Journal: Biotechnology Progress

Article Title: Optimization of multi‐column chromatography for capture and polishing at high protein load

doi: 10.1002/btpr.70047

Figure Lengend Snippet: Optimization plots of the three different Protein A resins studied for the capture step of mAb. (a), (b), and (c) are the Pareto fronts for MSS, MSPrismA, and MSSpcc, respectively. The optimization was done for batch mode (at c mAb of 5 g/L) and continuous mode (at c mAb of 2.5, 5, 7.5, and 10 g/L). Numbers 1, 3, 4, and 5 represent the results for continuous mode at c mAb of 2.5, 5, 7.5, and 10 g/L, respectively; number 2 represents the results for batch mode at c mAb of 5 g/L. (d) Comparison of the optimizations for continuous chromatography of the three different resins at a 5 g/L concentration of mAb. Concentrations of 7.5 and 10 g/L represent an extrapolation of the feed concentrations for model calibration.

Article Snippet: 1 mL HiTrap® columns of ProA resins Mab Select SuRe (MSS), Mab Select PrismA (MSPrismA), and Mab Select SuRe pcc (MSSpcc) and CEX resins CaptoTM S ImpAct (CaptoS Imp) and SP Sepharose Fast Flow (SP Seph FF) were purchased from Cytiva, Uppsala, Sweden.

Techniques: Comparison, Chromatography, Concentration Assay

Journal: Biotechnology Progress

Article Title: Optimization of multi‐column chromatography for capture and polishing at high protein load

doi: 10.1002/btpr.70047

Figure Lengend Snippet: Experimental validation of the continuous chromatography model for the capture step with a pure sample of mAb using MSPrismA. The initial concentration is 5 g/L and the loading flow rate is 0.71 mL/min. (a) Total chromatogram; (b) Zoom in on the steady‐state of the cyclic operation of the 3C‐PCC; (c) Model data for the same cyclic period as shown in B. In (a) and (b), black and red represent the concentration observed in UV1 and UV2, respectively. In (c), black, red, and blue represent the outlet concentrations predicted from the model for columns 1, 2, and 3, respectively.

Article Snippet: 1 mL HiTrap® columns of ProA resins Mab Select SuRe (MSS), Mab Select PrismA (MSPrismA), and Mab Select SuRe pcc (MSSpcc) and CEX resins CaptoTM S ImpAct (CaptoS Imp) and SP Sepharose Fast Flow (SP Seph FF) were purchased from Cytiva, Uppsala, Sweden.

Techniques: Biomarker Discovery, Chromatography, Concentration Assay

Journal: Biochemical Engineering Journal

Article Title: Economic optimization of antibody capture through Protein A affinity nanofiber chromatography

doi: 10.1016/j.bej.2023.109141

Figure Lengend Snippet: Fig. 1. Dynamic binding capacity (DBC) at 10% breakthrough. A) Comparison between MabSelect PrismA (dashed line) and Fibro PrismA (solid line) with log residence time. The residence time for Fibro™ PrismA was < 1 min and for MabSelect™ PrismA 4–5 min. B) Fibro PrismA DBC against residence time fitted by a two- parameter power function (dashed line). IgG1(•); IgG4–1 (∎); IgG4–2 (▴); IgG4–3(▾) and trivalent Fc (◆).

Article Snippet: The resin column, HiScreen MabSelectTM PrismA (4.7 mL) with a 15–40 nm pore diameter, 10 cm bed height and 40 m2 g-1 surface area, was also purchased from Cytiva (Sweden).

Techniques: Binding Assay, Comparison

Journal: Biochemical Engineering Journal

Article Title: Economic optimization of antibody capture through Protein A affinity nanofiber chromatography

doi: 10.1016/j.bej.2023.109141

Figure Lengend Snippet: Fig. 2. Binding signal between different concentrations of proteins and PrismA ligands. A) Illustration of the SPR technique to determine static binding capacity. B) Experimental data and the fit to a Langmuir isotherm for binding signal against protein concentration. C) Error margins of parameter Qmax for each mAb. D) Error margins of parameter keq for each mAb.

Article Snippet: The resin column, HiScreen MabSelectTM PrismA (4.7 mL) with a 15–40 nm pore diameter, 10 cm bed height and 40 m2 g-1 surface area, was also purchased from Cytiva (Sweden).

Techniques: Binding Assay, Protein Concentration

Journal: Biochemical Engineering Journal

Article Title: Economic optimization of antibody capture through Protein A affinity nanofiber chromatography

doi: 10.1016/j.bej.2023.109141

Figure Lengend Snippet: Fig. 4. Protein impurity clearance by MabSelect PrismA at 0.25 CV min-1 (∎) and Fibro PrismA at 40 MV min-1(•) as a function of the load density. Aggregates reduction is shown on the left hand side for IgG1, IgG4–3 and Trivalent Fc; while host cell protein (HCP) reduction is on the right hand side for the same proteins.

Article Snippet: The resin column, HiScreen MabSelectTM PrismA (4.7 mL) with a 15–40 nm pore diameter, 10 cm bed height and 40 m2 g-1 surface area, was also purchased from Cytiva (Sweden).

Techniques:

Journal: Biochemical Engineering Journal

Article Title: Economic optimization of antibody capture through Protein A affinity nanofiber chromatography

doi: 10.1016/j.bej.2023.109141

Figure Lengend Snippet: Fig. 5. Comparison of chromatograms for different elution volumes (10 MV and 15 MV) with HiTrap (0.4 mL) and HiScreen (3.75 mL) Fibro™ PrismA devices using IgG4–3 as the model. A) Whole chromatograms starting from sample loading; B) Elution profile. HiTrap Fibro PrismA and HiScreen Fibro PrismA were operated using ¨Akta Pure 25 and ¨Akta Pure 150, respectively.

