Barcoded Beads And Method For Making The Same By Split-Pool Synthesis
Patent No. US11932902 (titled "Barcoded Beads And Method For Making The Same By Split-Pool Synthesis") was filed by Roche Sequencing Solutions Inc on Aug 11, 2023.
What is this patent about?
’902 is related to the field of cellular barcoding and single-cell analysis. The invention addresses the need for methods to identify and quantify target molecules within individual cells of a complex population, while retaining information about the cell of origin. Existing methods often require significant amounts of biological samples or lack cell-specific information, hindering the discovery of new therapeutic and diagnostic targets.
The underlying idea behind ’902 is to create a unique identifier, or cell origination barcode (COB) , for each cell in a population. This is achieved by iteratively appending pre-made oligonucleotides to beads in a split-pool process. Each round of splitting and oligonucleotide addition creates a more complex barcode, allowing for a large number of unique identifiers to be generated.
The claims of ’902 focus on a method for adding cell origination barcodes onto beads. This involves splitting a pool of beads into multiple reaction volumes, appending different pre-made oligonucleotides to the beads in each volume, and then pooling the beads. This process is repeated to build up the COBs, with each oligonucleotide addition contributing to the final barcode sequence. The independent claims also cover a population of beads with different COBs.
In practice, the method involves attaching the beads to cells or cellular components. The pre-made oligonucleotides, or assayable polymer subunits (APSs) , are designed to link sequentially, building up a unique barcode for each cell. This allows for the identification and quantification of target molecules within each cell, as well as the ability to trace the origin of those molecules back to the specific cell they came from.
This approach differs from prior solutions by creating barcodes in a stepwise manner using a split-pool process. This allows for the generation of a large number of unique barcodes without the need for complex synthesis or sequencing steps. The use of pre-made oligonucleotides also simplifies the process and makes it more scalable, enabling the analysis of large cell populations with high accuracy and sensitivity. The split-pool synthesis is key to generating diverse barcodes.
How does this patent fit in bigger picture?
Technical landscape at the time
In the early 2010s when ’902 was filed, methods for analyzing gene expression and protein quantification in single cells were emerging, but at a time when multiplexing these measurements was still challenging. At that time, single-cell analysis at a high throughput was not trivial, and methods commonly relied on isolating individual cells before analysis, when hardware or software constraints made complex barcoding strategies non-trivial.
Novelty and Inventive Step
The examiner approved the claims because the prior art did not teach or suggest using pre-made oligonucleotide tags in a split-pool approach for cell origination barcoding. While the closest prior art taught split-pool DNA synthesis on beads for barcoding, it used a DNA synthesizer to add bases one-by-one, rather than assembling larger cell origination barcodes from pre-made oligonucleotides.
Claims
This patent contains 30 claims, of which claims 1, 17, and 23 are independent. Independent claims 1 and 17 are directed to methods for adding cell origination barcodes onto beads using a split-pool process. Independent claim 23 is directed to a population of beads comprising cell origination barcodes. The dependent claims generally elaborate on and refine the methods and compositions of the independent claims.
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Description
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US11932902
- Application Number
- US18233214
- Filing Date
- Aug 11, 2023
- Status
- Granted
- Expiry Date
- Jan 31, 2032
- External Links
- Slate, USPTO, Google Patents