碩士 / 國立中興大學 / 生命科學院碩士在職專班 / 102 / Glycoengineered therapeutic antibodies lacking core fucose residues exhibit strong ADCC, in the same time, the high binding with the Fc gamma recepter IIIa on the effector cell surface evade the serum IgG inhibitory effect. We aimed to use enzymatic modification of N-glycans to produce non-fucosylated forms on human antibodies.
In this study, we searched for α-L-fucosidase candidates from Lactobacillus, Bifidobacterium and Bacteroides species. To confirm fucosidase activities, several methods was established: (1) Lectin blot with the use of Aleuria aurantia Lectin; (2) in vitro enzymatic analysis with 4-nitrophenyl fucopyranoside; (3) N-glycan profiling of antibodies. The results showed that (His)6-tagged recombinant α-L-fucosidase AlfC from the Lactobacillus casei strain BL23, combined with endoglycosidases, has the activity to release fucose residues on human IgG and other glycoproteins.
Further, we use surface display techniques to express AlfC on the surface of E. coli cells. In the end, we identified a suitable system 183 for surface display of AlfC.
| Identifer | oai:union.ndltd.org:TW/102NCHU5105053 |
| Date | January 2014 |
| Creators | Pei-Jung Yang, 楊佩蓉 |
| Contributors | Pin-Ju Chueh, 闕斌如 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 33 |
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