Journal: bioRxiv

Article Title: Multiparametric optimization of human primary B-cell cultures using Design of Experiments

doi: 10.1101/2025.01.16.633474

Figure Lengend Snippet: Stepwise gating of human PBMCs, exemplifying the lymphocyte/singlet-FSC/singlet-SSC/live/CD19 + hierarchy used in subsequent analyses.

Article Snippet: Human CD19 + B cells were purified from buffy coats using the StraightFrom Buffy Coat CD19 MicroBead purification Kit (Miltenyi Biotec, cat. #130-114-974) following the manufactures instructions.

Techniques:

Journal: bioRxiv

Article Title: Multiparametric optimization of human primary B-cell cultures using Design of Experiments

doi: 10.1101/2025.01.16.633474

Figure Lengend Snippet: ( A-D ) Pareto plot of parameter effect sizes on IgE ( A ), IgA ( B ), IgG1 ( C ), and IgG3 ( D ) detection in the medium of DOE 6 cultures measured by TRIFMA. ( E-G ) Measured IgG1 and IgG3 in culture medium of each sample from DOE 6 where color and mean indicated by dotted lines are specific for time ( E ), level of IL-4 ( F ), N.40 expression ( G ). ( H ) As ( E ) but for measuring IgG1 and IgA. ( I ) Flow plotting IgD vs. CD27 of CD19 + live singlet lymphocytes after 10 days of naïve B-cell iGB culturing to illustrate the difference of their activation and differentiation depending on the stimuli provided, with the feeder-cell expression level of CD40L being a very apparent contributor. ( J ) as ( I ) but plotting CD38 vs. CD95.

Article Snippet: Human CD19 + B cells were purified from buffy coats using the StraightFrom Buffy Coat CD19 MicroBead purification Kit (Miltenyi Biotec, cat. #130-114-974) following the manufactures instructions.

Techniques: Expressing, Activation Assay