Abstract: Methods discussed herein are directed to polymerase chain reaction (PCR) techniques, and more specifically quantitative-discrete PCR, wherein individual amplification reactions are performed on a per-payload basis among singly-captured and singly-isolated vesicles, for instance within individually sealed microwell reactors. These techniques enable post-process quality control measurements of per-vesicular manufacture loading efficiency and encapsulation efficiency of nucleic acid active ingredients in vesicular biologics, employing quantitative-discrete PCR techniques. By measuring variability of vesicular encapsulation of nucleic acids at a per-vesicular manufacture level of granularity, such techniques can enable collection of data that may be used to perform post-process formulation upon vesicular biologics, and to yield more homogenous formulations from both synthetic and biogenesis pathways.

Abstract: Described herein are systems using injectionless gel electrophoresis (GE), such as thermal GE (TGE), to selectively separate, concentrate, quantify, and/or otherwise analyze target analytes. Inline preconcentration and separation are demonstrated to resolve analytes, exemplified by resolving double-stranded miRNA-probe hybrids from excess single-stranded probes and analyzing multiple conformations of a protein. Microfluidic devices having a tapered channel are described, which improve detection sensitivity and separation resolution. The described separation strategy and microfluidic device designs establish injectionless gel electrophoresis as a simple, low-cost analysis method, for instance for analyzing clinical and pharmaceutical samples, including for miRNA, protein, and other biomolecular analyses.