Search: onr:"swepub:oai:gup.ub.gu.se/106560" >
Recombinant MUC1 mu...
Recombinant MUC1 mucin with a breast cancer-like O-glycosylation produced in large amounts in Chinese-hamster ovary cells.
-
- Bäckström, Malin, 1967 (author)
- Gothenburg University,Göteborgs universitet,Institutionen för medicinsk och fysiologisk kemi,Institute of Medical Biochemistry
-
Link, Thomas (author)
-
- Olson, Fredrik J., 1975 (author)
- Gothenburg University,Göteborgs universitet,Institutionen för medicinsk och fysiologisk kemi,Institute of Medical Biochemistry
-
show more...
-
- Karlsson, Hasse, 1943 (author)
- Gothenburg University,Göteborgs universitet,Institutionen för medicinsk och fysiologisk kemi,Institute of Medical Biochemistry
-
Graham, Rosalind (author)
-
Picco, Gianfranco (author)
-
Burchell, Joy (author)
-
Taylor-Papadimitriou, Joyce (author)
-
Noll, Thomas (author)
-
- Hansson, Gunnar C., 1951 (author)
- Gothenburg University,Göteborgs universitet,Institutionen för medicinsk och fysiologisk kemi,Institute of Medical Biochemistry
-
show less...
-
(creator_code:org_t)
- 2003
- 2003
- English.
-
In: The Biochemical journal. - 1470-8728. ; 376:Pt 3, s. 677-86
- Related links:
-
https://gup.ub.gu.se...
-
show more...
-
https://doi.org/10.1...
-
show less...
Abstract
Subject headings
Close
- We have developed an expression system for the production of large quantities of recombinant MUC1 mucin in CHO-K1 (Chinese-hamster ovary K1) cells. The extracellular part of human MUC1, including 16 MUC1 tandem repeats, was produced as a fusion protein with murine IgG Fc, with an intervening enterokinase cleavage site for the removal of the Fc tail. Stable MUC1-IgG-producing CHO-K1 clones were generated and were found to secrete MUC1-IgG into the culture medium. After adaptation to suspension culture in protein-free medium in a bioreactor, the fusion protein was secreted in large quantities (100 mg/l per day) into the culture supernatant. From there, MUC1 could be purified to homogeneity using a two-step procedure including enterokinase cleavage and ion-exchange chromatography. Capillary liquid chromatography MS of released oligosaccharides from CHO-K1-produced MUC1 identified the main O-glycans as Galbeta1-3GalNAc (core 1) and mono- and di-sialylated core 1. The glycans occupied on average 4.3 of the five potential O-glycosylation sites in the tandem repeats, as determined by nano-liquid chromatography MS of partially deglycosylated Clostripain-digested protein. A very similar O-glycan profile and site occupancy was found in MUC1-IgG produced in the breast carcinoma cell line T47D, which has O-glycosylation typical for breast cancer. In contrast, MUC1-IgG produced in another breast cancer cell line, MCF-7, showed a more complex pattern with both core 1- and core 2-based O-glycans. This is the first reported production of large quantities of recombinant MUC1 with a breast cancer-like O-glycosylation that could be used for the immunotherapy of breast cancer.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
Keyword
- Amino Acids
- analysis
- Animals
- Antibodies
- Monoclonal
- immunology
- Breast Neoplasms
- metabolism
- CHO Cells
- Carbohydrate Sequence
- Carcinoma
- metabolism
- Cell Line
- Tumor
- Cricetinae
- Female
- Glycosylation
- Humans
- Immunoglobulin G
- genetics
- Molecular Sequence Data
- Mucin-1
- chemistry
- genetics
- metabolism
- Polysaccharides
- chemistry
- Recombinant Fusion Proteins
- chemistry
- isolation & purification
- metabolism
Publication and Content Type
- ref (subject category)
- art (subject category)
Find in a library
To the university's database
- By the author/editor
-
Bäckström, Malin ...
-
Link, Thomas
-
Olson, Fredrik J ...
-
Karlsson, Hasse, ...
-
Graham, Rosalind
-
Picco, Gianfranc ...
-
show more...
-
Burchell, Joy
-
Taylor-Papadimit ...
-
Noll, Thomas
-
Hansson, Gunnar ...
-
show less...
- About the subject
-
- MEDICAL AND HEALTH SCIENCES
-
MEDICAL AND HEAL ...
-
and Medical Biotechn ...
-
and Medical Biotechn ...
- Articles in the publication
-
The Biochemical ...
- By the university
-
University of Gothenburg