REF add some notes to myself
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@ -698,6 +698,7 @@ Therefore, the proposed microcarrier system should enable greater expansion and
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better retention of memory phenotype compared to current bead-based methods.
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better retention of memory phenotype compared to current bead-based methods.
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\section{methods}
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\section{methods}
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\subsection{dms functionalization}
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\subsection{dms functionalization}
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\begin{figure*}[ht!]
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\begin{figure*}[ht!]
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@ -834,6 +835,8 @@ Cells on the \glspl{dms} were visualized by adding \SI{0.5}{\ul}
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\product{\acd{45}-\gls{af647}}{\bl}{368538}, incubating for \SI{1}{\hour}, and
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\product{\acd{45}-\gls{af647}}{\bl}{368538}, incubating for \SI{1}{\hour}, and
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imaging on a spinning disk confocal microscope.
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imaging on a spinning disk confocal microscope.
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% METHOD mtt staining and interior quantification
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\subsection{quantification of apoptosis using Annexin-V}
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\subsection{quantification of apoptosis using Annexin-V}
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Apoptosis was quantified using \gls{anv} according to the manufacturer's
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Apoptosis was quantified using \gls{anv} according to the manufacturer's
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@ -905,6 +908,7 @@ Briefly, cells were washed once and stained with \product{biotinylated
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BD Accuri. Readout was percent \gls{pe}+ cells as compared to secondary controls
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BD Accuri. Readout was percent \gls{pe}+ cells as compared to secondary controls
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(\gls{pe}-\gls{stp} with no \gls{ptnl}).
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(\gls{pe}-\gls{stp} with no \gls{ptnl}).
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% TODO add BCMA-CAR stuff
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\subsection{car plasmid and lentiviral transduction}
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\subsection{car plasmid and lentiviral transduction}
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The anti-CD19-CD8-CD137-CD3z \gls{car} with the EF1$\upalpha$
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The anti-CD19-CD8-CD137-CD3z \gls{car} with the EF1$\upalpha$
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@ -924,8 +928,6 @@ plates and incubated for another \SI{24}{\hour}. Cells and beads/\glspl{dms}
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were removed from the retronectin plates using vigorous pipetting and
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were removed from the retronectin plates using vigorous pipetting and
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transferred to another 96 well plate wherein expansion continued.
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transferred to another 96 well plate wherein expansion continued.
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\subsection{statistical analysis}
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\subsection{statistical analysis}
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For 1-way \gls{anova} analysis with Tukey multiple comparisons test,
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For 1-way \gls{anova} analysis with Tukey multiple comparisons test,
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@ -947,7 +949,8 @@ lack-of-fit tests where replicates were present (to assess model fit in the
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context of pure error). Statistical significance was evaluated at $\upalpha$ =
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context of pure error). Statistical significance was evaluated at $\upalpha$ =
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0.05.
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0.05.
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% TODO add meta-analysis section
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% METHOD add meta-analysis
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% METHOD add Grex
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\section{results}
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\section{results}
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@ -976,9 +979,8 @@ noted that the maximal \gls{mab} binding capacity occurred near \SI{50}{\nmol}
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biotin input (which corresponded to \SI{2.5}{\nmol\per\mg\of{\dms}}) thus we
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biotin input (which corresponded to \SI{2.5}{\nmol\per\mg\of{\dms}}) thus we
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used this in subsequent processes.
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used this in subsequent processes.
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% TODO add paragraph explaining the qc stuff
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% RESULT add paragraph explaining the qc stuff
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% RESULT add paragraph explaining the reaction kinetics stuff
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% TODO add paragraph explaining the reaction kinetics stuff
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% TODO flip the rows of this figure (right now the text is backward)
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% TODO flip the rows of this figure (right now the text is backward)
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\begin{figure*}[ht!]
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\begin{figure*}[ht!]
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@ -1135,7 +1137,6 @@ cells after \SI{14}{\day} were on the interior surface of the \glspl{dms}
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\label{fig:dms_flowchart}
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\label{fig:dms_flowchart}
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\end{figure*}
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\end{figure*}
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% TODO add this to the methods section
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% TODO double check the timing of this experiment (it might not be day 14)
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% TODO double check the timing of this experiment (it might not be day 14)
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% TODO provide the regression results and coefficients from this
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% TODO provide the regression results and coefficients from this
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\begin{figure*}[ht!]
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\begin{figure*}[ht!]
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@ -1290,9 +1291,17 @@ showing that migration was likely independent of \gls{car} transduction.
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\subcap{fig:car_production_migration}{Endpoint plot for transmigration assay
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\subcap{fig:car_production_migration}{Endpoint plot for transmigration assay
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with \gls{anova}.} All data is from T cells expanded for \SI{14}{\day}.
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with \gls{anova}.} All data is from T cells expanded for \SI{14}{\day}.
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}
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}
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\label{fig:dms_phenotype}
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\label{fig:car_production}
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\end{figure*}
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\end{figure*}
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% FIGURE bcma results
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\subsection{DMSs efficiently expand T cells in Grex bioreactors}
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% FIGURE grex
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% FIGURE luminex
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% FIGURE grex + car
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\subsection{DMSs do not leave antibodies attached to cell product}
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\subsection{DMSs do not leave antibodies attached to cell product}
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We asked if \glspl{mab} from the \glspl{dms} detached from the \gls{dms} surface
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We asked if \glspl{mab} from the \glspl{dms} detached from the \gls{dms} surface
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@ -1625,6 +1634,8 @@ provide these benefits.
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\subsection{DOE shows optimal conditions for expanded potent T cells}
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\subsection{DOE shows optimal conditions for expanded potent T cells}
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% TABLE luminex panel used
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% TODO this plots proportions look dumb
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% TODO this plots proportions look dumb
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\begin{figure*}[ht!]
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\begin{figure*}[ht!]
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\begingroup
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\begingroup
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@ -1699,7 +1710,7 @@ provide these benefits.
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\label{fig:doe_responses}
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\label{fig:doe_responses}
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\end{figure*}
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\end{figure*}
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% TODO add DOE regression tables
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% TABLE DOE regression results
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% TODO this section header sucks
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% TODO this section header sucks
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\subsection{AI modeling reveals highly predictive species}
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\subsection{AI modeling reveals highly predictive species}
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