The Influence of Different Culture Media on the Growth and Recombinant Protein Production of Iranian Lizard Leishmania Promastigote
Abstract
Background: Leishmania is a eukaryotic protozoan parasite belonging to the Trypanosomatidae family. The Iranian Lizard Leishmania (I.L.L.), which is non-pathogenic to mammals, shows great promise to be used as an expression system for recombinant protein production. Unlike other Leishmania strains, the ideal culture medium for I.L.L. has not been established, although it is commonly cultured in the RPMI1640 medium.
Methods: We investigated the growth rate of the wild and recombinant I.L.L. in BHI, RPMI1640, LB, and M199 media with and without FBS, hemin, or lyophilized rabbit serum. Subsequently, the expression rate of the recombinant protein in these media was compared.
Results: The growth rate of I.L.L. in RPMI1640 medium and LB broth was similar and supplementation with 10% FBS did not affect the growth rate. The amount of protein expression in the LB medium was higher than in the other three media.
Conclusion: The LB broth is an appropriate medium for I.L.L. culture and recombinant protein production.
1. Klatt S, Simpson L, Maslov DA, Konthur Z. Leishmania tarentolae: Taxonomic classification and its application as a promising biotechnological expression host. PLoS Negl Trop Dis, 2019;13:e0007424-e0007424.
2. Fritsche C, Sitz M, Weiland N, Breitling R, Pohl HD. Characterization of the growth behavior of Leishmania tarentolae: A new expression system for recombinant proteins. J Basic Microbiol. 2007;47:384-393.
3. Basile G, Peticca M. Recombinant protein expression in Leishmania tarentolae. Mol Biotechnol. 2009;43:273.
4. Donyavi T, Bandehpour M, Kazemi B. Preparation of transgenic iranian lizard Leishmania coding hil-12.. Iran J Microbiol. 2017;9:305-311.
5. Mirzaahmadi S, Asaadi-Tehrani G, Bandehpour M, et al. Expression of recombinant human coagulation factor vii by the lizard Leishmania expression system. J Biomed Biotechnol. 2011;2011:873874.
6. Taromchi AH, Kazemi B, Mahmazi S, Bandehpour M. Heterologous expression of human il-29 (ifn-î»1) in iranian lizard Leishmania. Iran J Biotech. 2013;11:168-174.
7. Yadegari Z, Bandehpour M, Kazemi B, Sharifi-Sarasiabi K. Expression of recombinant human amelogenin in Iranian lizard Leishmania and its biological function assay. Iran J Public Health. 2015;44:987-996.
8. Handman E. Cell biology of Leishmania. Adv Parasitol. 1999;44:1-39.
9. Zhang W-W, Lypaczewski P, Matlashewski G. Application of crispr/cas9-mediated genome editing in Leishmania. In: Michels PAM, Ginger ML, Zilberstein D, eds. Trypanosomatids: Methods and protocols. New York, NY: Springer US; 2020:199-224.
10. Siripattanapipong S, Boontanom P, Leelayoova S, Mungthin M, Tan-ariya P. In vitro growth characteristics and morphological differentiation of Leishmania martiniquensis promastigotes in different culture media. Acta Trop. 2019;197:105039.
11. Rezaei Z, Van Reet N, Pouladfar G, et al. Expression of a rk39 homologue from an iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis. Parasites & Vectors. 2019;12:593.
12. Nasiri V. Sheep blood-lb agar base medium (slm) as a simple and suitable medium for the cultivation of Leishmania major promastigotes. Parasitol Res. 2013;112:3741-3742.
13. Grillberger L, Kreil TR, Nasr S, Reiter M. Emerging trends in plasma-free manufacturing of recombinant protein therapeutics expressed in mammalian cells. Biotechnol J. 2009;4:186-201.
14. Wingfield PT. Overview of the purification of recombinant proteins. Curr Protoc Protein Sci 2015;80:6.1.1-6.1.35.
15. Taheri T, Seyed N, Mizbani A, Rafati S. Leishmania-based expression systems. Appl Microbiol Biotechnol. 2016;100:7377-7385.
