Efficient long-term and high-yielded production of a recombinant proteoglycan in eukaryotic HEK293 cells using a membrane-based bioreactor

Vascular endocan (ESM‐1) is markedly overexpressed in clear cell renal cell carcinoma

“…Endocan was originally described as a soluble proteoglycan secreted by endothelial cells 7–24 . Intratumour overexpression of endocan mRNA was previously recognized as being a molecular signature, indicating a bad prognosis, of highly vascularized breast, lung and melanoma tumours 9–11 .…”

“…Endocan was originally described as a soluble proteoglycan secreted by endothelial cells. [7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24] Intratumour overexpression of endocan mRNA was previously recognized as being a molecular signature, indicating a bad prognosis, of highly vascularized breast, lung and melanoma tumours. [9][10][11] More recently, intratumour overexpression of endocan protein has been evidenced by immunohistochemistry in the lung, and in hepatocarcinoma.…”

“…Human endocan was detected and quantified in sera from patients and healthy volunteers by using a sandwich ELISA, as previously described. 20,23 Briefly, a capture anti-endocan monoclonal antibody (clone MEP 14, U774) was directly adsorbed overnight at 4°C onto microwells (Maxisorp plate; NUNC, Roskilde, Denmark). After washing, the plates were blocked with 10% fetal bovine serum (FBS) in phosphate-buffered saline for 1 h at room temperature.…”

Recombinant production of proteoglycans and their bioactive domains

“…The serum‐borne PG, endocan, which is highly expressed in the vasculature around tumours, has been expressed in HEK‐293 cells and shown to have a relatively longer CS/DS chain than the endocan purified from endothelial cells. Given this, they had a similar disaccharide composition suggesting that their microstructures, which are responsible for binding growth factors and cytokines, are very similar .…”

Avaliação do potencial de crescimento e produção de proteínas recombinantes de células humanas adaptadas para crescimento em suspensão e meios de cultura livres de soro fetal bovino

“…A célula HEK293 é uma linhagem derivada de células embrionárias humanas de rim transformada com os genes do adenovirus E1A e E1B (GRAHAM et al, 1977). Esta linhagem celular humana é provavelmente a mais utilizada para pesquisas relacionadas à expressão de proteínas recombinantes devido ao seu crescimento em suspensão e em meios livres de soro, a facilidade para transfecção transiente através de diferentes métodos e alta produtividade, como descrito em alguns trabalhos na literatura (SUN et al, 2006;ADAM et al, 2008;BACKLIWAL et al, 2008;LOIGNON et al, 2008;WAJIH et al, 2008;SONG et al, 2011;DUMONT et al, 2012;FISCHER et al, 2012, HACKER et al, 2013 KUNERT e REINHART, 2016). Por esses motivos, também é amplamente utilizada para a produção de vetores virais, como adenovírus, retrovírus, lentivírus e vírus adenoassociado, muitos em fase II e III de ensaios clínicos (KIM et al, 2009;HENRY et al, 2011;PETIOT et al, 2011).…”