The present invention relates to variant AAV capsid polypeptides, wherein the variant AAV capsid polypeptides exhibit increased transduction and/or tropism in human pancreatic tissue or human islets as compared non-variant parent capsid polypeptides.

Disclosed are methods for performing transcription-dependent directed evolution (TRADE) and novel AAV capsids selected using such methods.

The invention described herein provides a recombinant DNA viral particle comprising a protein shell encapsulating an RNA vector genome, as well as related compositions and uses thereof.

Described herein are methods of generating engineered viral capsid variants. Also described herein are engineered viral capsid variants, engineered viral particles and formulations and cells thereof. Also described herein are vector systems containing an engineered viral capsid polynucleotide and uses thereof.

Closed-ended, linear, duplex (CELiD) DNA molecules, recombinant AAV (rAAV), particles comprising CELiD DNA, methods of making such molecules and particles, and therapeutic applications of such particles.

The invention discloses a method of distinguishing empty and full capsids in a virus preparation or loaded and non-loaded non-viral gene therapy vectors. The method comprises the steps of: a) providing a preparation of viral particles or gene therapy vectors; b) subjecting the preparation to interferometric scattering mass spectrometry (ISCAMS), in an interferometric scattering microscope, to generate mass distribution data for the viral particles; c) determining the levels of empty capsids and capsids comprising a genome among the viral particles or the loaded and non-loaded vectors from the mass distribution data.

The present disclosure relates to methods of producing monoclonal antibodies in animals. In particular, the disclosure provides a method of producing, in vivo, antibodies against viral capsids (VCs) derived from a non-enveloped virus (NEV). The method includes administering to a subject, by hydrodynamic-based transfection, a first set of genetic material encoding NEV structural proteins to induce the subject's intracellular translation and assembly of the proteins into viral capsids. The method also includes administering a second set of genetic material encoding NEV non-structural proteins to facilitate the intracellular assembly of the NEV structural proteins. Thus, this method may be used to produce subject-generated antibody-producing cells that secrete anti-VC antibodies that may be harvested and screened for monoclonal anti-VC antibodies.

Disclosed are methods for performing transcription-dependent directed evolution (TRADE) and novel AAV capsids selected using such methods. This disclosure also provides novel AAV capsid mutants. TRADE technology was used to identify novel AAV vectors that mediate neuronal transduction in the brain following intravenous administration. Application of TRADE in vivo resulted in the identification of new AAV capsids that can transduce neurons more efficiently and more specifically than AAV9 in the brain following administration of the new AAV capsids. The disclosed methods may be used to identify AAV capsids that target various cell populations.

Provided herein are methods and compositions for the separation of an adeno-associated virus (AAV) particle from a mixture of the AAV and at least one contaminant using anion exchange chromatography. These methods and compositions allow for improved purification of complete AAV particles from contaminants such as AAV particles that lack a complete genome (e.g., empty capsids) and AAV degradation products.

The present disclosure describes methods and systems for use in the production of adeno-associated virus (AAV) particles, including recombinant adeno-associated virus (rAAV) particles. In certain embodiments, the production process and system use Spodoptera frugiperda insect cells (such as Sf9 or Sf21) as viral production cells. In certain embodiments, the production process and system use Baculoviral Expression Vectors (BEVs) in the production of AAV particles. In certain embodiments, the production process and system allow for the controlled expression of AAV capsid proteins, such as VP1, VP2 and VP3.