16. Meehan HA, Lundberg RA, Connell GJ. A trypanosomatid protein specifically interacts with a mammalian iron-responsive element. Parasitol Res. 2000;86:109-114.
17. Nasiri V, Dalimi A, Ghaffarifar F. Lb broth-lyophilized rabbit serum (llr) as a new and suitable culture medium for cultivation of promastigotes of Leishmania major. J Parasit Dis. 2017;41:247-251.
18. Kazemi B , Tahvildar-Bideroni GH, Hashemi Feshareki SR, Javadian E. Isolation a Lizard Leishmania promastigote from its Natural Host in Iran. J Biol Sci. 2004; 4: 620-623.
19. Jalali A, Bandehpour M, Chegeni R, et al. Expression, purification, and evaluation of anti-il-2rα antibody secreted by Leishmania tarentolae. International Journal of Peptide Research and Therapeutics. 2021;27:301-307.
20. Cunha J, Carrillo E, Sánchez C, Cruz I, Moreno J, Cordeiro-da-Silva A. Characterization of the biology and infectivity of Leishmania infantum viscerotropic and dermotropic strains isolated from hiv+ and hiv- patients in the murine model of visceral leishmaniasis. Parasites & Vectors. 2013;6:122.
21. BM Jalali Akram, B Kazemi. Compound serum and hemin free medium for cultivation of Leishmania tarentolae: A recombinant protein expression system. Afr J Biotechnol. 2015;14(25):2130-2134. https://doi.org/10.5897/AJB2015.14654.
22. Gupta V, Sengupta M, Prakash J, Tripathy BC. Production of recombinant pharmaceutical proteins. Basic and Applied Aspects of Biotechnology. 2016:77-101.
23. Khow O, Suntrarachun S. Strategies for production of active eukaryotic proteins in bacterial expression system. Asian Pac J Trop Biomed. 2012;2:159-162.
24. Legastelois I, Buffin S, Peubez I, Mignon C, Sodoyer R, Werle B. Non-conventional expression systems for the production of vaccine proteins and immunotherapeutic molecules. Hum Vaccin Immunother. 2017;13:947-961.
25. Niimi T. Recombinant protein production in the eukaryotic protozoan parasite Leishmania tarentolae: A review. Methods Mol Biol. 2012;824:307-315.
2. Fritsche C, Sitz M, Weiland N, Breitling R, Pohl HD. Characterization of the growth behavior of Leishmania tarentolae: A new expression system for recombinant proteins. J Basic Microbiol. 2007;47:384-393.
3. Basile G, Peticca M. Recombinant protein expression in Leishmania tarentolae. Mol Biotechnol. 2009;43:273.
4. Donyavi T, Bandehpour M, Kazemi B. Preparation of transgenic iranian lizard Leishmania coding hil-12.. Iran J Microbiol. 2017;9:305-311.
5. Mirzaahmadi S, Asaadi-Tehrani G, Bandehpour M, et al. Expression of recombinant human coagulation factor vii by the lizard Leishmania expression system. J Biomed Biotechnol. 2011;2011:873874.
6. Taromchi AH, Kazemi B, Mahmazi S, Bandehpour M. Heterologous expression of human il-29 (ifn-î»1) in iranian lizard Leishmania. Iran J Biotech. 2013;11:168-174.
7. Yadegari Z, Bandehpour M, Kazemi B, Sharifi-Sarasiabi K. Expression of recombinant human amelogenin in Iranian lizard Leishmania and its biological function assay. Iran J Public Health. 2015;44:987-996.
8. Handman E. Cell biology of Leishmania. Adv Parasitol. 1999;44:1-39.
9. Zhang W-W, Lypaczewski P, Matlashewski G. Application of crispr/cas9-mediated genome editing in Leishmania. In: Michels PAM, Ginger ML, Zilberstein D, eds. Trypanosomatids: Methods and protocols. New York, NY: Springer US; 2020:199-224.
