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Theriogenology2021; 172; 8-19; doi: 10.1016/j.theriogenology.2021.05.013

Development of a suitable manufacturing process for production of a bioactive recombinant equine chorionic gonadotropin (reCG) in CHO-K1 cells.

Abstract: Equine chorionic gonadotropin (eCG) is a heterodimeric glycoprotein hormone produced by pregnant mares that has been used to improve reproductive performance in different domestic species. Several strategies to produce the hormone in a recombinant way have been reported; nevertheless, no approach has been able to produce a recombinant eCG (reCG) with significant in vivo bioactivity or in sufficient quantities for commercial purposes. For this reason, the only current product available on the market consists of partially purified preparations from serum of pregnant mares (PMSG). Herein, we describe a highly efficient process based on third-generation lentiviral vectors as delivery method for the production of reCG in suspension CHO-K1 cells, with productivities above 20 IU 10 cell.d and 70% purification yields after one purification step. Importantly, reCG demonstrated biological activity in cattle, since around 30 μg of reCG were needed to exert the same biologic effect of 400 IU of PMSG in an ovulation synchronization protocol. The results obtained demonstrate that the developed strategy represents an attractive option for the production of reCG and constitutes an auspicious alternative for the replacement of animals as a source of PMSG.
Copyright © 2021 Elsevier Inc. All rights reserved.
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Publication Date: 2021-05-23 PubMed ID: 34082223DOI: 10.1016/j.theriogenology.2021.05.013Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article discusses a new efficient manufacturing process for producing a bioactive recombinant equine chorionic gonadotropin (reCG) hormone in CHO-K1 cells, which shown significant in vivo bioactivity and may serve as an alternative for the currently used animal source.

Introduction

  • The paper focuses on the production of the equine chorionic gonadotropin (eCG) hormone. This hormone is produced by pregnant mares and has applications in improving reproductive performance in various domestic species.
  • While there have been several strategies to produce eCG recombinantly, previous methods have not been successful in generating a recombinant eCG (reCG) with significant in vivo bioactivity or in sufficient quantities for commercial use.
  • Currently, the only product available in the market is derived from the serum of pregnant mares (PMSG), a method which has ethical and supply restrictions.

Methods

  • The researchers developed and described a high-efficiency process that makes use of third-generation lentiviral vectors to produce reCG in suspension CHO-K1 cells.
  • The process showed productivity above 20 IU 10 cells per day and achieved 70% purification yields after just one purification step.

Results

  • The reCG produced showed significant biological activity in cattle. The study found that approximately 30 μg of reCG was needed to achieve the same biological effect as 400 IU of PMSG in an ovulation synchronization protocol.
  • The results demonstrate that the developed strategy is a viable option for the commercial production of reCG.

Conclusion

  • This manufacturing process potentially paves the way for using reCG as a replacement for animals as a source of PMSG, thus addressing ethical and supply concerns.

Cite This Article

APA
Villarraza CJ, Antuña S, Tardivo MB, Rodríguez MC, Mussio P, Cattaneo L, Fontana D, Díaz PU, Ortega HH, Tríbulo A, Macagno A, Bó GA, Ceaglio N, Prieto C. (2021). Development of a suitable manufacturing process for production of a bioactive recombinant equine chorionic gonadotropin (reCG) in CHO-K1 cells. Theriogenology, 172, 8-19. https://doi.org/10.1016/j.theriogenology.2021.05.013

Publication

Theriogenology
ISSN: 1879-3231
NlmUniqueID: 0421510
Country: United States
Language: English
Volume: 172
Pages: 8-19
PII: S0093-691X(21)00173-4

