Multi-well system

• US 8,309,035 B2
• Filed: 04/03/2009
• Issued: 11/13/2012
• Est. Priority Date: 04/05/2008
• Status: Active Grant

• Current Assignee: Single Cell Technology, Inc.
• Original Assignee: Single Cell Technology, Inc.
• Inventors: Chen, Chun-Nan, Jorgensen, Richard Aleck, Bowlby, James O. Jr.
• Primary Examiner(s): LAM, ANN Y
• Application Number:
  US12/418,520
• Publication Number:
  US 20090280992A1
• Time in Patent Office: 1,320 Days
• Field of Search: None
• US Class Current: 422/407
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C12N 15/1065 Preparation or screening of tagged libraries, e.g. tagged microorganisms by STM-mutagenesis, tagged polynucleotides, gene tags
• CPC Class Codes: Y02A 90/10 Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation

Abstract

A multi-well system is described which is comprised of a tray comprising a plurality of wells which may include thousands, tens of thousands or even hundreds of thousands of wells or more. Each of the wells has a unique address. The system includes a tray top comprised of a plurality of areas wherein each of the areas corresponds uniquely to each of the wells and includes a unique address. The system may include a second tray top also comprised of a plurality of areas with unique addresses which uniquely correspond to each of the wells. The areas on the tops have compounds bound to those areas which compounds bind to components present in the wells and are used to obtain specific material in each well for analysis.

Claims

1. A multi-well system for characterizing cells, comprising:

a tray comprising a plurality of wells, each of the wells comprising a side wall, a bottom and an open top;

a first tray top comprising a plurality of areas, each of the areas corresponding uniquely with each well of the tray;

a protein binding agent bound to each of the plurality of areas of the first tray top; and

a second tray top comprising a plurality of areas, each of the areas corresponding uniquely with each well of the tray, wherein there are oligonucleotide tags bound to the plurality of areas of the second tray top, and wherein the oligonucleotide tags are specific tags that correlate nucleic acids obtained from cells in the wells with a physical location of the wells, are specific to the well of interest and thereby allow pooling of the nucleic acids obtained from cells in the wells for subsequent analysis by ultra-high throughput sequencing.

2. The multi-well system of claim 1, wherein the tray is comprised of 1,000 or more wells fabricated at a density greater than 100 wells per cm².

3. The multi-well system of claim 1, wherein the protein binding agent is an antibody binding agent and the antibody binding agent is selected from the group consisting of Protein A, Protein G, Protein L, Protein A/G, and anti-IgG Fcγ

subclass-specific antibodies.

4. The multi-well system of claim 1, further comprising:

an antibody-producing cell bound to a plurality of the plurality of wells in the tray.

5. The multi-well system of claim 1, further comprising:

a third tray top comprising a plurality of areas, each of the areas corresponding uniquely with each well of the tray; and

a protein binding agent bound to each of the plurality of areas of the third tray top.

6. The multi-well system of claim 4, further comprising:

instructions for using the multi-well system to obtain information from antibodies that bind to areas on the first tray top and correlating sequence information from the nucleic acids obtained from the second tray top to the antibody information.

7. The multi-well system of claim 1, further comprising:

a plurality of addresses on the tray wherein a unique address is assignable to each well; and

wherein 10,000 or more wells are fabricated at a density of 200 wells per cm² or more.

8. The multi-well system of claim 1, wherein the tray comprises polydimethylsiloxane (PDMS).

9. The multi-well system of claim 1, wherein the tray is comprised of 1,000 or more wells wherein each well holds a volume of from 10 picoliters to 0.1 milliliters.

10. The multi-well system of claim 1, wherein the nucleic acids obtained from cells in the wells encode light and heavy chain variable regions or portions thereof large enough to construct an antibody.

11. A multi-well system, comprising:

a tray comprised of a frame and a plurality of wells wherein a plurality of the plurality of the wells has bound to its surface an antibody-producing cell;

a first tray top comprising a plurality of areas, each of the areas corresponding uniquely with each of the wells of the tray;

an antibody binding agent bound to each of the plurality of areas of the first tray top;

a second tray top comprising a plurality of areas, each of the areas corresponding uniquely with each well of the tray; and

oligonucleotide tags bound to each of the plurality of areas of the second tray top, wherein the oligonucleotide tags are specific tags that correlate nucleic acids obtained from cells in the wells to a physical location of the wells, are specific to the well of interest and thereby allow pooling of the nucleic acids obtained from the wells for subsequent analysis by ultra-high throughput sequencing.

12. The multi-well system of claim 11, wherein the oligonucleotide tags selectively bind mRNAs which encode antibody light chains or antibody heavy chains.

13. The multi-well system as claimed in claim 12, wherein each of the areas comprise oligonucleotide tags that bind mRNAs that encode antibody light chains and antibody heavy chains.

14. A device to allow optical inspection of cells in a multi-well system, where the multi-well system comprises:

a tray comprising a plurality of wells, each of the wells comprising a side wall, a bottom and an open top;

a first tray top comprising a plurality of areas, each of the areas corresponding uniquely with each well of the tray;

an antibody binding agent bound to each of the plurality of areas of the first tray top;

a second tray top comprising a plurality of areas, each of the areas corresponding uniquely with each well of the tray; and

an oligonucleotide tag bound to each of the plurality of areas of the second tray top, wherein the oligonucleotide tags are specific tags that correlate the nucleic acids obtained from cells in the wells with a physical location of the wells, are specific to the well of interest and thereby allow pooling of nucleic acids obtained from the wells for subsequent analysis by ultra-high throughput sequencing, wherein the device is capable of aligning the tray and the first tray top and the tray and the second tray top.