10. Siripattanapipong S, Boontanom P, Leelayoova S, Mungthin M, Tan-ariya P. In vitro growth characteristics and morphological differentiation of Leishmania martiniquensis promastigotes in different culture media. Acta Trop. 2019;197:105039.
11. Rezaei Z, Van Reet N, Pouladfar G, et al. Expression of a rk39 homologue from an iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis. Parasites & Vectors. 2019;12:593.
12. Nasiri V. Sheep blood-lb agar base medium (slm) as a simple and suitable medium for the cultivation of Leishmania major promastigotes. Parasitol Res. 2013;112:3741-3742.
13. Grillberger L, Kreil TR, Nasr S, Reiter M. Emerging trends in plasma-free manufacturing of recombinant protein therapeutics expressed in mammalian cells. Biotechnol J. 2009;4:186-201.
14. Wingfield PT. Overview of the purification of recombinant proteins. Curr Protoc Protein Sci 2015;80:6.1.1-6.1.35.
15. Taheri T, Seyed N, Mizbani A, Rafati S. Leishmania-based expression systems. Appl Microbiol Biotechnol. 2016;100:7377-7385.
16. Meehan HA, Lundberg RA, Connell GJ. A trypanosomatid protein specifically interacts with a mammalian iron-responsive element. Parasitol Res. 2000;86:109-114.
17. Nasiri V, Dalimi A, Ghaffarifar F. Lb broth-lyophilized rabbit serum (llr) as a new and suitable culture medium for cultivation of promastigotes of Leishmania major. J Parasit Dis. 2017;41:247-251.
18. Kazemi B , Tahvildar-Bideroni GH, Hashemi Feshareki SR, Javadian E. Isolation a Lizard Leishmania promastigote from its Natural Host in Iran. J Biol Sci. 2004; 4: 620-623.
19. Jalali A, Bandehpour M, Chegeni R, et al. Expression, purification, and evaluation of anti-il-2rα antibody secreted by Leishmania tarentolae. International Journal of Peptide Research and Therapeutics. 2021;27:301-307.
20. Cunha J, Carrillo E, Sánchez C, Cruz I, Moreno J, Cordeiro-da-Silva A. Characterization of the biology and infectivity of Leishmania infantum viscerotropic and dermotropic strains isolated from hiv+ and hiv- patients in the murine model of visceral leishmaniasis. Parasites & Vectors. 2013;6:122.
21. BM Jalali Akram, B Kazemi. Compound serum and hemin free medium for cultivation of Leishmania tarentolae: A recombinant protein expression system. Afr J Biotechnol. 2015;14(25):2130-2134. https://doi.org/10.5897/AJB2015.14654.
22. Gupta V, Sengupta M, Prakash J, Tripathy BC. Production of recombinant pharmaceutical proteins. Basic and Applied Aspects of Biotechnology. 2016:77-101.
23. Khow O, Suntrarachun S. Strategies for production of active eukaryotic proteins in bacterial expression system. Asian Pac J Trop Biomed. 2012;2:159-162.
24. Legastelois I, Buffin S, Peubez I, Mignon C, Sodoyer R, Werle B. Non-conventional expression systems for the production of vaccine proteins and immunotherapeutic molecules. Hum Vaccin Immunother. 2017;13:947-961.
25. Niimi T. Recombinant protein production in the eukaryotic protozoan parasite Leishmania tarentolae: A review. Methods Mol Biol. 2012;824:307-315.
| Files | ||
| Issue | Vol 17 No 4 (2022) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v17i4.11282 | |
| Keywords | ||
| Leishmania Luria–Bertani M199 RPMI1640 Brain heart infusion Lyophilized rabbit serum | ||
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How to Cite
1.
Abdi Ghavidel A, Aghamiri S, Jajarmi V, Bandehpour M, Kazemi B. The Influence of Different Culture Media on the Growth and Recombinant Protein Production of Iranian Lizard Leishmania Promastigote. Iran J Parasitol. 2022;17(4):543-553.
