Enhancement of Human Thyrotropin Synthesis by Sodium Butyrate Addition to Serum-Free CHO Cell Culture

Author(s)

Damiani, Renata ^[1] ; Almeida, Beatriz E. ^[1] ; Oliveira, Joao E. ^[1] ; Bartolini, Paolo ^[1] ; Ribela, Maria Teresa C. P. ^[1]

Affiliation

^[1] IPEN CNEN, Dept Biotechnol, BR-05508900 Sao Paulo - Brazil

• Document type: Journal article
• Source: Applied Biochemistry and Biotechnology; v. 171, n. 7, p. 1658-1672, DEC 2013.
• Web of Science Citations: 12

Abstract

The influence of sodium butyrate (NaBu) on the synthesis of recombinant human thyrotropin (r-hTSH) by CHO cells was investigated for the first time. A volumetric productivity of similar to 10 mu g hTSH/mL was repeatedly obtained, with a 3.3-fold increase over a control culture carried out in the absence of NaBu. Since NaBu can induce CHO cell apoptosis and cell growth arrest, the increase in specific productivity was even higher, i.e., ca. 5-fold. Analysis of the N-glycan composition of r-hTSH obtained with the addition of NaBu to the culture medium showed an approximately 12 % increase in the amount of sialic acid, as well as in total carbohydrate, partly due to the increase in the site occupancy from 2.77 to 2.93 glycans per mole of hTSH. The two hormone preparations were characterized by N-glycan structural analysis, which showed that NaBu increased the bi-antennary structures by ca. 13 % while decreasing the tri-antennary structures by approximately the same amount. The in vivo biological activity and pharmacokinetic behavior (clearance) were found to be similar for the two hormone preparations. (AU)

• FAPESP's process: 11/07289-0 - Synthesis and characterization of different glycoforms of recombinant human thyrotropin (rhTSH) pharmacologically actives
• Grantee: Maria Teresa de Carvalho Pinto Ribela
• Support Opportunities: Regular Research Grants