Overview
CHO Lec 3.2.8.1 cells have four independent mutations in the N- and O- glycosylation pathways (Stanley, 1989). N-linked carbohydrates produced by CHO Lec 3.2.8.1 cells are all of the high mannose type, but differ in the number of mannoses, ranging from Man9 to Man5. O-glycosylation is homogenous, with only a single GalNAc residue attached per site. When cultured in the presence of the alpha-glucosidase I inhibitor N-butyl-deoxynojirimycin (NB-DNJ), glycoproteins produced in CHO Lec 3.2.8.1 cells are almost completely susceptible to Endo H digestion (Davis, 1995; Ikemizu, 1999). Endo H cleaves chitobiose, leaving a single N-linked N-acetylglucosamine per site, which is ideal for maintenance of protein solubility and special carbohydrate-protein interactions, such as between the first N-acetyl glucosamine residue and tryptophan. (On intact proteins, Endo H cleaves (Man)5-9 chains but (Man)3-4 chains are resistant.)
Details
Details of this protocol, Establishment Of Stable Transfectant Of CHO Lec Cells, are located on a web site other than Biocompare Protocols.