Synthesis of a stable isotopically labeled universal surrogate peptide for use as an internal standard in LC-MS/MS bioanalysis of human IgG and Fc-fusion protein drug candidates
| Autor: | Claudio Mapelli, Douglas James Riexinger, Kimberly Voronin, Jianing Zeng, Ving G. Lee, Samuel J. Bonacorsi, Sharon Gong, Nishith Sanghvi, Hao Jiang, Alban Allentoff |
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| Rok vydání: | 2014 |
| Předmět: |
chemistry.chemical_classification
Bioanalysis Dipeptide Pseudoproline Chromatography biology Organic Chemistry Peptide Mass spectrometry Biochemistry Combinatorial chemistry Immunoglobulin G Analytical Chemistry chemistry.chemical_compound Residue (chemistry) chemistry Drug Discovery Peptide synthesis biology.protein Radiology Nuclear Medicine and imaging Spectroscopy |
| Zdroj: | Journal of Labelled Compounds and Radiopharmaceuticals. 57:579-583 |
| ISSN: | 0362-4803 |
| Popis: | The synthesis of a 16-residue, stable isotopically labeled peptide is described for use as a LC-MS/MS (Liquid chromatography-mass spectrometry/mass spectrometry) internal standard in bioanalytical studies. This peptide serves as a single universal surrogate peptide capable of quantifying a wide variety of immunoglobulin G and Fc-fusion protein drug candidates in animal species used in pre-clinical drug development studies. An efficient synthesis approach for this peptide was developed using microwave-assisted solid phase peptide synthesis (SPPS) techniques, which included the use of a pseudoproline dipeptide derivative. The corresponding conventional room temperature SPPS was unsuccessful and gave only mixtures of truncated products. Stable-labeled leucine was incorporated as a single residue via manual coupling of commercially available Fmoc-[(13) C6 , (15) N]-l-leucine onto an 11-unit segment followed by automated microwave-assisted elaboration of the final four residues. Using this approach, the desired labeled peptide was prepared in high purity and in sufficient quantities for long-term supplies as a bioanalytical internal standard. The results strongly demonstrate the importance of utilizing both microwave-assisted peptide synthesis and pseudoproline dipeptide techniques to allow the preparation of labeled peptides with highly lipophilic and sterically hindered side-chains. |
| Databáze: | OpenAIRE |
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