Journal: bioRxiv

Article Title: Use of Epivolve phage display to generate a monoclonal antibody with opsonic activity directed against a subdominant epitope on extracellular loop 4 of Treponema pallidum BamA (TP0326)

doi: 10.1101/2023.05.13.540667

Figure Lengend Snippet: A peptide incorporating a non-native amino acid at a desired site is used to pan a scFv phage library for peptide binders. Phages that bind the modified, but not the native, peptide undergo AXM mutagenesis to generate phages that recognize the native peptide +/-the modified peptide with high affinity.

Article Snippet: The template plasmid for all phage display libraries is a derivative of the phagemid pIT2 ( ) with a human single-chain variable fragment Ab (scFv) fused to the coat protein III of bacteriophage M13, constructed at and kindly supplied by AxioMx Inc., an Abcam Company (“Abcam”).

Techniques: Modification, Mutagenesis

Journal: Biotechnology Progress

Article Title: High throughput automated microbial bioreactor system used for clone selection and rapid scale‐down process optimization

doi: 10.1002/btpr.2534

Figure Lengend Snippet: Process development and reproducibility tests in ambr 15f system. A : Growth curve during fedbatch fermentations of E. coli MC4100 cells grown in ambr 15f system. Cultures were induced with IPTG (1 mM) and fed with 40% (w/w) glycerol when carbon source was exhausted, n = 24 vessels Protein L HPLC results showing the scFv1 concentration in the periplasmic (P) fractions hours postinduction, n = 24 for 40 and 63 h; n = 6 for 32 and 48 h. B : Exponential growth phase of E. coli MC4100 cells from duplicate experiments 1 (▪) and 2 (○), n = 24 respectively.

Article Snippet: E. coli MC4100 cells expressing two single chain variable fragment antibodies (scFv). scFv raised against omega peptide of β‐galactosidase, designated as scFv 1 (courtesy of Cobra Biologics), scFv anti‐c‐Met, designated as scFv 2 (courtesy of James Austerberry, University of Manchester).

Techniques: Concentration Assay

Journal: Biotechnology Progress

Article Title: High throughput automated microbial bioreactor system used for clone selection and rapid scale‐down process optimization

doi: 10.1002/btpr.2534

Figure Lengend Snippet: Volume distribution of E. coli MC4100 cells expressing scFv 1 in fed‐batch fermentations grown in ambr 15f system. Samples from CS1 (•, ▪, filled triangles) and CS2 (○, □, △) A : pre exposure to high shear forces and B : post exposure to high shear forces.

Article Snippet: E. coli MC4100 cells expressing two single chain variable fragment antibodies (scFv). scFv raised against omega peptide of β‐galactosidase, designated as scFv 1 (courtesy of Cobra Biologics), scFv anti‐c‐Met, designated as scFv 2 (courtesy of James Austerberry, University of Manchester).

Techniques: Expressing, Shear

Journal: Biotechnology Progress

Article Title: High throughput automated microbial bioreactor system used for clone selection and rapid scale‐down process optimization

doi: 10.1002/btpr.2534

Figure Lengend Snippet: Scale‐down comparison of the ambr 15f and Multifors fermenters. A : Growth profile from fedbatch fermentations of E. coli MC4100 grown in parallel in ambr 15f (△, filled triangles) and Multifors bioreactors (□,▪), cascade control strategy A for filled symbols and B for the empty symbols. Number ofreplicates in Multifors = 2 and ambr 15f = 12. B : Protein L HPLC results showing the scFv 1 concentration inthe medium (M) and periplasmic (P) fractions from the ambr 15f (□) and Multifors (▪) fermentations at 18 h postinduction.

Article Snippet: E. coli MC4100 cells expressing two single chain variable fragment antibodies (scFv). scFv raised against omega peptide of β‐galactosidase, designated as scFv 1 (courtesy of Cobra Biologics), scFv anti‐c‐Met, designated as scFv 2 (courtesy of James Austerberry, University of Manchester).

Techniques: Comparison, Control, Concentration Assay

Journal: Biotechnology Progress

Article Title: High throughput automated microbial bioreactor system used for clone selection and rapid scale‐down process optimization

doi: 10.1002/btpr.2534

Figure Lengend Snippet: Comparison of feeding strategies and carbon source on E. coli MC4100 cells grown fed‐batch fermentation in ambr 15f system. A : Cells were fed with 40% (w/w) glycerol (▪,□) or 40% (w/w) glucose (○,•) using bolus additions (empty symbols) or exponential feeding (filled symbols) until they reach an OD of 100, then constant feeding (3.2 mL L ‐1 h ‐1 ) was used. n = 6 for each condition tested. B : scFv 1 concentration in the extracellular medium (M), periplasmic (P) and cytoplasm (C) fractions determined by HiTrap protein L HPLC. Vessels quantified/analyzed n = 6 for 16 and 40 h postinduction and n = 2 for 0 and 22 h postinduction.

Article Snippet: E. coli MC4100 cells expressing two single chain variable fragment antibodies (scFv). scFv raised against omega peptide of β‐galactosidase, designated as scFv 1 (courtesy of Cobra Biologics), scFv anti‐c‐Met, designated as scFv 2 (courtesy of James Austerberry, University of Manchester).

Techniques: Comparison, Concentration Assay

Journal: Biotechnology Progress

Article Title: High throughput automated microbial bioreactor system used for clone selection and rapid scale‐down process optimization

doi: 10.1002/btpr.2534

Figure Lengend Snippet: Screening of alternative strains and molecules in ambr 15f system. A : Growth curves during fedbatch fermentations of E. coli MC4100 cells expressing scFv 1 (○), scFv 2 (▪) and scFv 2 * (filled diamonds) ( n = 8). B : scFv 1 concentration in the periplasm (●), cytoplasm (▪) and extracellular medium (filled triangles) determined by protein L HPLC at 0, 15, 23, and 39 h post induction at harvest (vessels quantified/analyzed: n = 2 for 0 and 23 h and n = 8 for 15 and 39 h). C : Growth curves during fed‐batch fermentations of E. coli BL21 (DE3) cells expressing scFv 3 (S) (▪) scFv 3 (I) (□) and hGH (C) (●) hGH (I & P) (○), n = 6 for each molecule. D : Normalised protein concentration at harvest (42 h postinduction) determined by IMAC HPLC, vessels quantified/analyzed: n = 6 for each molecule.

Article Snippet: E. coli MC4100 cells expressing two single chain variable fragment antibodies (scFv). scFv raised against omega peptide of β‐galactosidase, designated as scFv 1 (courtesy of Cobra Biologics), scFv anti‐c‐Met, designated as scFv 2 (courtesy of James Austerberry, University of Manchester).

Techniques: Expressing, Concentration Assay, Protein Concentration

Journal: Biotechnology Progress

Article Title: High throughput automated microbial bioreactor system used for clone selection and rapid scale‐down process optimization

doi: 10.1002/btpr.2534

Figure Lengend Snippet: Fermentation Parameters from E. coli MC4100 Expressing a scFv 1 Grown in Minimal Medium with Glucose or Glycerol as Carbon Source

Article Snippet: E. coli MC4100 cells expressing two single chain variable fragment antibodies (scFv). scFv raised against omega peptide of β‐galactosidase, designated as scFv 1 (courtesy of Cobra Biologics), scFv anti‐c‐Met, designated as scFv 2 (courtesy of James Austerberry, University of Manchester).

Techniques: Expressing