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ADD reaction kinetics results paragraphs

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Nathan Dwarshuis 2021-07-27 17:42:11 -04:00
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tex/thesis.tex
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@ -96,6 +96,7 @@
\newacronym{tmb}{TMB}{3,3',5,5'-Tetramethylbenzidine}
\newacronym{gvhd}{GVHD}{graft-vs-host disease}
\newacronym{bcma}{BCMA}{B-cell maturation antigen}
\newacronym{di}{DI}{deionized}
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
% SI units for uber nerds
@ -951,6 +952,8 @@ transferred to another 96 well plate wherein expansion continued.
% METHOD snb decay curve generation and analysis (including the equation used to
% fit the data)
% METHOD add reaction kinetics diffusion mathy stuff
\subsection{Luminex Analysis}
Luminex was performed using a \product{ProcartaPlex kit}{\thermo}{custom} for
@ -1148,7 +1151,38 @@ higher, further showing that the decay of \gls{snb} is not a concern.
\label{fig:dms_flowchart}
\end{figure*}
% RESULT add paragraph explaining the reaction kinetics stuff
We also investigated the reaction kinetics of all three coating steps.
To quantify the reaction kinetics of the biotin binding step, we reacted
multiple batches of microcarriers at \gls{rt} with
\gls{20}{\mg\of{\carrier}\per\ml} with \gls{snb} in parallel and sacrificially
analyzed each at varying timepoints using the \gls{haba} assay. This was
performed at two different concentrations. We observed that for either
concentration, the reaction was over in \SIrange{20}{30}{\minute}
(\cref{fig:dms_biotin_rxn_mass}). Furthermore, when put in terms of fraction of
input \gls{snb}, we observed that the curves are almost identical
(\cref{fig:dms_biotin_rxn_frac}). Given this, the reaction step for biotin
attached was set to \gls{30}{\minute}.
% TODO these numbers might be totally incorrect
Next, we quantified the amount of \gls{stp} reacted with the surface of the
biotin-coated microcarriers. Different batches of biotin-coated \glspl{dms} were
coated with \SI{40}{\ug\per\ml} \gls{stp} and sampled at various timepoints
using the \gls{bca} assay to indirectly quantify the amount of attached
\gls{stp} mass. We found this reaction took \SI{45}{\minute}
(\cref{fig:dms_stp_per_time}).
% TODO find real numbers for this
Finally, we used the reaction data from the \gls{stp} binding curve to estimate
the \gls{mab} binding curve. Assuming a quasi-steady-state paradigm, we
estimated that the diffusion rate coefficient for the microcarriers was
{\#}{diffusion rate}. Using this diffusion rate and the maximum mass of
\glspl{mab} bound the microcarriers (\cref{fig:mab_coating}), we estimated that
the \gls{mab} reaction should proceed in {\#}{mab curve}.
% TODO add additional paragraph about how this diffusion coefficient was used to
% estimate the wash step times.
\begin{figure*}[ht!]
\begingroup