Development of Immunocapture-LC/MS Assay for Simultaneous ADA Isotyping and Semiquantitation
Therapeutic proteins and peptides have potential to elicit immune responses resulting in anti-drug antibodies that can pose problems for both patient safety and product efficacy. During drug development immunogenicity is usually examined by risk-based approach along with specific strategies for deve...
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| Format: | Article |
| Language: | English |
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Wiley
2016-01-01
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| Series: | Journal of Immunology Research |
| Online Access: | http://dx.doi.org/10.1155/2016/7682472 |
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| author | Lin-Zhi Chen David Roos Elsy Philip |
| author_facet | Lin-Zhi Chen David Roos Elsy Philip |
| author_sort | Lin-Zhi Chen |
| collection | DOAJ |
| description | Therapeutic proteins and peptides have potential to elicit immune responses resulting in anti-drug antibodies that can pose problems for both patient safety and product efficacy. During drug development immunogenicity is usually examined by risk-based approach along with specific strategies for developing “fit-for-purpose” bioanalytical approaches. Enzyme-linked immunosorbent assays and electrochemiluminescence immunoassays are the most widely used platform for ADA detection due to their high sensitivity and throughput. During the past decade, LC/MS has emerged as a promising technology for quantitation of biotherapeutics and protein biomarkers in biological matrices, mainly owing to its high specificity, selectivity, multiplexing, and wide dynamic range. In fully taking these advantages, we describe here an immunocapture-LC/MS methodology for simultaneous isotyping and semiquantitation of ADA in human plasma. Briefly, ADA and/or drug-ADA complex is captured by biotinylated drug or anti-drug Ab, immobilized on streptavidin magnetic beads, and separated from human plasma by a magnet. ADA is then released from the beads and subjected to trypsin digestion followed by LC/MS detection of specific universal peptides for each ADA isotype. The LC/MS data are analyzed using cut-point and calibration curve. The proof-of-concept of this methodology is demonstrated by detecting preexisting ADA in human plasma. |
| format | Article |
| id | doaj-art-8172abbca2b746ab9f1445945ecc2695 |
| institution | OA Journals |
| issn | 2314-8861 2314-7156 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Immunology Research |
| spelling | doaj-art-8172abbca2b746ab9f1445945ecc26952025-08-20T02:08:27ZengWileyJournal of Immunology Research2314-88612314-71562016-01-01201610.1155/2016/76824727682472Development of Immunocapture-LC/MS Assay for Simultaneous ADA Isotyping and SemiquantitationLin-Zhi Chen0David Roos1Elsy Philip2Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT 06877, USABoehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT 06877, USABoehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT 06877, USATherapeutic proteins and peptides have potential to elicit immune responses resulting in anti-drug antibodies that can pose problems for both patient safety and product efficacy. During drug development immunogenicity is usually examined by risk-based approach along with specific strategies for developing “fit-for-purpose” bioanalytical approaches. Enzyme-linked immunosorbent assays and electrochemiluminescence immunoassays are the most widely used platform for ADA detection due to their high sensitivity and throughput. During the past decade, LC/MS has emerged as a promising technology for quantitation of biotherapeutics and protein biomarkers in biological matrices, mainly owing to its high specificity, selectivity, multiplexing, and wide dynamic range. In fully taking these advantages, we describe here an immunocapture-LC/MS methodology for simultaneous isotyping and semiquantitation of ADA in human plasma. Briefly, ADA and/or drug-ADA complex is captured by biotinylated drug or anti-drug Ab, immobilized on streptavidin magnetic beads, and separated from human plasma by a magnet. ADA is then released from the beads and subjected to trypsin digestion followed by LC/MS detection of specific universal peptides for each ADA isotype. The LC/MS data are analyzed using cut-point and calibration curve. The proof-of-concept of this methodology is demonstrated by detecting preexisting ADA in human plasma.http://dx.doi.org/10.1155/2016/7682472 |
| spellingShingle | Lin-Zhi Chen David Roos Elsy Philip Development of Immunocapture-LC/MS Assay for Simultaneous ADA Isotyping and Semiquantitation Journal of Immunology Research |
| title | Development of Immunocapture-LC/MS Assay for Simultaneous ADA Isotyping and Semiquantitation |
| title_full | Development of Immunocapture-LC/MS Assay for Simultaneous ADA Isotyping and Semiquantitation |
| title_fullStr | Development of Immunocapture-LC/MS Assay for Simultaneous ADA Isotyping and Semiquantitation |
| title_full_unstemmed | Development of Immunocapture-LC/MS Assay for Simultaneous ADA Isotyping and Semiquantitation |
| title_short | Development of Immunocapture-LC/MS Assay for Simultaneous ADA Isotyping and Semiquantitation |
| title_sort | development of immunocapture lc ms assay for simultaneous ada isotyping and semiquantitation |
| url | http://dx.doi.org/10.1155/2016/7682472 |
| work_keys_str_mv | AT linzhichen developmentofimmunocapturelcmsassayforsimultaneousadaisotypingandsemiquantitation AT davidroos developmentofimmunocapturelcmsassayforsimultaneousadaisotypingandsemiquantitation AT elsyphilip developmentofimmunocapturelcmsassayforsimultaneousadaisotypingandsemiquantitation |