Julia E Fletcher

US Patent:

20060110788, May 25, 2006

Filed:

Oct 3, 2005

Appl. No.:

11/240651

Inventors:

Wieslaw Kudlicki - Carlsbad CA, US
Shiranthi Keppetipola - Vista CA, US
Julia Fletcher - Vista CA, US
Ashley Getbehead - Oceanside CA, US
Federico Katzen - Carlsbad CA, US
Laura Vozza-Brown - Carlsbad CA, US

Assignee:

Invitrogen Corporation - Carlsbad CA

International Classification:

C12P 21/06
C12N 15/74

US Classification:

435068100, 435320100

Abstract:

Compositions, methods and kits for in vitro systems for synthesis of biomolecules such as polypeptides, are provided herein. Cell extracts that provide enhanced yields of soluble proteins using in vitro protein synthesis methods are provided. The invention also includes methods for producing high yields of proteins by the addition of a feeding solution that includes amino acids and an energy source to an ongoing in vitro synthesis system. The invention also includes methods of using a high-yield in vitro synthesis system to produce large quantities of proteins with incorporated labeled amino acids for analysis by methods such as by NMR. The invention further includes vectors for enhanced production of proteins from nucleic acid templates using in vitro synthesis systems.

US Patent:

20060211083, Sep 21, 2006

Filed:

Jan 20, 2006

Appl. No.:

11/336644

Inventors:

Federico Katzen - Carlsbad CA, US
Wieslaw Kudlicki - Carlsbad CA, US
Julia Fletcher - Carlsbad CA, US

International Classification:

C12P 21/06
C07K 14/47

US Classification:

435068100, 530395000

Abstract:

Provided herein are products and processes for efficiently synthesizing biomolecules in vitro using cell-free extracts derived from mammalian cells and insect cells. In an embodiment, provided is a process for preparing a cell-free extract from insect cells and mammalian cells that efficiently synthesizes post-translationally modified target proteins (e.g., glycosylated target proteins). In some embodiments, a ribonucleic acid is synthesized in vitro that comprises a cap, a 5′ untranslated region comprising an 18S rRNA binding ribonucleotide sequence, and a target ribonucleotide sequence. It has been determined that such ribonucleic acids result in efficient in vitro synthesis of a target protein using cell-free extracts derived from non-rabbit mammalian cells and insect cells.

US Patent:

20070117179, May 24, 2007

Filed:

Sep 27, 2006

Appl. No.:

11/535960

Inventors:

Wieslaw Kudlicki - Carlsbad CA, US
Julia Fletcher - Carlsbad CA, US
Federico Katzen - Carlsbad CA, US

Assignee:

INVITROGEN CORPORATION - Carlsbad CA

International Classification:

C12P 21/06

US Classification:

435068100

Abstract:

In vitro protein synthesis systems and methods are provided that produce membrane proteins in soluble form. In some aspects, the invention provides methods of synthesizing proteins using in vitro protein synthesis systems that include an apolipoprotein, in which higher yields of soluble protein are produced than in the absence of the apolipoprotein. Apolipoproteins useful in the present invention include naturally occurring apolipoproteins, as well as sequence variants of wild-type apolipoproteins, and engineered apolipoproteins. The apolipoproteins can be provided in an in vitro protein synthesis system associated with lipid or not associated with lipid. The invention also provides compositions and kits for synthesis of proteins in soluble form, in which the compositions and kits include cell extracts for protein translation and at least one apolipoprotein biomolecule.

US Patent:

20080248565, Oct 9, 2008

Filed:

Feb 29, 2008

Appl. No.:

12/040798

Inventors:

Federico Katzen - Carlsbad CA, US
Julia Fletcher - Carlsbad CA, US
Wieslaw Kudlicki - Carlsbad CA, US
Joseph Beechem - Eugene OR, US
Lilin Wang - San Diego CA, US

Assignee:

INVITROGEN CORPORATION - Carlsbad CA

International Classification:

C12N 5/06
C07K 14/00

US Classification:

435325, 530359

Abstract:

Systems and methods are provided for producing a protein of interest that is typically not amenable to expression in soluble form in in vitro expression systems. In some aspects, the invention provides methods of synthesizing proteins using in vitro protein synthesis systems that include a scaffold protein such as apolipoprotein or an amphipathic alpha helix containing (“AAHC”) protein, in which higher yields of soluble protein are produced than in the absence of the scaffold protein. The scaffold proteins may be provided in an in vitro protein synthesis system associated with lipid or not associated with lipid. The scaffold protein may be provided as a protein per se or may be encoded by a nucleic acid template and co-expressed with the protein of interest. The invention also provides compositions and kits for synthesis of proteins in soluble form, in which the compositions and kits include cell extracts for protein expression and isolation.

