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Affinity-matured re...
Affinity-matured recombinant immunotoxin targeting gangliosides 3 '-isoLM1 and 3 ', 6 '-isoLD1 on malignant gliomas
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Piao, H. L. (författare)
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Kuan, C. T. (författare)
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Chandramohan, V. (författare)
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Keir, S. T. (författare)
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Pegram, C. N. (författare)
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Bao, X. H. (författare)
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- Månsson, Jan-Eric, 1946 (författare)
- Gothenburg University,Göteborgs universitet,Institutionen för neurovetenskap och fysiologi, sektionen för psykiatri och neurokemi,Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry
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Pastan, I. H. (författare)
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Bigner, D. D. (författare)
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(creator_code:org_t)
- 2014-10-27
- 2013
- Engelska.
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Ingår i: Mabs. - : Informa UK Limited. - 1942-0862 .- 1942-0870. ; 5:5, s. 748-762
- Relaterad länk:
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https://www.tandfonl...
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https://gup.ub.gu.se...
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https://doi.org/10.4...
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Abstract
Ämnesord
Stäng
- About 60 percent of glioblastomas highly express the gangliosides 3-isoLM1 and 3,6-isoLD1 on the cell surface, providing ideal targets for brain tumor immunotherapy. A novel recombinant immunotoxin, DmAb14m-(scFv)-PE38KDEL (DmAb14m-IT), specific for the gangliosides 3-isoLM1 and 3,6-isoLD1, was constructed with improved affinity and increased cytotoxicity for immunotherapeutic targeting of glioblastoma. We isolated an scFv parental clone from a previously established murine hybridoma, DmAb14, that is specific to both 3-isoLM1 and 3,6-isoLD1. We then performed in vitro affinity maturation by CDR hotspot random mutagenesis. The binding affinity and specificity of affinity-matured DmAb14m-IT were measured by surface-plasmon resonance, flow cytometry, and immunohistochemical analysis. In vitro cytotoxicity of DmAb14m-IT was measured by protein synthesis inhibition and cell death assays in human cell lines expressing gangliosides 3-isoLM1 and 3,6-isoLD1 (D54MG and D336MG) and xenograft-derived cells (D2224MG). As a result, the K-D of DmAb14m-IT for gangliosides 3-isoLM1 and 3,6-isoLD1 was 2.6 x 10(-9)M. Also, DmAb14m-IT showed a significantly higher internalization rate in cells expressing 3-isoLM1 and 3,6-isoLD1. The DmAb14m-IT IC50 was 80 ng/mL (1194 pM) on the D54MG cell line, 5 ng/ml (75 pM) on the D336MG cell line, and 0.5 ng/ml (7.5 pM) on the D2224MG xenograft-derived cells. There was no cytotoxicity on ganglioside-negative HEK293 cells. Immunohistochemical analysis confirmed the specific apparent affinity of DmAb14m-IT with 3-isoLM1 and 3,6-isoLD1. In conclusion, DmAb14m-IT showed specific binding affinity, a significantly high internalization rate, and selective cytotoxicity on glioma cell lines and xenograft-derived cells expressing 3-isoLM1 and 3,6-isoLD1, thereby displaying robust therapeutic potential for testing the antitumor efficacy of DmAb14m-IT at the preclinical level and eventually in the clinical setting.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Clinical Medicine (hsv//eng)
Nyckelord
- ganglioside
- recombinant immunotoxin
- single-chain variable fragment
- glioblastoma
- affinity
- SINGLE-CHAIN IMMUNOTOXIN
- IN-VITRO
- MONOCLONAL-ANTIBODIES
- TUMORS
- BRAIN
- EXPRESSION
- GROWTH
- GD3
- MENINGITIS
- XENOGRAFTS
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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