diff --git a/code/microcarrier_diffusion_washing.m b/code/microcarrier_diffusion_washing.m index a468378..cd59d01 100644 --- a/code/microcarrier_diffusion_washing.m +++ b/code/microcarrier_diffusion_washing.m @@ -124,11 +124,11 @@ diff = 1000000; % init to some huge number while diff > tolerance t = linspace(t_0, t_f, 50); - Y = pdepe(m,@pde,@init,@bound,r,t); - C = Y(:,:,1); + Y = pdepe(m, @pde, @init, @bound, r, t); + C = Y(:, :, 1); % final concentration in center of carrier at final time - C_f_center = C(end,1); + C_f_center = C(end, 1); % test to see how far off the center is from bulk diff = C_f_center / C_Lbf - 1; @@ -166,13 +166,13 @@ function u0 = init(r) u0 = C_Lc_pw; end -function [pl,ql,pr,qr] = bound(rl,cl,rr,cr,t) +function [pl, ql, pr, qr] = bound(rl, cl, rr, cr, t) pl = 0; ql = 1; % assume that the concentration boundary % is the average of the initial and % theoretical final concentration in bulk - pr = cr - (C_Lbf + C_Lbf)/2; + pr = cr - (C_Lb0 + C_Lbf) / 2; qr = 0; end diff --git a/tex/thesis.tex b/tex/thesis.tex index e42dfac..50562c7 100644 --- a/tex/thesis.tex +++ b/tex/thesis.tex @@ -1566,24 +1566,25 @@ the center of the microcarriers was reached. We also assumed that the pores in the microcarriers were large enough that the interactions between the \gls{stp} and surfaces would be small, thus the apparent diffusivity could be represented as a fraction of the diffusion coefficient of \gls{stp} in water. This model was -given by \cref{eqn:stp_diffision_1,eqn:stp_diffision_2}: +given by \cref{eqn:stp_diffusion_1,eqn:stp_diffusion_2}: % TODO actually derive these equations, eg state the initial conditions and % governing equation \begin{equation} - \label{eqn:stp_diffision_1} + \label{eqn:stp_diffusion_1} \frac{dr}{dt} = \frac{-D_{app}C}{Br(1-r/R)} \end{equation} \begin{equation} - \label{eqn:stp_diffision_2} + \label{eqn:stp_diffusion_2} \frac{dC}{dt} = \frac{-4 \pi n D_{app} C}{V(1/r-1/R)} \end{equation} \noindent where \begin{itemize}[label={}] \item $D_{app}$ is the apparent diffusion rate which is equal to $D_{STP}\beta$ -\item $D_{STP}$ the diffusion rate of \gls{stp} in water +\item $D_{STP}$ the diffusion rate of \gls{stp} (or \glspl{mab} for later + calculations) in water \item $\beta$ a fractional parameter representing the tortuousity and void fraction of the microcarriers (here called the `geometric diffusivity') \item $r$ is the interfatial radius of the unbound biotin within a microcarrier @@ -1602,11 +1603,60 @@ equations were then used analogously to describe the reaction profile of \glspl{mab} assuming a diffusion rate of \SI{4.8e-7}{\cm\squared\per\second}\cite{Sherwood1992}. -These same coefficients -were used in determining the kinetics of the washing steps, and -\SI{5.0e-6}{\cm\squared\per\second}\cite{Niether2020} was used as the diffusion -coefficient for free biotin (which should be the only species left in solution -after all the \gls{snb} has hydrolyzed). +To model the washing of the microcarriers, they once again were assumed to be +porous spheres filled with whatever amount of reagent was left unbound from the +previous step (which was assumed to be equal to concentration in the +supernatent). The diffusion out of the microcarriers is given by the following +partial differential equation and boundary conditions: + +\begin{equation} + \label{eqn:stp_washing} + \frac{\partial C_i}{\partial t} = \frac{1}{r^2}\frac{\partial}{\partial + r}\left(r^2 D_{app} \frac{\partial C_i}{\partial r}\right) +\end{equation} + +\begin{equation} + \label{eqn:stp_washing_left_bc} + C_i(r, 0) = C_{i,0} +\end{equation} + +\begin{equation} + \label{eqn:stp_washing_left_bc} + N_i(0, t) = 0 +\end{equation} + +\begin{equation} + \label{eqn:stp_washing_right_bc} + C_i(R, t) = (C_{b,0}+C_{b,\infty}) / 2 +\end{equation} + +\noindent where (in addition to the variables given already for +\cref{eqn:stp_diffusion_1,eqn:stp_diffusion_2}) +\begin{itemize}[label={}] +\item $N_i$ is the radial flux of the species in question inside the + microcarriers +\item $C_i$ is the concentration of the species in question inside the + microcarriers +\item $C_{i,0}$ is the initial concentration of the species in question inside + the microcarriers (which is assumed to be the concentration in the bulk before + the wash volume is added) +\item $C_{b,0}$ is the initial bulk concentration of the species in question + outside the microcarriers after the initial wash volume has been added +\item $C_{b,\infty}$ is the final bulk concentration of the species in + question outside the microcarriers +\end{itemize} + +Note that in order to avoid solving a moving boundary value problem, the +concentration at the boundary of the microcarriers was fixed at the average of +the final and initial concentration expected to be observed in bulk. This should +be a reasonable assumption given that the volume inside the microcarriers is +tiny compared to the amount of volume added in the wash, thus the boundary +concentration should change little. + +The same diffusion coefficients were used in determining the kinetics of the +washing steps, and \SI{5.0e-6}{\cm\squared\per\second}\cite{Niether2020} was +used as the diffusion coefficient for free biotin (which should be the only +species left in solution after all the \gls{snb} has hydrolyzed). All diffusion coefficients were taken to be valid at \gls{rt} and in \gls{di} water, which is a safe assumption given that our reaction medium was 1X @@ -1615,8 +1665,6 @@ water, which is a safe assumption given that our reaction medium was 1X See \cref{sec:appendix_binding} and \cref{sec:appendix_washing} for the MATLAB code (and output in the case of the washing steps) used. -% METHOD add the equation governing the washing steps - \subsection{Luminex Analysis}\label{sec:luminex_analysis} Luminex was performed using a \product{ProcartaPlex kit}{\thermo}{custom} for @@ -1910,7 +1958,7 @@ this experimental binding data to fit a continuous model for the \gls{stp} binding reaction. Using the diffusion rate of the \gls{stp} (\SI{6.2e-7}{\cm\squared\per\second}), we then calculated the geometric diffusivity of the microcarriers to be 0.190 (see -\cref{eqn:stp_diffision_1,eqn:stp_diffision_2}). +\cref{eqn:stp_diffusion_1,eqn:stp_diffusion_2}). % RESULT state how I calculated the number of mab/surface area Using this effective diffusivity and the known diffusion coefficient of a