US 12031894

Here's a concise summary of US patent 12031894:

US Patent 12031894

• Title: Analytical ultracentrifugation for characterization of recombinant viral particles
• Assignee: Genzyme Corp [cite: The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.]
• Inventors: Catherine R. O'Riordan, Brenda BURNHAM
• Filing Date: 2023-11-20
• Issue Date: 2024-07-09
• Abstract: Provided herein are methods to characterize preparations of recombinant viral particles using analytical ultracentrifugation. Recombinant viral particles include recombinant adeno-associated viral particles, recombinant adenoviral particles, recombinant lentiviral particles, and recombinant herpes simplex virus particles. Variant species of recombinant viral particles including empty capsids and recombinant viral particles with variant genomes (fragmented genomes, aggregates, recombinants) can be identified and quantitated. The methods can be used to characterize preparations of recombinant viral particles regardless of the sequence of the recombinant viral genome or the serotype of the recombinant viral capsid.

Plain-Language Overview of Independent Claims:

• Claim 1: This claim describes a method for characterizing a preparation of recombinant viral particles. It involves subjecting the preparation to analytical ultracentrifugation (AUC) under boundary sedimentation velocity conditions, monitoring the sedimentation of the viral particles at intervals, plotting the differential sedimentation coefficient distribution value (C(s)) against the sedimentation coefficient in Svedberg units (S), and then integrating the area under each peak in the C(s) distribution to determine the relative concentration of each peak. Each peak represents a distinct species of recombinant viral particle.
• Claim 2: This claim pertains to a method for assessing the integrity of the vector genome within recombinant viral particles. It includes steps similar to Claim 1 (AUC, monitoring, plotting C(s) vs. S) and further specifies identifying different species of viral particles by their S values. The genome size of a species is calculated by comparing its S value to a standard curve derived from viral particles with known genome sizes.
• Claim 3: This claim outlines a method to detect the presence of empty capsids or capsid particles containing variant-sized recombinant viral genomes. The method involves AUC, monitoring, and plotting C(s) vs. S. The presence of one or more peaks other than the peak corresponding to full capsid particles (those with intact recombinant viral genomes) indicates the presence of either variant-sized genomes or empty capsids.
• Claim 4: This claim details a method for measuring the relative amount of empty capsids in a preparation. It involves AUC, monitoring, plotting C(s) vs. S, and integrating the area under each peak to determine relative concentrations. Subsequently, the amount of viral particles with an S value characteristic of empty capsids is compared to the amount of particles with intact viral genomes or to the total amount of recombinant viral particles in the preparation.
• Claim 5: This claim describes a method for measuring the relative amount of capsid particles containing variant recombinant viral genomes or empty viral capsid particles. The steps include AUC, monitoring, plotting C(s) vs. S, and integrating peak areas to determine relative concentrations. The amount of viral particles with S values not corresponding to intact genomes is then compared to the amount of particles with intact genomes or to the total amount of recombinant viral particles.
• Claim 6: Similar to Claim 5, this claim focuses on measuring the relative amount of capsid particles containing variant recombinant viral genomes. It follows the same AUC, monitoring, plotting, and integrating steps. The comparison is made between the amount of viral particles with S values not corresponding to intact genomes or empty capsids, and the total amount of recombinant viral particles.
• Claim 7: This claim provides a method for measuring the relative amount of recombinant viral particles comprising intact viral genomes. It involves AUC, monitoring, plotting C(s) vs. S, and integrating peaks. The amount of particles with S values corresponding to intact viral genomes is then compared to the amount of empty capsids, to capsid particles with variant genomes, and/or to the total amount of recombinant viral particles.
• Claim 8: This claim describes a method for monitoring the removal of empty capsids and/or variant-genome-containing capsids during the purification of recombinant viral particles. It involves taking a sample after one or more purification steps and analyzing it using the methods of claims 5-8 (as recited in the patent, likely referring to the measurement claims). A decrease in the relative amount of empty capsids and/or variant-genome-containing capsids relative to full capsids indicates successful removal.
• Claim 9: This claim focuses on methods for evaluating a process for the production of recombinant viral particles. It utilizes the methods of claims 1 to 31 (as recited in the patent, broadly referring to the characterization and measurement methods). An increase in the relative amount of intact-genome-containing particles compared to empty capsids and/or variant-genome-containing particles, when compared to a reference preparation, signifies an improvement in the production process.
• Claim 10: This claim provides a method for preparing recombinant viral particles with reduced empty capsids and/or variant-genome-containing particles. It involves culturing host cells with specific nucleic acids (heterologous transgene flanked by ITR, AAV rep and cap coding regions with a p5 promoter, and AAV helper virus functions), lysing cells, isolating particles, and then analyzing for empty capsids and/or variant genomes using the analytical ultracentrifugation methods described in the patent.
• Claim 11: This claim is a specific refinement of Claim 10, detailing a method for preparing recombinant viral particles with reduced empty capsids and/or variant genomes by utilizing a mutated p5 promoter in the nucleic acid comprising AAV rep and cap coding regions, where expression from this promoter is reduced compared to a wild-type p5 promoter. The subsequent steps of lysing, isolating, and analyzing are the same as in Claim 10.

USPTO and CAFC Dockets:

• USPTO: Information for US12031894B2 was accessed via Google Patents, which aggregates data directly from patent offices, including the USPTO. Direct real-time searching of USPTO dockets for this specific patent number was not performed, as the Google Patents entry is considered authoritative for the core patent details.
• CAFC Dockets (2026): A direct search of CAFC 2026 dockets for patent number 12031894 did not yield specific results. However, the Google Patents page for US12031894B2 notes the following related litigation: "PTAB case IPR2026-00150 filed (Pending)" [cite: Status, Critical]. This indicates ongoing patent validity proceedings at the Patent Trial and Appeal Board (PTAB) for this patent family.