TY - JOUR
T1 - Molecular analysis of cross-reactive human monoclonal antibody AE6F4 generated by in vitro immunization
T2 - Epitope mapping of AE6F4 antibody on 14-3-3 family proteins and cytokeratin 8
AU - Ichikawa, Akira
AU - Shim, Sun Yup
AU - Katakura, Yoshinori
AU - Teruya, Kiichiro
AU - Hashizume, Shuichi
AU - Shirahata, Sanetaka
PY - 2001
Y1 - 2001
N2 - We reported previously that adenocarcinoma-reactive human monoclonal antibody AE6F4, which had been generated by in vitro immunization method, recognizes both 14-3-3 protein and cytokeratin 8 (CK8). In this study, to analyze the cross-reactivity of AE6F4 antibody, epitopes of AE6F4 antibody on 14-3-3 proteins and CK8 were studied by using synthetic linear peptide scanning technology. To determine the locations of B cell epitope, 48 and 95 of decapeptides covering the entire 14-3-3 proteins and CK8, respectively, were synthesized and binding to AE6F4 antibody was examined by ELISA. The AE6F4 antibody was strongly reactive to peptides containing amino acid sequences TLWTSDTQGD in 14-3-3 proteins and INFLRQLYEE in CK8. These results indicate that AE6F4 antibody can recognize the different peptide sequences in 14-3-3 proteins and CK8.
AB - We reported previously that adenocarcinoma-reactive human monoclonal antibody AE6F4, which had been generated by in vitro immunization method, recognizes both 14-3-3 protein and cytokeratin 8 (CK8). In this study, to analyze the cross-reactivity of AE6F4 antibody, epitopes of AE6F4 antibody on 14-3-3 proteins and CK8 were studied by using synthetic linear peptide scanning technology. To determine the locations of B cell epitope, 48 and 95 of decapeptides covering the entire 14-3-3 proteins and CK8, respectively, were synthesized and binding to AE6F4 antibody was examined by ELISA. The AE6F4 antibody was strongly reactive to peptides containing amino acid sequences TLWTSDTQGD in 14-3-3 proteins and INFLRQLYEE in CK8. These results indicate that AE6F4 antibody can recognize the different peptide sequences in 14-3-3 proteins and CK8.
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U2 - 10.1023/A:1014060113186
DO - 10.1023/A:1014060113186
M3 - Article
C2 - 19003320
AN - SCOPUS:0035686102
VL - 36
SP - 101
EP - 107
JO - Cytotechnology
JF - Cytotechnology
SN - 0920-9069
IS - 1-3
ER -