For example, various N-glycans were transferred from the corresponding oxazolines to the GlcNAc residue on the intact anti-Her2 antibody with a mutant endoglycosidase, affording glyco-engineered anti-Her2 antibodies (Fig.
developed an endoglycosidase having a lower hydrolysis activity from Endo-S by using the same strategy as that for the creation of Endo-M mutant.
H (Endo H) was purchased from New England Biolabs (Beverly, MA) carboxy methyl cellulose (CMC) was purchased from Sigma (St.
Studies from our laboratory also revealed the majority of breast cancer cell lines harbor differential glycosylation patterns (specifically in the secreted pro-CatD), with prevalence of endoglycosidase
H (EndoH) sensitive N-glycan structures (Figure 2 B and 2C, Khalkhali-Ellis, unpublished observations) .
Hyaluronidase is an endoglycosidase
that degrades hyaluronic acid into smaller fragments.
The cancer cells release an endoglycosidase
(N-acetylgalactosaminidase) that inactivates Gc-MAF.
The PLB, both the native and modified forms, were incubated with endoglycosidase H for 180 min.
These PLBs contained about 50% carbohydrate and the soluble PLB was digested with endoglycosidase H for 24 h and liberated about 75% of the total carbohydrate.
In contrast, O-glycans are built on different protein glycan linkages and have extremely diverse structures; in addition, there is no endoglycosidase
available for the release of intact O-glycans.
The difference between these values and the value of 70 kDa estimated from SDSPAGE of cerebral muscarinic receptors was supposed to be accounted for by the contribution of the carbohydrate moiety, and was confirmed by endoglycosidase
treatment of cerebral and atrial receptors.
The construct FH18-20 was found to run as a doublet of bands in SDS-PAGE, and the upper band was verified as a glycosylated form by treatment with endoglycosidase
H (Sigma; data not shown).
F2, AND PEPTIDE: N-GLYCOSIDASE F TREATMENTS