Erratum to “Virtual mutation and directional evolution of anti-amoxicillin ScFv antibody for immunoassay of penicillins in milk” [Anal. Biochem. 517 (2017) 9–17]

4区 · 生物学

Article

作者: Dong, Jun ; Liu, Jing ; Qi, Yong Hua ; Zhao, Guo Xian ; Wang, Geng Nan ; He, Xin ; Duan, Chang Fei ; Wang, Jian Ping

Two panels from Fig. 3, C and F, were omitted.The corrected figure is provided.

2017-04-01·Nuclear Medicine and Biology4区 · 医学

PET imaging of 64Cu-DOTA-scFv-anti-PSMA lipid nanoparticles (LNPs): Enhanced tumor targeting over anti-PSMA scFv or untargeted LNPs

4区 · 医学

Article

作者: Li, Lin ; Crow, Desiree ; Wong, Patty ; Colcher, David ; Chea, Junie ; Bowles, Nicole ; Yazaki, Paul J ; Delgado, Melissa K ; Wong, Jeffrey Y C ; Channappa, Divya ; Shively, John E ; Poku, Erasmus ; Szpikowska, Barbara

INTRODUCTIONSingle chain (scFv) antibodies are ideal targeting ligands due to their modular structure, high antigen specificity and affinity. These monovalent ligands display rapid tumor targeting but have limitations due to their fast urinary clearance.METHODSAn anti-prostate membrane antigen (PSMA) scFv with a site-specific cysteine was expressed and evaluated in a prostate cancer xenograft model by Cu-64 PET imaging. To enhance tumor accumulation, the scFv-cys was conjugated to the co-polymer DSPE-PEG-maleimide that spontaneously assembled into a homogeneous multivalent lipid nanoparticle (LNP).RESULTSThe targeted LNP exhibited a 2-fold increase in tumor uptake compared to the scFv alone using two different thiol ester chemistries. The anti-PSMA scFv-LNP exhibited a 1.6 fold increase in tumor targeting over the untargeted LNP.CONCLUSIONSThe targeted anti-PSMA scFv-LNP showed enhanced tumor accumulation over the scFv alone or the untargeted DOTA-micelle providing evidence for the development of this system for drug delivery.ADVANCES IN KNOWLEDGE AND IMPLICATIONS FOR PATIENT CAREAnti-tumor scFv antibody fragments have not achieved their therapeutic potential due to their fast blood clearance. Conjugation to an LNP enables multivalency to the tumor antigen as well as increased molecular size for chemotherapy drug delivery.

2017-01-01·Analytical Biochemistry4区 · 生物学

Virtual mutation and directional evolution of anti-amoxicillin ScFv antibody for immunoassay of penicillins in milk

4区 · 生物学

Article

作者: Dong, Jun ; Liu, Jing ; Qi, Yong Hua ; Zhao, Guo Xian ; Wang, Geng Nan ; Duan, Chang Fei ; He, Xin ; Wang, Jian Ping

In this study, an anti-amoxicillin single chain variable fragment (ScFv) antibody was evolved by directional mutagenesis of a contact amino acid residue based on the analysis of virtual mutation. Comparison with its parental ScFv, the mutant showed highly improved affinity for 11 penicillins with up to 6-folds increased sensitivity. Then, its recognition mechanisms for the 11 drugs were studied by using molecular docking. Results showed that the mutant-penicillins intermolecular forces increased and the total binding energies decreased dramatically, which were responsible for the improvement of antibody sensitivity. The ScFv mutant was used to develop an indirect competitive enzyme linked immunosorbent assay for determination of the 11 drugs in milk. The limits of detection were in the range of 0.2-3.0 ng/mL, the crossreactivities were in the range of 31%-132%, and the recoveries from standards fortified blank milk were in the range of 65.7%-92.4%. This is the first study reporting the directional evolution of a ScFv antibody based on virtual mutation and the use of ScFv antibody for determination of penicillins in foods of animal origin.

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