Article Snippet: The resin column, HiScreen MabSelectTM PrismA (4.7 mL) with a 15–40 nm pore diameter, 10 cm bed height and 40 m2 g-1 surface area, was also purchased from Cytiva (Sweden).

Techniques: Comparison

Journal: Biochemical Engineering Journal

Article Title: Economic optimization of antibody capture through Protein A affinity nanofiber chromatography

doi: 10.1016/j.bej.2023.109141

Figure Lengend Snippet: Fig. 6. Overlay of 200 cycles of IgG4–3 capture on Fibro PrismA for: A) Chromatograms; B) Delta pressure drop; C) Eluate peak width at half height; D) Peak height; E) Eluate volume; and F) Protein recovery.

Article Snippet: The resin column, HiScreen MabSelectTM PrismA (4.7 mL) with a 15–40 nm pore diameter, 10 cm bed height and 40 m2 g-1 surface area, was also purchased from Cytiva (Sweden).

Techniques:

Journal: Biochemical Engineering Journal

Article Title: Economic optimization of antibody capture through Protein A affinity nanofiber chromatography

doi: 10.1016/j.bej.2023.109141

Figure Lengend Snippet: Fig. 7. Eluate quality over 200 cycles of IgG4–3 capture on Fibro PrismA. A) Aggregate reduction; B) Log HCP reduction; C) Intensity Average Particle size; and D) Protein A ligand leakage level.

Article Snippet: The resin column, HiScreen MabSelectTM PrismA (4.7 mL) with a 15–40 nm pore diameter, 10 cm bed height and 40 m2 g-1 surface area, was also purchased from Cytiva (Sweden).

Techniques:

Journal: Biochemical Engineering Journal

Article Title: Economic optimization of antibody capture through Protein A affinity nanofiber chromatography

doi: 10.1016/j.bej.2023.109141

Figure Lengend Snippet: Fig. 9. Cost comparison between MabSelect PrismA and Fibro PrismA at manufacturing scale (2000 L). A) Total cost comparison for a single batch. Cost per gram mAb processed for up to 20 harvests at a manufacturing scale for B) A new membrane device is replaced every harvest; and C) The membrane device is replaced every 4 harvests (200 cycles).

Article Snippet: The resin column, HiScreen MabSelectTM PrismA (4.7 mL) with a 15–40 nm pore diameter, 10 cm bed height and 40 m2 g-1 surface area, was also purchased from Cytiva (Sweden).

Techniques: Comparison, Membrane

Journal: Biochemical Engineering Journal

Article Title: Economic optimization of antibody capture through Protein A affinity nanofiber chromatography

doi: 10.1016/j.bej.2023.109141

Figure Lengend Snippet: Fig. 8. Helium ion microscopy analysis of Fibro PrismA before and after 200 cycles. A), B) Unused Fibro PrismA. C), D) Outlet region of the Fibro PrismA after 200 cycles at two different resolutions. The red circles showed polymer beads of fibers resulting from the electrospinning process.

Article Snippet: The resin column, HiScreen MabSelectTM PrismA (4.7 mL) with a 15–40 nm pore diameter, 10 cm bed height and 40 m2 g-1 surface area, was also purchased from Cytiva (Sweden).

Techniques: Microscopy, Polymer

Journal: Biochemical Engineering Journal

Article Title: Economic optimization of antibody capture through Protein A affinity nanofiber chromatography

doi: 10.1016/j.bej.2023.109141

Figure Lengend Snippet: Fig. 10. Cost comparison between MabSelect PrismA and Fibro PrismA at A) Pilot scale (200 L) and B) Clinical Scale (500 L).

Article Snippet: The resin column, HiScreen MabSelectTM PrismA (4.7 mL) with a 15–40 nm pore diameter, 10 cm bed height and 40 m2 g-1 surface area, was also purchased from Cytiva (Sweden).

Techniques: Comparison

Journal: bioRxiv

Article Title: Enhancement of antibody thermostability and affinity by computational design in the absence of antigen

doi: 10.1101/2023.12.19.572421

Figure Lengend Snippet: (A) In 96-well format, DNA was normalized to match concentrations prior to transfection. (B) Expi293 cells (3 mL) were dispensed in 24-deep-well plates and transfected with DNA from the 96-well plate in A1, A2, B1, B2 quadrant format. (C) Seven days after transfection, cells were centrifuged and supernatant transferred to new 24-deep-well plates. An aliquot of supernatant (60 µL) was removed to measure antibody titer by Octet BLI Protein A prior to the addition of 200 µL 25% slurry (i.e., 50 µL settled) of MagSepharose PrismA Protein A beads. (D) Supernatant and Protein A beads were incubated with shaking at room temperature for 4 hours to immobilize antibodies on beads. (E) Using a 24-well plate magnet, supernatant was removed, Protein A beads with bound antibodies were washed, and antibodies were eluted. (F) Antibodies were then transferred back into 96-well format to the appropriate A1, A2, B1, B2 quadrants from which their DNA originated. Figure created with BioRender.

Article Snippet: A 25% slurry of MagSepharose PrismA Protein A magnetic beads (Cytiva #17550001) in 0.1% Tween-20/PBS was added at 200μL/well of supernatant, then incubated on a shaker at room temperature for 4 hours.

Techniques: Transfection, Incubation