deletions | additions       diff --git a/Method.tex b/Method.tex  index 15f6577..093981c 100644  --- a/Method.tex  +++ b/Method.tex 
...
\subsection{Reagents}  Ranibizumab was purchased from Novartis (Frimley, UK) (Lucentis\textregistered (Lucentis{\textregistered}  10~mg/ml) and bevacizumab from Roche (Welwyn Garden City, UK) (Avastin\textregistered (Avastin{\textregistered}  25~mg/ml). BSA was purchased from Sigma (Poole, UK) at the highest available purity. Aflibercept was purchased from Bayer plc (Newbury, UK) as a 40~mg/ml solution (Eylea\textregistered). All other reagents were of the highest available purity and were either from Sigma (Poole, UK), VWR (Lutterworth, UK), or Merck (Hoddesdon, UK) unless otherwise specified. \subsection{Sample preparation}  All protein solutions were diluted with sterile phosphate buffered saline (PBS; Sigma; pH~7.4) to 5mg/mL and then dialyzed twice against phosphate buffered saline (PBS) pH 7.4 using D-Tube Midi Dialyzer units from Novagen (6-8~kD cut-off). Subsequently, the purity of the dialysed proteins was confirmed by standard SDS-PAGE. The proteins were then conjugated to the ruthenium dye. This was achieved by reacting the succinimidyl ester-modified dye, Bis(2,2′-bipyridine)-4′-methyl-4-carboxybipyridine-ruthenium N-succinimidyl ester-bis(hexafluorophosphate) (synonym, Ru(bpy)$_2$(mcbpy-O-Su-ester)(PF$_6$)$_2$, Sigma-Aldrich) by using a succinimidyl ester-modified fluorophore with a short linker (Invitrogen, F6130). Conjugation reactions were performed in PBS adjusted with bicine buffer to pH 8.6 at 2~mg/ml protein concentration with the activated fluorophore ester being used in excess (2-fold and 3.5-fold for proteins $>$60~kD and ranibizumab, respectively). The reaction was stopped after two hours and conjugated proteins were separated from remaining free dye by size exclusion chromatography (two times via 7~kD cut-off Zeba\textregistered Zeba{\textregistered}  spin columns (Thermo Fisher, UK)). Using this method we generated the various proteins conjugated to the fluorophore in the following dye:protein ratios: ranibizumab 1.1:1; BSA 1.2:1; aflibercept 1.8:1; bevacizumab 1.3:1. Azide (2.0~mM) was added to all dye-conjugated drugs to protect them from microbial deterioration. \subsection{Anisotropy measurements}