Combine Phage Antibody Display Library Selection on Patient Tissue Specimens With Laser Capture Microdissection to Identify Novel Human Antibodies Targeting Clinically Relevant Tumor Antigens. Su Y, Bidlingmaier S, Lee NK, Liu B. Methods Mol Biol 1701:331-347, 2018. PMID: 29116514
Objective: to develop a technology that allows selection of phage antibody display libraries on tumor cells in situ residing in their natural tissue microenvironment.
Summary: Intracellular delivery of Immunotoxin was determined as follows: Immunotoxin was prepared by mixing biotinylated scFv with Streptavidin-ZAP (Cat. #IT-27) at a molar ratio of 1:1 and incubated on ice for 30 min. 50 μl of serially diluted immunotoxin was added to each well and incubated for 96 h at 37°C in 5% CO2. Cell growth medium were carefully removed from each well.
Dose: 100 μl of diluted CCK-8 was added to each well in the 96-well plates and incubated for 1–4 h at 37°C in 5% CO2. The absorbance was measured at 450 nm using a microtiter plate reader and the EC50 value determined using GraphPad Prism.
Characterization of the First Fully Human Anti-Tem1 SCFY in Models of Solid Tumor Imaging and Immunotoxin-Based Therapy.Yuan X, Yang M, Chen X, Zhang X, Sukhadia S, Musolino N, Bao H, Chen T, Xu C, Wang Q, Santoro S, Ricklin D, Hu J, Lin R, Yang W, Li Z, Qin W, Zhao A, Scholler N, Coukos G. Cancer Immunol Immunother 2018. PMID: 29313073
Objective: ScFv78 was conjugated with the ribosome-inactivating protein saporin (Streptavidin-ZAP) to evaluate whether scFv78 may be used as a vehicle for theTEM1-targeted delivery of toxins.
Summary: Site-specific, biotinylated scFv78 was conjugated with streptavidin-labeled saporin (Streptavidin-ZAP; Cat. #IT-27) by incubation at room temperature for 1h at a molar ratio of 4:1 (scFv78:ZAP).
Dose: Mouse endothelial cells (MS1) and MS1 cells transduced to express full-length human TEM1 (MS1-TEM1) were cultured in 96-well plates to 30% confluence and then incubated for 96h in the presence of 10-fold serially diluted Streptavidin-ZAP, scFv78, or scFv78-ZAP starting from 40nM down to 0.04nM. The data indicate that scFv78, the first fully human anti-TEM1 recombinant antibody, recognizes both human and mouse TEM1 and has unique and favorable features that are advantageous for the development of imaging probes or antibody-toxin conjugates for a large spectrum of human TEM1-positive solid tumors.
Substance P and Neurotensin in the Limbic System: Their Roles in Reinforcement and Memory Consolidation. Lénárd L, László K, Kertes E, Ollmann T, Péczely L, Kovács A, Kállai V, Zagorácz O, Gálosi R, Karádi Z. Neurosci Biobehav Rev 85:1-20, 2018. PMID: 28887225