Protein arrays and methods of using and making the same

• US 10,457,935 B2
• Filed: 11/10/2011
• Issued: 10/29/2019
• Est. Priority Date: 11/12/2010
• Status: Active Grant

• Current Assignee: Gen9, Inc. (Ginkgo Bioworks Holdings, Inc.)
• Original Assignee: Gen9, Inc. (Ginkgo Bioworks Holdings, Inc.)
• Inventors: Jacobson, Joseph, Hudson, Michael E., Chu, Larry Li-Yang
• Primary Examiner(s): Zhang, Kaijiang
• Application Number:
  US13/884,446
• Publication Number:
  US 20130296192A1
• Time in Patent Office: 2,910 Days
• Field of Search: None
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00509 Microcolumns
• CPC Class Codes: B01J 2219/00531 essentially square
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00644 the porous medium being present in discrete locations, e.g. gel pads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: C12N 15/1086 Preparation or screening of expression libraries, e.g. reporter assays
• CPC Class Codes: C40B 50/14 Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support

Abstract

Methods and devices are provided for preparing a protein array having a plurality of proteins. In one embodiment, the method includes providing a plurality of nucleic acids each having a predefined sequence and expressing in vitro a plurality of proteins from the plurality of nucleic acids. In another embodiment, protein arrays having a solid surface and a microvolume are also provided. The solid surface can have a plurality of anchor oligonucleotides capable of hybridizing with a plurality of nucleic acids. The microvolume can cover each of the plurality of anchor oligonucleotides and can be configured to produce a polypeptide from each of the plurality of nucleic acids.

Claims

1. A method for preparing a protein array having a plurality of proteins, the method comprising:

(a) providing a plurality of nucleic acids each having a predefined sequence and each encoding a protein of interest, wherein the plurality of nucleic acids are immobilized on discrete features of a surface; and

(b) expressing in vitro a plurality of proteins of interest from the plurality of nucleic acids, wherein the plurality of proteins are each present in solution at the discrete features on the surface;

wherein each of the plurality of proteins of interest is expressed in a self-contained volume having a size of 1 picoliter to 1 nanoliter.

2. The method of claim 1, further comprising (c) measuring an activity of each of the plurality of proteins that are present in solution.

3. The method of claim 1, wherein the plurality of nucleic acids are synthesized on a solid surface.

4. The method of claim 1, wherein each of the plurality of nucleic acids comprises a regulatory genetic sequence.

5. The method of claim 1, wherein each of the plurality of proteins of interest is expressed in vitro in a well of a micro-well plate.

6. The method of claim 1, wherein each of the plurality of proteins of interest is expressed in vitro at a different feature of a solid surface.