Construction of multiform scFv antibodies using linker peptide.
Multiform single chain variable fragments (scFvs) including different length linker scFvs and bispecific scFv were constructed. The linker lengths of 0, 3, 5, 8, 12, and 15 amino acids between V(H) and V(L) of antideoxynivalenol (anti-DON) scFv were used to analyze the affinities of scFvs. The affinity constants of these scFvs increased when the linker was lower than 12 amino acids. The affinity constant would not change when the linker was longer than 12 amino acids. Fusion gene of anti-DON scFv and antizearalenone (anti-ZEN) scFv was also constructed through connection by a short peptide linker DNA to express a bispecific scFv. The affinity constants assay showed that the two scFvs of fusion bispecific scFv remained their own affinity compared to their parental scFvs. Competitive direct enzyme linked immunosorbent assay was used to detect DON and ZEN in contaminated wheat (Triticum aestivum L.) samples, and the results indicated that this bispecific scFv was applicable in DON and ZEN detection. This work confirmed that bispecific scFv could be successfully obtained, and might also have an application in diagnosing fungal infection, and breeding transgenic plants.
Wang
Shihua
S
Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Zheng
Cengjie
C
Liu
Ying
Y
Zheng
Huirong
H
Wang
Zonghua
Z
D000906:Antibodies / Q000032:analysis / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D018033:Antibodies, Bispecific / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000915:Antibody Affinity; D001483:Base Sequence; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005506:Food Contamination / Q000032:analysis; D015870:Gene Expression; D007135:Immunoglobulin Variable Region / Q000032:analysis / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D010455:Peptides / Q000737:chemistry*; D014255:Trichothecenes / Q000276:immunology / Q000633:toxicity; D014908:Triticum; D015025:Zearalenone / Q000276:immunology / Q000633:toxicity
D000906:Antibodies; D018033:Antibodies, Bispecific; D007135:Immunoglobulin Variable Region; D010455:Peptides; D014255:Trichothecenes; D015025:Zearalenone; C007262:deoxynivalenol