US Patent:

20090161828, Jun 25, 2009

Filed:

Dec 11, 2008

Appl. No.:

12/333191

Inventors:

Federico Katzen - Carlsbad CA, US
Julia Fletcher - Carlsbad CA, US
Wieslaw Kudlicki - Carlsbad CA, US
Joseph Beechem - Eugene OR, US
Lilin Wang - San Diego CA, US

Assignee:

LIFE TECHNOLOGIES - Carlsbad CA

International Classification:

G01N 23/207
C07K 14/00
C12N 9/10
C12N 9/48
C12N 9/16

US Classification:

378 73, 530359, 435193, 435212, 435196

Abstract:

Systems and methods are provided for producing a protein of interest that is typically not amenable to expression in soluble form in in vitro expression systems. In some aspects, the invention provides methods of synthesizing proteins using in vitro protein synthesis systems that include a scaffold protein such as apolipoprotein or an amphipathic alpha helix containing (“AAHC”) protein, in which higher yields of soluble protein are produced than in the absence of the scaffold protein. The scaffold proteins may be provided in an in vitro protein synthesis system associated with lipid or not associated with lipid. The scaffold protein may be provided as a protein per se or may be encoded by a nucleic acid template and co-expressed with the protein of interest. The invention also provides compositions and kits for synthesis of proteins in soluble form, in which the compositions and kits include cell extracts for protein expression and isolation.

US Patent:

20090209032, Aug 20, 2009

Filed:

Sep 15, 2008

Appl. No.:

12/211094

Inventors:

Wieslaw Antoni Kudlicki - Carlsbad CA, US
Shiranthi Keppetipola - Vista CA, US
Julia Fletcher - Carlsbad CA, US
Ashley Elaine Getbehead - Oceanside CA, US
Federico Katzen - Carlsbad CA, US
Laura Vozza-Brown - Carlsbad CA, US

Assignee:

INVITROGEN CORPORATION - Carlsbad CA

International Classification:

C12N 15/74
C07K 2/00

US Classification:

4353201, 530402

Abstract:

Compositions, methods and kits for in vitro systems for synthesis of biomolecules such as polypeptides, are provided herein. Cell extracts that provide enhanced yields of soluble proteins using in vitro protein synthesis methods are provided. The invention also includes methods for producing high yields of proteins by the addition of a feeding solution that includes amino acids and an energy source to an ongoing in vitro synthesis system. The invention also includes methods of using a high-yield in vitro synthesis system to produce large quantities of proteins with incorporated labeled amino acids for analysis by methods such as by NMR. The invention further includes vectors for enhanced production of proteins from nucleic acid templates using in vitro synthesis systems.

US Patent:

20100233782, Sep 16, 2010

Filed:

Oct 9, 2009

Appl. No.:

12/576880

Inventors:

Federico Katzen - Carlsbad CA, US
Julia Fletcher - Carlsbad CA, US
Wieslaw Kudlicki - Carlsbad CA, US
Joseph Beechem - Eugene OR, US
Lilin Wang - San Diego CA, US

Assignee:

LIFE TECHNOLOGIES CORPORATION - Carlsbad CA

International Classification:

C12N 9/10
C07K 14/00
C12N 9/48
C12N 9/16

US Classification:

435193, 530350, 435212, 435196

Abstract:

Systems and methods are provided for producing a protein of interest that is typically not amenable to expression in soluble form in in vitro expression systems. In some aspects, the invention provides methods of synthesizing proteins using in vitro protein synthesis systems that include a scaffold protein such as apolipoprotein or an amphipathic alpha helix containing (“AAHC”) protein, in which higher yields of soluble protein are produced than in the absence of the scaffold protein. The scaffold proteins may be provided in an in vitro protein synthesis system associated with lipid or not associated with lipid. The scaffold protein may be provided as a protein per se or may be encoded by a nucleic acid template and co-expressed with the protein of interest. The invention also provides compositions and kits for synthesis of proteins in soluble form, in which the compositions and kits include cell extracts for protein expression and isolation.

US Patent:

20110104781, May 5, 2011

Filed:

Sep 24, 2010

Appl. No.:

12/890386

Inventors:

FEDERICO KATZEN - CARLSBAD CA, US
JULIA FLETCHER - CARLSBAD CA, US
WIESLAW KUDLICKI - CARLSBAD CA, US
JOSEPH BEECHEM - EUGENE OR, US
LILIN WANG - SAN DIEGO CA, US

Assignee:

LIFE TECHNOLOGIES CORPORATION - CARLSBAD CA

International Classification:

C12N 9/00
C07K 14/00
C12N 5/071
C12N 1/16

US Classification:

435183, 530350, 435367, 435365, 435358, 43525421, 43525422, 43525423

Abstract:

Systems and methods are provided for producing a protein of interest that is typically not amenable to expression in soluble form in in vitro expression systems. In some aspects, the invention provides methods of synthesizing proteins using in vitro protein synthesis systems that include a scaffold protein such as apolipoprotein or an amphipathic alpha helix containing (“AAHC”) protein, in which higher yields of soluble protein are produced than in the absence of the scaffold protein. The scaffold proteins may be provided in an in vitro protein synthesis system associated with lipid or not associated with lipid. The scaffold protein may be provided as a protein per se or may be encoded by a nucleic acid template and co-expressed with the protein of interest. The invention also provides compositions and kits for synthesis of proteins in soluble form, in which the compositions and kits include cell extracts for protein expression and isolation.