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. 2003 Sep;9(9):1920-4.
doi: 10.3748/wjg.v9.i9.1920.

Screening and identification of mimotope of gastric cancer associated antigen MGb1-Ag

Zhe-Yi Han  1 , Kai-Chun Wu, Feng-Tian He, Quan-Li Han, Yong-Zhan Nie, Ying Han, Xiao-Nan Liu, Jian-Yong Zheng, Mei-Hong Xu, Tao Lin, Dai-Ming Fan
Affiliations

Screening and identification of mimotope of gastric cancer associated antigen MGb1-Ag

Zhe-Yi Han et al. World J Gastroenterol. 2003 Sep.

Abstract

Aim: Using a monoclonal antibody against gastric cancer antigen named MGb1 to screen a phage-displayed random peptide library fused with coat protein pIII in order to get some information on mimotopes.

Methods: Through affinity enrichment and ELISA screening, positive clones of phages were amplified. 10 phage clones were selected after three rounds of biopanning and the ability of specific binding of the positive phage clones to MGb1-Ab were detected by ELISA assay (DNA sequencing was performed and the amino acid sequences were deduced) By blocking test, specificity of the mimic phage epitopes was identified.

Results: There were approximately 200 times of enrichment about the titer of bound phages after three rounds of biopanning procedures. DNA of 10 phage clones after the third biopanning was assayed and the result showed that the positive clones had a specific binding activity to MGb1-Ab and a weak ability of binding to control mAb or to mouse IgG. DNA sequencing of 10 phage clones was performed and the amino acid sequences were deduced. According to the homology of the amino acid sequences of the displayed peptides, most of the phage clones had motifs of H(x)Q or L(x)S. And these 10 phage clones could also partly inhibit the binding of MGb1-Ab to gastric cancer cell KATO-III. The percentage of blocking was from (21.0+/-1.6) % to (39.0+/-2.7) %.

Conclusion: Motifs of H(x)Q and L(x)S selected and identified show a high homology in the mimic epitopes of gastric cancer associated antigen. There may be one or more clones which can act as candidates of tumor vaccines.

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Figures

Figure 1
Figure 1
Binding of phages to MGb1-Ab by ELISA at the third round.
Figure 2
Figure 2
Electrophoresis of single strand DNA of phage clones (lanes1-10: Single strand DNA of phage clones; lane M: M13 single strand DNA marker).
Figure 3
Figure 3
Amino acid sequences of inserts in phage clones.
Figure 4
Figure 4
The blocking percentage of phage display peptides to MGb1-Ag by ELISA.
See this image and copyright information in PMC

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