TY - JOUR
T1 - The effect of varying the peptide linker length in a single chain variable fragment antibody against wogonin glucuronide
AU - Paudel, Madan Kumar
AU - Sakamoto, Seiichi
AU - Van Huy, Le
AU - Tanaka, Hiroyuki
AU - Miyamoto, Tomofumi
AU - Morimoto, Satoshi
PY - 2017/6/10
Y1 - 2017/6/10
N2 - Peptide linkers of three different lengths were constructed to join the variable regions of the heavy chain (VH) and the light chain (VL) in a single-chain variable fragment antibody (scFv) specific for wogonin glucuronide (Wgn) that has the structure VH-(GGGGS)n–VL (n = 3, 5, or 7). The scFv antibodies, which were expressed in Escherichia coli, were derived from an anti-Wgn monoclonal antibody (315A). An indirect competitive enzyme-linked immunosorbent assay (icELISA) was used to evaluate their reactivity and sensitivity, which is also used for quantitative analysis of Wgn. Our results, showed that the reactivity and specificity of the three different scFvs were, in fact, similar. Subsequently, the scFv having a VH-(GGGGS)3–VL linker which was slightly better that other two scFvs against Wgn, was applied to indirect competitive ELISA (icELISA) to analyze Scutellariae Radix (S. Radix). The utility of the icELISA was demonstrated for quality control and analysis of S. Radix in this report.
AB - Peptide linkers of three different lengths were constructed to join the variable regions of the heavy chain (VH) and the light chain (VL) in a single-chain variable fragment antibody (scFv) specific for wogonin glucuronide (Wgn) that has the structure VH-(GGGGS)n–VL (n = 3, 5, or 7). The scFv antibodies, which were expressed in Escherichia coli, were derived from an anti-Wgn monoclonal antibody (315A). An indirect competitive enzyme-linked immunosorbent assay (icELISA) was used to evaluate their reactivity and sensitivity, which is also used for quantitative analysis of Wgn. Our results, showed that the reactivity and specificity of the three different scFvs were, in fact, similar. Subsequently, the scFv having a VH-(GGGGS)3–VL linker which was slightly better that other two scFvs against Wgn, was applied to indirect competitive ELISA (icELISA) to analyze Scutellariae Radix (S. Radix). The utility of the icELISA was demonstrated for quality control and analysis of S. Radix in this report.
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U2 - 10.1016/j.jbiotec.2017.04.002
DO - 10.1016/j.jbiotec.2017.04.002
M3 - Article
C2 - 28392422
AN - SCOPUS:85017589560
SN - 0168-1656
VL - 251
SP - 47
EP - 52
JO - Journal of Biotechnology
JF - Journal of Biotechnology
ER -