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Planta Med 2022; 88(12): 1060-1068
DOI: 10.1055/a-1677-4135
Biological and Pharmacological Activity
Original Papers

Establishment of the Carrot-Made LTB-Syn Antigen Cell Line in Shake Flask and Airlift Bioreactor Cultures

Christian Carreño-Campos
1   Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, México
,
Jaime I. Arevalo-Villalobos
2   Laboratorio de Biofarmacéuticos Recombinantes, Universidad Autónoma de San Luis Potosí, SLP, México
,
María Luisa Villarreal
1   Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, México
,
Anabel Ortiz-Caltempa
1   Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, México
,
Sergio Rosales-Mendoza
2   Laboratorio de Biofarmacéuticos Recombinantes, Universidad Autónoma de San Luis Potosí, SLP, México
› Author AffiliationsSupported by: Consejo Nacional de Ciencia y Tecnología 222714 to MLV
Supported by: Consejo Nacional de Ciencia y Tecnología 311879 to SRM
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Abstract

Carrot (Daucus carota) cells have been used to effectively manufacture recombinant biopharmaceuticals such as cytokines, vaccines, and antibodies. We generated the carrot cell line Z4, genetically modified to produce the LTB-Syn antigen, which is a fusion protein proposed for immunotherapy against synucleinopathies. In this work, the Z4 cell suspension line was cultivated to produce the LTB-Syn protein in a 250 mL shake flask and 2 L airlift bioreactor cultures grown for 45 and 30 days, respectively. Maximum biomass was obtained on day 15 in both the airlift bioreactor (35.00 ± 0.04 g/L DW) and shake flasks (17.00 ± 0.04 g/L DW). In the bioreactor, the highest LTB-Syn protein yield (1.52 ± 0.03 µg/g FW) was obtained on day 15; while the same occurred on day 18 for shake flasks (0.92 ± 0.02 µg/g FW). LTB-Syn protein levels were analyzed by GM1-ELISA and western blot. PCR analysis confirmed the presence of the transgene in the Z4 line. The obtained data demonstrate that the carrot Z4 cell suspension line grown in airlift bioreactors shows promise for a scale-up cultivation producing an oral LTB-Syn antigen.

Key words

Daucus carota - Apiaceae - LTB-Syn antigen - airlift bioreactor - suspension culture - synucleinopathies

Supporting Information

    A picture of the airlift bioreactor used in this study is provided as supporting information.

  • Supporting Information


Publication History

Received: 15 June 2021

Accepted after revision: 14 October 2021

Article published online:
16 December 2021

© 2021. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

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