Analytical Investigation of the Profile of Human Chorionic Gonadotropin in Highly Purified Human Menopausal Gonadotrophin Preparations
Abstract
1. Introduction
2. Results
2.1. Intact Molecule Analysis by LC
2.2. Sample Composition Assessed by Intact Molecule Analysis by LC-MS
2.3. Protein Impurity Analysis by LC-MS/MS
2.4. Reducing Peptide Mapping by LC-MS/MS
2.5. Glycopeptide Mapping by LC-MS/MS
2.5.1. Alpha Subunit
2.5.2. Beta Subunits
3. Discussion
4. Materials and Methods
4.1. Samples Used
- Three batches of Menopur® (hFSH and hLH; Ferring Pharmaceuticals, Saint-Prex, Switzerland) (batch number: U12461H, S15094CA, and SI5964CA) HP—freeze-dried.
- Two batches of Menogon® (hFSH and hLH; Ferring Pharmaceuticals, Saint-Prex, Switzerland) (batch number: NU0022A and NU0032A) HP—PEN. Menogon is the same formulation as Menopur® but is sold under a different brand name.
- One batch of Brevactid® (hCG, Ferring Pharmaceuticals, Saint-Prex, Switzerland) (batch number: T14969E). Freeze-dried and contains urinary hCG extracted from the urine of pregnant women.
- One batch of Ovidrel® (r-hCG, Merck KGaA, Darmstadt, Germany) (Batch BCEA17060).
- One batch of pituitary LH standard (Merck KGaA, Darmstadt, Germany) (Batch 869003).
4.2. Glycopeptide Mapping by Hydrophilic Interaction Chromatography (HILIC)–Liquid Chromatography–Mass Spectrometry (LC-MS/MS)
4.3. Intact Molecule Analysis by Reversed-Phase (RP) Liquid Chromatography–Mass Spectrometry (LC-MS)
4.4. Analysis of Impurities by Reversed-Phase (RP) Liquid Chromatography–Mass Spectrometry LC-MS/MS
4.5. Reducing Peptide Mapping by Reversed-Phase (RP) Liquid Chromatography–Mass Spectrometry (LC-MS/MS)
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Species | Control Sample with Mixed Recombinant Gonadotropins | HP-hMG Test Samples | |||
---|---|---|---|---|---|
U12461H | SI5964CA | NU0022A | NU0032A | ||
Beta hCG | 26.04 | 11.44 | 12.26 | 10.27 | 11.37 |
Beta LH | 20.63 | 10.57 | 6.69 | 4.96 | 5.95 |
Impurities | N.D. | 27.71 | 22.76 | 32.23 | 31.53 |
Oxidized species (alpha subunit) | 0.09 | 20.02 | 18.98 | 20.89 | 17.32 |
Samples | Recombinant hCG (Ovidrel®) Batch BCEA17060 | HP-hMG Test Samples | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
U12461H | SI5964CA | NU0022A | NU0032A | |||||||
Replicates | Rep. 1 | Rep. 2 | Rep. 1 | Rep. 2 | Rep. 1 | Rep. 2 | Rep. 1 | Rep. 2 | Rep. 1 | Rep. 2 |
Number of identified proteins | 2 | 5 | 198 | 199 | 200 | 202 | 102 | 91 | 99 | 99 |
Species | Recombinant hCG (Ovidrel®) Batch BCEA17060 | HP-hMG Test Sample | |||
---|---|---|---|---|---|
U12461H | SI5964CA | NU0022A | NU0032A | ||
Alpha | 26.28 | 43.00 | 50.27 | 32.65 | 50.33 |
Beta hCG | 73.73 | 22.33 | 16.93 | 47.38 | 18.03 |
Beta FSH | N.A. | 33.59 | 31.57 | 19.05 | 30.46 |
Beta LH | N.A. | 1.09 | 1.25 | 0.92 | 1.19 |
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Capolupo, A.; Petrocchi, S.; Melchiorre, M.; Jonas, K.; D’Hooghe, T.; Hanyaloglu, A.; Sunkara, S.; Palmese, A.; Ozgumus, B.; Amoresano, A.; et al. Analytical Investigation of the Profile of Human Chorionic Gonadotropin in Highly Purified Human Menopausal Gonadotrophin Preparations. Int. J. Mol. Sci. 2024, 25, 9405. https://doi.org/10.3390/ijms25179405
Capolupo A, Petrocchi S, Melchiorre M, Jonas K, D’Hooghe T, Hanyaloglu A, Sunkara S, Palmese A, Ozgumus B, Amoresano A, et al. Analytical Investigation of the Profile of Human Chorionic Gonadotropin in Highly Purified Human Menopausal Gonadotrophin Preparations. International Journal of Molecular Sciences. 2024; 25(17):9405. https://doi.org/10.3390/ijms25179405
Chicago/Turabian StyleCapolupo, Angela, Sofia Petrocchi, Maura Melchiorre, Kim Jonas, Thomas D’Hooghe, Aylin Hanyaloglu, Sesh Sunkara, Angelo Palmese, Beste Ozgumus, Angela Amoresano, and et al. 2024. "Analytical Investigation of the Profile of Human Chorionic Gonadotropin in Highly Purified Human Menopausal Gonadotrophin Preparations" International Journal of Molecular Sciences 25, no. 17: 9405. https://doi.org/10.3390/ijms25179405
APA StyleCapolupo, A., Petrocchi, S., Melchiorre, M., Jonas, K., D’Hooghe, T., Hanyaloglu, A., Sunkara, S., Palmese, A., Ozgumus, B., Amoresano, A., Angiuoni, G., Montenegro, S., Simone, P., & Lispi, M. (2024). Analytical Investigation of the Profile of Human Chorionic Gonadotropin in Highly Purified Human Menopausal Gonadotrophin Preparations. International Journal of Molecular Sciences, 25(17), 9405. https://doi.org/10.3390/ijms25179405