Enhancement of human prolactin synthesis by sodium butyrate addition to serum-free CHO cell culture.
Sodium butyrate (NaBu) has been used as a productivity enhancer for the synthesis of recombinant proteins in Chinese hamster ovary (CHO) cells. Thus, the influence of NaBu on the production of recombinant human prolactin (hPRL) from CHO cells was investigated for the first time. CHO cell cultures were submitted to a treatment with different concentrations of NaBu (0.25 to 4 mM). Quantitative and qualitative analyses by reverse-phase high-performance liquid chromatography (RP-HPLC) and Western blot or SDS-PAGE, carried out directly on CHO-conditioned medium, showed that the highest hPRL expression was obtained with 1 mM NaBu. In vitro biological assays based on noble rat lymphoma (Nb2) and mouse pro-B lymphoma (Ba/F3-LLP) cells were carried out on purified hPRL. Its bioactivity in the presence of NaBu was not apparently different from that of the First International Reference Reagent of recombinant hPRL (WHO 97/714). Our results show that NaBu increased the synthesis of recombinant hPRL in CHO cells, apparently without compromising either its structure or function.
Rodrigues Goulart
Herbert
H
Biotechnology Department, IPEN-CNEN, Cidade Universitária, Avenida Professor Lineu Prestes 2242, São Paulo, SP, Brazil.
Arthuso
Fernanda dos Santos
Fdos S
Capone
Marcos Vinicius Nucci
MV
de Oliveira
Taís Lima
TL
Bartolini
Paolo
P
Soares
Carlos Roberto Jorge
CR
D000818:Animals; D001681:Biological Assay; D015153:Blotting, Western; D020148:Butyric Acid / Q000494:pharmacology*; D016466:CHO Cells; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D003412:Cricetulus; D016895:Culture Media, Serum-Free; D004247:DNA / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D011388:Prolactin / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification; D013997:Time Factors
D016895:Culture Media, Serum-Free; D011994:Recombinant Proteins; D020148:Butyric Acid; D011388:Prolactin; D004247:DNA