Researcher Affiliations

Villarraza, Carlos Javier
  • UNL, CONICET, FBCB (School of Biochemistry and Biological Sciences), CBL (Biotechnological Center of Litoral), Cell Culture Laboratory, Ciudad Universitaria, Ruta Nacional 168 - Km 472.4 - C.C. 242, (S3000ZAA), Santa Fe, Argentina.
Antuña, Sebastián
  • Biotecnofe S.A., PTLC, Ruta 168 Pje El Pozo, (CP3000), Santa Fe, Argentina.
Tardivo, María Belén
  • Biotecnofe S.A., PTLC, Ruta 168 Pje El Pozo, (CP3000), Santa Fe, Argentina.
Rodríguez, María Celeste
  • UNL, CONICET, FBCB (School of Biochemistry and Biological Sciences), CBL (Biotechnological Center of Litoral), Cell Culture Laboratory, Ciudad Universitaria, Ruta Nacional 168 - Km 472.4 - C.C. 242, (S3000ZAA), Santa Fe, Argentina.
Mussio, Pablo
  • UNL, FBCB (School of Biochemistry and Biological Sciences), CBL (Biotechnological Center of Litoral), Biotechnological Development Laboratory, Ciudad Universitaria, Ruta Nacional 168 - Km 472.4 - C.C. 242 - (S3000ZAA), Santa Fe, Argentina.
Cattaneo, Luciano
  • UNL, FCV, R.P. Kreder, 2805, Esperanza, Santa Fe, Argentina.
Fontana, Diego
  • UNL, CONICET, FBCB (School of Biochemistry and Biological Sciences), CBL (Biotechnological Center of Litoral), Cell Culture Laboratory, Ciudad Universitaria, Ruta Nacional 168 - Km 472.4 - C.C. 242, (S3000ZAA), Santa Fe, Argentina; Cellargen Biotech S.R.L., Antonia Godoy 6369 (S3000ZAA), Santa Fe, Argentina; Biotecnofe S.A., PTLC, Ruta 168 Pje El Pozo, (CP3000), Santa Fe, Argentina.
Díaz, Pablo U
  • UNL, FCV, R.P. Kreder, 2805, Esperanza, Santa Fe, Argentina; Laboratorio de Biología Celular y Molecular Aplicada, Instituto de Ciencias Veterinarias Del Litoral (ICiVet Litoral), Universidad Nacional Del Litoral (UNL) / Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Esperanza, Santa Fe, Argentina.
Ortega, Hugo H
  • UNL, FCV, R.P. Kreder, 2805, Esperanza, Santa Fe, Argentina; Laboratorio de Biología Celular y Molecular Aplicada, Instituto de Ciencias Veterinarias Del Litoral (ICiVet Litoral), Universidad Nacional Del Litoral (UNL) / Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Esperanza, Santa Fe, Argentina.
Tríbulo, Andres
  • Instituto de Reproducción Animal Córdoba (IRAC), Paraje Pozo Del Tigre, Zona Rural Gral, Paz, (X5145), Córdoba, Argentina.
Macagno, Alejandro
  • Instituto AP de Ciencias Básicas y Aplicadas, Universidad Nacional de Villa María (UNVM), Villa Del Rosario, (X5963), Córdoba, Argentina.
Bó, Gabriel A
  • Instituto AP de Ciencias Básicas y Aplicadas, Universidad Nacional de Villa María (UNVM), Villa Del Rosario, (X5963), Córdoba, Argentina; Instituto de Reproducción Animal Córdoba (IRAC), Paraje Pozo Del Tigre, Zona Rural Gral, Paz, (X5145), Córdoba, Argentina.
Ceaglio, Natalia
  • UNL, CONICET, FBCB (School of Biochemistry and Biological Sciences), CBL (Biotechnological Center of Litoral), Cell Culture Laboratory, Ciudad Universitaria, Ruta Nacional 168 - Km 472.4 - C.C. 242, (S3000ZAA), Santa Fe, Argentina.
Prieto, Claudio
  • UNL, FBCB (School of Biochemistry and Biological Sciences), CBL (Biotechnological Center of Litoral), Biotechnological Development Laboratory, Ciudad Universitaria, Ruta Nacional 168 - Km 472.4 - C.C. 242 - (S3000ZAA), Santa Fe, Argentina; Cellargen Biotech S.R.L., Antonia Godoy 6369 (S3000ZAA), Santa Fe, Argentina; Biotecnofe S.A., PTLC, Ruta 168 Pje El Pozo, (CP3000), Santa Fe, Argentina. Electronic address: cprieto@fbcb.unl.edu.ar.

MeSH Terms

  • Animals
  • CHO Cells
  • Cattle
  • Chorionic Gonadotropin / pharmacology
  • Cricetinae
  • Cricetulus
  • Female
  • Gonadotropins, Equine / pharmacology
  • Horses
  • Ovulation
  • Pregnancy

Conflict of Interest Statement

Declaration of competing interest The authors declare no commercial or financial conflict of interest.

Citations

This article has been cited 7 times.
  1. Rodríguez MC, Mussio PE, Villarraza J, Tardivo MB, Antuña S, Fontana D, Ceaglio N, Prieto C. Physicochemical Characterization of a Recombinant eCG and Comparative Studies with PMSG Commercial Preparations. Protein J 2023 Feb;42(1):24-36.
    doi: 10.1007/s10930-023-10092-xpubmed: 36652139google scholar: lookup
  2. Sales JNS, Carvalho LR, Simões LMS, Lemos LA, Vicente MP, Silva RRR, Orlandi LO, Baruselli PS, de Souza JC. New perspectives and evolution of ovulation synchronization protocols in bovine females. Anim Reprod 2025;22(3):e20250048.
    doi: 10.1590/1984-3143-AR2025-0048pubmed: 40933866google scholar: lookup
  3. Byambaragchaa M, Park SH, Park MH, Kang MH, Min KS. Enhanced Production and Functional Characterization of Recombinant Equine Chorionic Gonadotropin (rec-eCG) in CHO-DG44 Cells. Biomolecules 2025 Feb 14;15(2).
    doi: 10.3390/biom15020289pubmed: 40001592google scholar: lookup
  4. Lösle M, Lin CW, Beil-Wagner J, Aebi M, Buch T. Comparison of pregnant mare serum gonadotropin products with surprising differences in protein content. Sci Rep 2025 Feb 25;15(1):6824.
    doi: 10.1038/s41598-025-90833-3pubmed: 40000800google scholar: lookup
  5. Baruselli PS, de Abreu LÂ, Catussi BLC, Oliveira ACDS, Rebeis LM, Gricio EA, Albertini S, Sales JNS, Rodrigues CA. Use of new recombinant proteins for ovarian stimulation in ruminants. Anim Reprod 2023;20(2):e20230092.
    doi: 10.1590/1984-3143-AR2023-0092pubmed: 37720727google scholar: lookup
  6. Bó GA, Menchaca A. Prohibition of hormones in animal reproduction: what to expect and what to do?. Anim Reprod 2023;20(2):e20230067.
    doi: 10.1590/1984-3143-AR2023-0067pubmed: 37720723google scholar: lookup
  7. Thompson RE, Meyers MA, Palmer J, Veeramachaneni DNR, Magee C, de Mestre AM, Antczak DF, Hollinshead FK. Production of Mare Chorionic Girdle Organoids That Secrete Equine Chorionic Gonadotropin. Int J Mol Sci 2023 May 31;24(11).
    doi: 10.3390/ijms24119538pubmed: 37298490google scholar: lookup
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