Minimal motif mapping of a known epitope on human zona pellucida protein-4 using a peptide biosynthesis strategy.
An important step in the development of a human zona pellucida (huZP) peptide vaccine is to define the minimal amino acid motif for a mapped B cell epitope peptide within huZP4. Identification of this minimal motif is necessary to remove an overlapping T cell epitope that induces a pathogenic T cell response. Here we describe motif (PLTLEL(314-319)) mapping of an 18mer B cell epitope peptide(308-325) on huZP4 protein (previously known as huZP1/ZPB protein), achieved using a set of 22 biosynthetic 8mer peptides fused with truncated glutathione S-transferase (GST) or truncated streptavidin protein, and detected using rabbit anti-porcine zona pellucida (pZP) IgG. The immunogenicity of the B cell epitope peptide was evaluated in rabbits using expressed B cell epitope peptide fused with truncated streptavidin as the antigen. This construct elicited high titer antibody to the 18mer B cell epitope peptide, with reactivity to native human ZP, the biosynthetic 18mer peptide and the 18mer peptide GST fusion protein.
Xu
Wan-Xiang
WX
Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China. xuwx@sippr.stc.sh.cn
He
Ya-Ping
YP
Tang
Hai-Ping
HP
Jia
Xiao-Feng
XF
Ji
Chao-Neng
CN
Gu
Shao-Hua
SH
Sun
Xiao-Xi
XX
Xie
Yi
Y
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D004527:Egg Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D005260:Female; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology*; D011247:Pregnancy; D015202:Protein Engineering; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D019809:Streptavidin / Q000235:genetics / Q000378:metabolism; D013552:Swine; D000072216:Zona Pellucida Glycoproteins
D004527:Egg Proteins; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C082833:ZP4 protein, human; D000072216:Zona Pellucida Glycoproteins; D019809:Streptavidin; D005982:Glutathione Transferase