[논문]Selective endocytosis of recombinant human BMPs through cell surface heparan sulfate proteoglycans in CHO cells: BMP-2 and BMP-7

Kim, Mi Gyeom; Kim, Che Lin; Kim, Young Sik; Jang, Ju Woong; Lee, Gyun Min

doi:10.1038/s41598-021-82955-1

[해외논문] Selective endocytosis of recombinant human BMPs through cell surface heparan sulfate proteoglycans in CHO cells: BMP-2 and BMP-7 원문보기

Scientific reports, v.11 no.1, 2021년, pp.3378 -

Kim, Mi Gyeom (Department of Biological Sciences, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 34141 Republic of Korea) , Kim, Che Lin (The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Lyngby, Denmark) , Kim, Young Sik (Institute of Biomaterial and Medical Engineering, Cellumed, Seoul, Republic of Korea) , Jang, Ju Woong (Institute of Biomaterial and Medical Engineering, Cellumed, Seoul, Republic of Korea) , Lee, Gyun Min (Department of Biological Sciences, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 34141 Republic of Korea)

Abstract ▼ AI-Helper

Cell surface heparan sulfate proteoglycan (HSPG)-mediated endocytosis results in poor yields of recombinant human bone morphogenetic proteins (rhBMPs) from CHO cell cultures. Upon incubation of rhBMP-2 and rhBMP-7 with CHO cells at 37 °C, both rhBMP-2 and rhBMP-7 bound to the cell surface HSPGs in CHO cells, but only rhBMP-2 was actively internalized into CHO cells. Cell surface HSPGs were found to serve as the main receptor for rhBMP-2 internalization. It was also found that the cell surface HSPG-mediated endocytosis of rhBMP-2 occurred through both the clathrin- and caveolin-dependent pathways. Blockage of rhBMP-2 internalization by the addition of structural analogs of HSPGs such as dextran sulfate (DS) and heparin dramatically increased rhBMP-2 production in recombinant CHO (rCHO) cell cultures. Compared to the control cultures, addition of DS (1.0 g/L) and heparin (0.2 g/L) resulted in a 22.0- and 19.0-fold increase in the maximum rhBMP-2 concentration, respectively. In contrast, the production of rhBMP-7, which was not internalized into the rCHO cells, did not dramatically increase upon addition of DS and heparin. Taken together, rhBMPs have a different fate in terms of HSPG-mediated internalization in CHO cells. HSPG-mediated endocytosis of each rhBMP should be understood individually to increase the rhBMP yield in rCHO cell cultures.

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