Effect of linker length between variable domains of single chain variable fragment antibody against daidzin on its reactivity.
The peptide linker between variable domains of heavy (VH) and light (VL) chains is one of important factors that influence the characteristics of scFv, including binding activity and specificity against target antigen. The scFvs against daidzin (DZ-scFvs) with different linker lengths were constructed in the format of VH-(GGGGS)n-VL (n = 1, 3, 5, and 7). They were expressed in the hemolymph of silkworm larvae using the Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid DNA system, and their reactivity against daidzin and related compounds were evaluated using an indirect competitive enzyme-linked immunosorbent assay (icELISA), which is applicable for quantitative analysis of daidzin. The results showed that the reactivity of scFvs against daidzin was increased, whereas specificity slightly decreased when their peptide linker was lengthened. These results suggested that the linker length of DZ-scFvs contributes to its reactivity. In addition, the results emphasize that the linker length could control the reactivity of DZ-scFvs.
a Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences , Kyushu University , Fukuoka , Japan.
Gorawit
G
Yusakul
a Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences , Kyushu University , Fukuoka , Japan.
Seiichi
S
Sakamoto
b Faculty of Pharmacy, Department of Pharmaceutical Botany , Mahidol University , Bangkok , Thailand.
Benyakan
B
Pongkitwitoon
a Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences , Kyushu University , Fukuoka , Japan.
Hiroyuki
H
Tanaka
a Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences , Kyushu University , Fukuoka , Japan.
Satoshi
S
Morimoto
D020816:Amino Acid Motifs; D000818:Animals; D000918:Antibody Specificity; D012831:Bombyx / Q000737:chemistry / Q000276:immunology; D004247:DNA / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D006458:Hemolymph / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D007529:Isoflavones / Q000737:chemistry* / Q000276:immunology; D007814:Larva / Q000737:chemistry / Q000276:immunology*; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D013329:Structure-Activity Relationship
D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007529:Isoflavones; D011994:Recombinant Proteins; C013908:daidzin; D004247:DNA
BmNPV bacmid DNA system; daidzin; icELISA; linker length; scFv