Abstract: The invention features methods and apparatus for producing lipid nanoparticles. Methods of the invention include injecting a lipid solution into an aqueous solution at an automated rate (e.g., a rate controlled by a servo pump). The invention provides methods and apparatus for making lipid nanoparticles possessing a wide range of lipid components and hydrophilic encapsulants, including nucleic acids (e.g., mRNA). Also provided are nanoparticles and compositions thereof made by methods and apparatus of the invention.

Abstract: The invention features methods and apparatus for concentrating a nucleic acid. Methods of the invention include providing an initial suspension of a nucleic acid and an initial liquid, contacting the initial suspension with a housing having a filter that does not pass the nucleic acid, pressurizing the housing to produce a filtrate and a nucleic acid retentate from the initial solution, and detecting the volume of the nucleic acid retentate. Apparatus of the invention include a chamber configured to hold a filter housing containing a nucleic acid suspension, a pressure source to filter the suspension, and a detector to depressurize the housing upon detecting the volume reaching a predetermined threshold. The methods and apparatus described herein are useful in filtering, concentrating, and reconstituting nucleic acids, such as mRNA, in processes such as complete buffer replacement.

Abstract: The invention features methods and apparatus for purifying a sample including an RNA (e.g., an RNA having a poly(A) sequence, such as an mRNA). Methods of the invention involve adding a liquid to a sample container including the sample and a solid affinity material (e.g., oligo(dT) or hydroxyapatite), agitating the contents of the sample, removing the liquid, and eluting the RNA of interest from the sample container. The apparatus of the invention include a sample container and one or more components such as a reservoir, collection container, pressure generating source, valve assembly, and agitation mechanism. Certain apparatus include a rotatable magnet or magnetizable component. The methods and apparatus described herein are useful in providing RNA (e.g., RNA having a poly(A) sequence, such as mRNA) in high yield and with high purity using less solvent than conventional purification techniques.