Randomly-Branched Polymers by Size Exclusion Chromatography with Triple Detection: Computer Simulation Study for Estimating Errors in the Distribution of Molar Mass and Branching Degree

2013 ◽  
Vol 23 (2) ◽  
pp. 90-100 ◽  
Author(s):  
Luis A. Clementi ◽  
Jorge R. Vega ◽  
Gregorio R. Meira
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Nicole L. McIntosh ◽  
Geoffrey Y. Berguig ◽  
Omair A. Karim ◽  
Christa L. Cortesio ◽  
Rolando De Angelis ◽  
...  

AbstractAdeno associated virus (AAV) capsids are a leading modality for in vivo gene delivery. Complete and precise characterization of capsid particles, including capsid and vector genome concentration, is necessary to safely and efficaciously dose patients. Size exclusion chromatography (SEC) coupled to multiangle light scattering (MALS) offers a straightforward approach to comprehensively characterize AAV capsids. The current study demonstrates that this method provides detailed AAV characterization information, including but not limited to aggregation profile, size-distribution, capsid content, capsid molar mass, encapsidated DNA molar mass, and total capsid and vector genome titer. Currently, multiple techniques are required to generate this information, with varying accuracy and precision. In the current study, a new series of equations for SEC-MALS are used in tandem with intrinsic properties of the capsids and encapsidated DNA to quantify multiple physical AAV attributes in one 20-min run with minimal sample manipulation, high accuracy, and high precision. These novel applications designate this well-established method as a powerful tool for product development and process analytics in future gene therapy programs.


e-Polymers ◽  
2005 ◽  
Vol 5 (1) ◽  
Author(s):  
Martina Adler ◽  
Harald Pasch ◽  
Christian Meier ◽  
Raimund Senger ◽  
Hans-Günter Koban ◽  
...  

AbstractA robust and reproducible method for the molar mass analysis of cationic copolymers based on dimethylaminoethyl methacrylate or trimethylammonioethyl methacrylate and different (meth)acrylates has been developed. Size exclusion chromatography (SEC) using a novel polyester-based packing as the stationary phase and dimethylacetamide (DMAC) as the mobile phase yields highly accurate results for copolymers with an amino comonomer content up to 50 wt.-%. To suppress the different polar and ionic interactions between sample molecules, stationary phase and eluent, DMAC was modified with LiBr and tris(hydroxymethylamino) methane (TRIS). Calibrating the SEC system with poly(methyl methacrylate) of narrow polydispersity, molar masses were obtained that are in good agreement with viscosity data. Reproducibility and robustness of the novel method were proven by running samples for an extended period of two weeks.


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