Generation and analysis of the improved human HAL9/10 antibody phage display libraries.
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Cites methods from "Generation and analysis of the impr..."
...Two scFv naive universal libraries (HAL10 and HAL9 HAL10) were employed (32)....
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...45 10(9), respectively, were used for panning (32)....
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248 citations
Cites background from "Generation and analysis of the impr..."
...The theoretical size of universal libraries is now greater than 10(10) independent clones.(38,47,59,61,68,69) A particular type of antibody library is generated during guided phage display selection of human VH and VL repertoires using the counter chain of a murine monoclonal antibody as template....
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...Examples of na€ıve universal libraries are the human Fab library constructed by de Haard and colleagues at Dyax (now Shire),(40) the scFv libraries from Cambridge Antibody Technology (CAT),(39,61) scFv and Fab libraries from XOMA(59) or the HAL scFv libraries.(37,47)...
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References
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"Generation and analysis of the impr..." refers methods in this paper
...A volume of 180 μL phosphate-buffered 2 × YT-GA in a PP-MTP well was inoculated with 10 μL of the overnight culture and grown at 37°C and 800 rpm for 2 h. Bacteria were harvested by centrifugation for 10 min at 3,220 × g and 180 μL supernatants were removed....
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...Protean II Minigel system (BioRad Inc, München, Germany) according to [37]....
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...After transformation of XL1-Blue MRF’, overnight incubation on 25 cm 2 × YT-GAT plates and resuspension in 25 mL 2 × YT medium, glycerol stocks of each library were made using 800 μL bacteria solution (~1010 bacteria) and 200 μL glycerol (Roth, Karlsruhe) and stored at −80°C....
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...0,5 mL, Greiner, Frickenhausen, Germany) containing 150 μL 2 × YT-GA [37] were inoculated with the bacteria bearing scFv expressing phagemids....
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...Electroporation was performed with 0.1 cm prechilled cuvettes using a MicroPulser electroporator (BioRad, München) at 1.7 kV. Immediately, 1 mL prewarmed 37°C SOC medium pH 7.0 (2% (w/v) tryptone, 0.5% (w/v) yeast extract, 0.05% (w/v) NaCl, 20 mM Mg solution, 20 mM glucose) was added and incubated for 1 h at 600 rpm and 37°C. Transformation was plated out on 25 cm square Petri dishes with 2xYT-GAT agar (1,6% (w/v) Tryptone, 1% (w/v) yeast extract, 0,5% (w/v) NaCl, 100 mM glucose, 100 μg/mL ampiciline, 20 μg/mL tetracycline, 1.5% (w/v) agar-agar) and incubated overnight at 37°C....
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1,973 citations
"Generation and analysis of the impr..." refers methods in this paper
...antibody gene libraries are used: immune libraries and universal or “single-pot” libraries [24,25]....
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1,170 citations