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Recombinant MUC1 mu...
Recombinant MUC1 mucin with a breast cancer-like O-glycosylation produced in large amounts in Chinese-hamster ovary cells.
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- Bäckström, Malin, 1967 (författare)
- Gothenburg University,Göteborgs universitet,Institutionen för medicinsk och fysiologisk kemi,Institute of Medical Biochemistry
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Link, Thomas (författare)
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- Olson, Fredrik J., 1975 (författare)
- Gothenburg University,Göteborgs universitet,Institutionen för medicinsk och fysiologisk kemi,Institute of Medical Biochemistry
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- Karlsson, Hasse, 1943 (författare)
- Gothenburg University,Göteborgs universitet,Institutionen för medicinsk och fysiologisk kemi,Institute of Medical Biochemistry
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Graham, Rosalind (författare)
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Picco, Gianfranco (författare)
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Burchell, Joy (författare)
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Taylor-Papadimitriou, Joyce (författare)
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Noll, Thomas (författare)
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- Hansson, Gunnar C., 1951 (författare)
- Gothenburg University,Göteborgs universitet,Institutionen för medicinsk och fysiologisk kemi,Institute of Medical Biochemistry
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(creator_code:org_t)
- 2003
- 2003
- Engelska.
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Ingår i: The Biochemical journal. - 1470-8728. ; 376:Pt 3, s. 677-86
- Relaterad länk:
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https://gup.ub.gu.se...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- 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.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
Nyckelord
- 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
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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Till lärosätets databas
- Av författaren/redakt...
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Bäckström, Malin ...
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Link, Thomas
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Olson, Fredrik J ...
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Karlsson, Hasse, ...
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Graham, Rosalind
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Picco, Gianfranc ...
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visa fler...
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Burchell, Joy
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Taylor-Papadimit ...
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Noll, Thomas
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Hansson, Gunnar ...
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visa färre...
- Om ämnet
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- MEDICIN OCH HÄLSOVETENSKAP
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MEDICIN OCH HÄLS ...
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och Medicinsk biotek ...
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och Medicinsk biotek ...
- Artiklar i publikationen
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The Biochemical ...
- Av lärosätet
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Göteborgs universitet