Selection of novel peptide mimics of the GD2 ganglioside from a constrained phage-displayed peptide library
Aberrant glycosylation is a universal feature of cancer cells. There are quantitative and qualitative changes in expression of gangliosides observed in tumors of a neuroectodermal origin such as neuroblastoma, melanoma and astrocytoma. The presence of large amounts of GD2 ganglioside on neuroblastom...
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Published in | International journal of molecular medicine Vol. 19; no. 5; pp. 829 - 839 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
Greece
D.A. Spandidos
01.05.2007
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Subjects | |
Online Access | Get full text |
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Summary: | Aberrant glycosylation is a universal feature of cancer cells. There are
quantitative and qualitative changes in expression of gangliosides observed in
tumors of a neuroectodermal origin such as neuroblastoma, melanoma and astrocytoma.
The presence of large amounts of GD2 ganglioside on neuroblastoma cells, as compared
to normal cells, opens the possibilities to use the tumor-associated carbohydrate
antigen in diagnosis and immunotherapeutic approaches. In the quest for immunogens
potentially capable of eliciting anti-GD2 ganglioside immune responses, we performed
affinity purification of phage-displayed peptides from the LX-8 library (12-mer
containing disulphide bridge). The library was screened with the biotinylated
anti-GD2 ganglioside 14G2a mAb monoclonal antibody. Our goal was to isolate and
characterize peptide mimics of GD2 ganglioside. Numerous individual phage clones
that bound 14G2a mAb were identified with the application of immunoblotting technique
in the phage pools yielded from the pannings. The phage-borne peptides were tested
for their anti-GD2 ganglioside antibody binding ability using ELISA. Among these
clones five different phage-displayed peptide sequences were identified. Moreover,
we showed that the secondary structure of the peptides, stabilized by the disulfide
bridging between cysteine residues at positions 2 and 11, was crucial for the
binding of the peptides to 14G2a mAb. In a separate set of experiments, we observed
a competition of the peptides, expressed on phages as well as in their synthetic
form, with the nominal antigen GD2 ganglioside expressed on IMR-32 neuroblastoma
cells for binding to 14G2a mAb. Based on the obtained results we concluded that
all of these 5 peptides were mimics of the GD2 ganglioside. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 1107-3756 1791-244X |
DOI: | 10.3892/ijmm.19.5.829 |