD.
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pone.0128351.g003: Comparison of anti-GLA antibody levels in clinical samples using ELISA and IC.Sera from 29 Fabry patients (#1–29) and controls (Cont) were assayed by ELISA (left side) and IC (right side) as refer to Table 1. Control exhibits the average value of the data of healthy people (n = 20) used blank ELISA without both Aga-A neither Aga-B. According to the protocol, ELISA was performed using magnetic beads attached with Aga-A or Aga-B as antigens and the anti-GLA antibodies in each 100-fold diluted serum were measured with colorimetric methods of OD 450 nm using AP-labelled-anti-human IgG antibody and the substrate. The same serum were 5-fold diluted and applied to IC for Aga-A (black column) and Aga-B (white column), and judged by the color scale. The all data were shown as mean ± SD.

Mentions: The results of the measurement of anti-GLA antibodies with ELISA and IC were summarized in Table 1, and the result of the comparative analysis for them was shown in Fig 3. The OD values of serum including antibodies against Aga-A (black column) and Aga-B (white column) in ELISA (left panel) were well correlated with their visual scores in IC (right panel). The results of the ELISA using Aga-A were well correlated with those using Aga-B (R2 = 0.9973), as shown in Fig 4A. The relationship between the scores in IC and the OD values in ELISA was also examined. The results demonstrated that the scores for IC using Aga-A were well correlated with the OD values for ELISA using Aga-A (R2 = 0.9799), as shown in Fig 4B. The results of IC using Aga-B were also well correlated with those for ELISA using Aga-B (R2 = 0.931), as shown in Fig 4C.


Rapid Immunochromatographic Detection of Serum Anti-α-Galactosidase A Antibodies in Fabry Patients after Enzyme Replacement Therapy.

Nakano S, Tsukimura T, Togawa T, Ohashi T, Kobayashi M, Takayama K, Kobayashi Y, Abiko H, Satou M, Nakahata T, Warnock DG, Sakuraba H, Shibasaki F - PLoS ONE (2015)

Comparison of anti-GLA antibody levels in clinical samples using ELISA and IC.Sera from 29 Fabry patients (#1–29) and controls (Cont) were assayed by ELISA (left side) and IC (right side) as refer to Table 1. Control exhibits the average value of the data of healthy people (n = 20) used blank ELISA without both Aga-A neither Aga-B. According to the protocol, ELISA was performed using magnetic beads attached with Aga-A or Aga-B as antigens and the anti-GLA antibodies in each 100-fold diluted serum were measured with colorimetric methods of OD 450 nm using AP-labelled-anti-human IgG antibody and the substrate. The same serum were 5-fold diluted and applied to IC for Aga-A (black column) and Aga-B (white column), and judged by the color scale. The all data were shown as mean ± SD.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4470989&req=5

pone.0128351.g003: Comparison of anti-GLA antibody levels in clinical samples using ELISA and IC.Sera from 29 Fabry patients (#1–29) and controls (Cont) were assayed by ELISA (left side) and IC (right side) as refer to Table 1. Control exhibits the average value of the data of healthy people (n = 20) used blank ELISA without both Aga-A neither Aga-B. According to the protocol, ELISA was performed using magnetic beads attached with Aga-A or Aga-B as antigens and the anti-GLA antibodies in each 100-fold diluted serum were measured with colorimetric methods of OD 450 nm using AP-labelled-anti-human IgG antibody and the substrate. The same serum were 5-fold diluted and applied to IC for Aga-A (black column) and Aga-B (white column), and judged by the color scale. The all data were shown as mean ± SD.
Mentions: The results of the measurement of anti-GLA antibodies with ELISA and IC were summarized in Table 1, and the result of the comparative analysis for them was shown in Fig 3. The OD values of serum including antibodies against Aga-A (black column) and Aga-B (white column) in ELISA (left panel) were well correlated with their visual scores in IC (right panel). The results of the ELISA using Aga-A were well correlated with those using Aga-B (R2 = 0.9973), as shown in Fig 4A. The relationship between the scores in IC and the OD values in ELISA was also examined. The results demonstrated that the scores for IC using Aga-A were well correlated with the OD values for ELISA using Aga-A (R2 = 0.9799), as shown in Fig 4B. The results of IC using Aga-B were also well correlated with those for ELISA using Aga-B (R2 = 0.931), as shown in Fig 4C.

Bottom Line: We developed an immunochromatography-based assay for detecting antibodies against recombinant α-galactosidase A proteins in serum.A conventional enzyme-linked immunosorbent assay supported the results.Most of the patients who showed immunopositive reaction exhibited classic Fabry phenotype and harbored gene mutations affecting biosynthesis of α-galactosidase A.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Medical Research, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo156-8506, Japan; Synthera Technologies Co., Ltd., 4-5-1 Honkomagome, Bunkyo-ku, Tokyo 113-0021, Japan.

ABSTRACT
We developed an immunochromatography-based assay for detecting antibodies against recombinant α-galactosidase A proteins in serum. The evaluation of 29 serum samples from Fabry patients, who had received enzyme replacement therapy with agalsidase alpha and/or agalsidase beta, was performed by means of this assay method, and the results clearly revealed that the patients exhibited the same level of antibodies against both agalsidase alpha and agalsidase beta, regardless of the species of recombinant α-galactosidase A used for enzyme replacement therapy. A conventional enzyme-linked immunosorbent assay supported the results. Considering these, enzyme replacement therapy with agalsidase alpha or agalsidase beta would generate antibodies against the common epitopes in both agalsidase alpha and agalsidase beta. Most of the patients who showed immunopositive reaction exhibited classic Fabry phenotype and harbored gene mutations affecting biosynthesis of α-galactosidase A. As immunochromatography is a handy and simple assay system which can be available at bedside, this assay method would be extremely useful for quick evaluation or first screening of serum antibodies against agalsidase alpha or agalsidase beta in Fabry disease with enzyme replacement therapy.

No MeSH data available.


Related in: MedlinePlus