# US 10,308,931 B2 · Methods for screening proteins using DNA encoded chemical libraries as templates for enzyme catalysis

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- Number: US 10,308,931 B2
- Document ID: US10308931B2
- Status: Granted
- Kind: B2
- Application number: 14/840,551
- Filed: 2015-08-31
- Published: 2015-12-17
- Granted: 2019-06-04
- Earliest claimed: 2012-03-21
- Assignees: Gen9, Inc.
- Applicants: Gen9, Inc.
- Inventors: Michael E. Hudson; Joseph Jacobson
- Web page: https://patentgrep.com/patent/US10308931B2

## CPC

- C12N 15/1034: {Isolating an individual clone by screening libraries}
- C12N 15/1037: {Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display}
- C12N 15/1086: {Preparation or screening of expression libraries, e.g. reporter assays}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Disclosed are methods, compositions and devices for screening a protein library for proteins having a desired activity, such as capable of catalyzing the formation of a bond between two reactants. In an exemplary embodiments, a plurality of proteins are expressed in vitro from a plurality of nucleic acids, the plurality of proteins are exposed with two single stranded oligonucleotides having complementary sequences, each oligonucleotide having a reactant and a fluorophore, the fluorescence of the protein-reactant-oligonucleotide-fluorophore complexes is detected and the complexes showing detectable fluorescence energy transfer are isolated, thereby isolating proteins having the desired enzymatic activity.

## Claims

### Claim 1

A method for screening a protein library for a protein having a desired activity, the method comprising:
a) providing, at distinct features of a solid support, a plurality of nucleic acids, each nucleic acid having a predefined sequence;
b) expressing in vitro a plurality of proteins from the plurality of nucleic acids;
c) exposing the plurality of proteins to a first and a second oligonucleotide each, at one end, having a first and a second reactant, respectively, wherein a protein having a desired activity binds to the first and second reactants, thereby forming a complex; and
d) ligating free ends of the first and second oligonucleotides complexed with the protein having the desired activity.

### Claim 2 (depends on 1)

The method of claim 1 wherein the first and second oligonucleotides are double-stranded oligonucleotides.

### Claim 3 (depends on 1)

The method of claim 1 wherein the first and second oligonucleotides are single-stranded oligonucleotides.

### Claim 4 (depends on 3)

The method of claim 3 further comprising adding a helper oligonucleotide capable of bridging the single-stranded oligonucleotides bound to the protein having the desired activity.

### Claim 5 (depends on 1)

The method of claim 1 wherein each distinct feature is a well of a micro well plate.