Quantitative immunoassay for mink immunoglobulin in serum and milk
Abstract Background The significance of maternal immunoglobulin G (IgG) for the resistance against a number of infections affecting the health of young mink offspring is not known. Here, we present a validated immunoassay for quantification of mink IgG in serum and milk, using a commercially availab...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-06-01
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| Series: | Acta Veterinaria Scandinavica |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13028-018-0391-7 |
| _version_ | 1818025260879970304 |
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| author | Ronja Mathiesen Mariann Chriél Tina Struve Peter Mikael Helweg Heegaard |
| author_facet | Ronja Mathiesen Mariann Chriél Tina Struve Peter Mikael Helweg Heegaard |
| author_sort | Ronja Mathiesen |
| collection | DOAJ |
| description | Abstract Background The significance of maternal immunoglobulin G (IgG) for the resistance against a number of infections affecting the health of young mink offspring is not known. Here, we present a validated immunoassay for quantification of mink IgG in serum and milk, using a commercially available polyclonal goat anti-ferret IgG antibody cross-reactive with mink IgG as both the catching and the detection antibody, in a sandwich format enzyme linked immunosorbent assay (ELISA). Using this ELISA, serum IgG concentrations was analyzed over time in both mothers and kits in order to establish a correlation between maternal IgG serum concentrations and those of the offspring. Results Intra-assay coefficient of variation (CV) for a serum sample ranged from 2.15 to 5.97% depending on the dilution, while the inter-assay CV ranged from 5.17 to 17.78%. In addition, the range of milk intra-assay CV was 2.71–5.92%, while the range of the inter-assay CV was 4.20–16.03%. Calibrating the ELISA with purified mink IgG (an in-house preparation purified from mink serum) the lower limit of detection was found to be 5 ng/mL for serum and 1 ng/mL for milk. Both serum and milk showed high precision and good linearity over a two-log dilution range. When comparing the serum IgG concentrations of the mink kits a clear within litter effect was seen, while the mean serum IgG concentrations of litters differed significantly between some of the litters (P = 0.0013). Mean maternal serum IgG concentrations correlated positively with the IgG serum concentration of the corresponding offspring sampled over a 3 week period (R2 = 0.63). Conclusions A calibrated and reproducible sandwich ELISA for quantifying mink IgG concentrations in both milk and serum with high analytical sensitivity was developed and validated. The results in this study corroborate previous investigations supporting the usability of the ELISA, paving the way for investigations into the importance of maternal IgG in milk and in serum for the welfare and health of the offspring. |
| first_indexed | 2024-12-10T04:13:17Z |
| format | Article |
| id | doaj.art-7a64f01c279a46d69765840e4ddbda36 |
| institution | Directory Open Access Journal |
| issn | 1751-0147 |
| language | English |
| last_indexed | 2024-12-10T04:13:17Z |
| publishDate | 2018-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Acta Veterinaria Scandinavica |
| spelling | doaj.art-7a64f01c279a46d69765840e4ddbda362022-12-22T02:02:40ZengBMCActa Veterinaria Scandinavica1751-01472018-06-016011910.1186/s13028-018-0391-7Quantitative immunoassay for mink immunoglobulin in serum and milkRonja Mathiesen0Mariann Chriél1Tina Struve2Peter Mikael Helweg Heegaard3Innate Immunology Group, National Veterinary Institute, Technical University of DenmarkDivision of Diagnostics & Scientific Advice-Diagnostic & Development, National Veterinary Institute, Technical University of DenmarkKopenhagen FurInnate Immunology Group, National Veterinary Institute, Technical University of DenmarkAbstract Background The significance of maternal immunoglobulin G (IgG) for the resistance against a number of infections affecting the health of young mink offspring is not known. Here, we present a validated immunoassay for quantification of mink IgG in serum and milk, using a commercially available polyclonal goat anti-ferret IgG antibody cross-reactive with mink IgG as both the catching and the detection antibody, in a sandwich format enzyme linked immunosorbent assay (ELISA). Using this ELISA, serum IgG concentrations was analyzed over time in both mothers and kits in order to establish a correlation between maternal IgG serum concentrations and those of the offspring. Results Intra-assay coefficient of variation (CV) for a serum sample ranged from 2.15 to 5.97% depending on the dilution, while the inter-assay CV ranged from 5.17 to 17.78%. In addition, the range of milk intra-assay CV was 2.71–5.92%, while the range of the inter-assay CV was 4.20–16.03%. Calibrating the ELISA with purified mink IgG (an in-house preparation purified from mink serum) the lower limit of detection was found to be 5 ng/mL for serum and 1 ng/mL for milk. Both serum and milk showed high precision and good linearity over a two-log dilution range. When comparing the serum IgG concentrations of the mink kits a clear within litter effect was seen, while the mean serum IgG concentrations of litters differed significantly between some of the litters (P = 0.0013). Mean maternal serum IgG concentrations correlated positively with the IgG serum concentration of the corresponding offspring sampled over a 3 week period (R2 = 0.63). Conclusions A calibrated and reproducible sandwich ELISA for quantifying mink IgG concentrations in both milk and serum with high analytical sensitivity was developed and validated. The results in this study corroborate previous investigations supporting the usability of the ELISA, paving the way for investigations into the importance of maternal IgG in milk and in serum for the welfare and health of the offspring.http://link.springer.com/article/10.1186/s13028-018-0391-7Mink (Neovison vison)Milk IgGSandwich ELISASerum IgGValidation |
| spellingShingle | Ronja Mathiesen Mariann Chriél Tina Struve Peter Mikael Helweg Heegaard Quantitative immunoassay for mink immunoglobulin in serum and milk Acta Veterinaria Scandinavica Mink (Neovison vison) Milk IgG Sandwich ELISA Serum IgG Validation |
| title | Quantitative immunoassay for mink immunoglobulin in serum and milk |
| title_full | Quantitative immunoassay for mink immunoglobulin in serum and milk |
| title_fullStr | Quantitative immunoassay for mink immunoglobulin in serum and milk |
| title_full_unstemmed | Quantitative immunoassay for mink immunoglobulin in serum and milk |
| title_short | Quantitative immunoassay for mink immunoglobulin in serum and milk |
| title_sort | quantitative immunoassay for mink immunoglobulin in serum and milk |
| topic | Mink (Neovison vison) Milk IgG Sandwich ELISA Serum IgG Validation |
| url | http://link.springer.com/article/10.1186/s13028-018-0391-7 |
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