Author: Rogers, J.; Schoepp, R.J.; Schröder, O.; Clements, T.L.; Holland, T.F.; Li, J.Q.; Li, J.; Lewis, L.M.; Dirmeier, R.P.; Frey, G.J.; Tan, X.; Wong, K.; Woodnutt, G.; Keller, M.; Reed, D.S.; Kimmel, B.E.; Tozer, E.C.
Title: Rapid discovery and optimization of therapeutic antibodies against emerging infectious diseases Document date: 2008_5_13
ID: xkx56h0o_42
Snippet: Screening of these libraries was accomplished using a novel high throughput DNA display methodology (Fig. 1 ). This technique involves the intracellular expression of a Fab fragment as a fusion protein with a zinc finger. Because the zinc finger binds to its corresponding DNA binding domain on the plasmid, the fusion protein binds to the DNA encoding its genotype. Following cell lysis, this complex is panned against the antigen of interest. The f.....
Document: Screening of these libraries was accomplished using a novel high throughput DNA display methodology (Fig. 1 ). This technique involves the intracellular expression of a Fab fragment as a fusion protein with a zinc finger. Because the zinc finger binds to its corresponding DNA binding domain on the plasmid, the fusion protein binds to the DNA encoding its genotype. Following cell lysis, this complex is panned against the antigen of interest. The fusion protein -plasmid complex was found to be extremely stable by Biacore analysis (data not shown) with a half-life of $9 h at room temperature. Also of note, tests were done to ensure that no plasmid switching occurred at this time (i.e. the complex dissociating and the fusion protein binding to the wrong plasmid). Multiple separate screens were performed using Dynal beads coated with the spike protein, whole viral isolates, and infected cell lysates as antigens.
Search related documents:
Co phrase search for related documents- antigen cell lysate and infected antigen cell lysate: 1, 2
- bind domain and dna bind: 1, 2, 3, 4
- bind domain and fusion protein: 1, 2, 3, 4, 5, 6, 7, 8
- bind domain and high throughput: 1, 2, 3, 4
- bind domain and room temperature: 1, 2
- bind domain and spike protein: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60
- bind domain and zinc finger: 1
- cell lysate and complex dissociate: 1
- cell lysate and fusion protein: 1, 2, 3, 4, 5, 6, 7
- cell lysate and infected antigen cell lysate: 1, 2
- cell lysate and room temperature: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
- cell lysate and spike protein: 1, 2, 3
- cell lysis and fusion protein: 1, 2, 3
- cell lysis and high throughput: 1, 2
Co phrase search for related documents, hyperlinks ordered by date