Melissa A Gee

US Patent:

6893827, May 17, 2005

Filed:

Sep 1, 2000

Appl. No.:

09/654499

Inventors:

Michelle A. J. Palmer - Arlington MA, US
Melissa Gee - Bedford MA, US
Bonnie Tillotson - Belmont MA, US
Xiao-Jia Chang - Lincoln MA, US

Assignee:

Applera Corporation - Bedford MA

International Classification:

C07K014/705
C07K019/00
C12N015/62
G01N033/567

US Classification:

435 71, 435 72, 435 721, 435 697, 536 234

Abstract:

Methods for detecting G-protein coupled receptor (GPCR) activity; methods of assaying GPCR activity; and methods of screening for GPCR ligands, G-protein-coupled receptor kinase (GRK) activity, and compounds that interact with components of the GPCR regulatory process are described.

US Patent:

7235374, Jun 26, 2007

Filed:

Oct 5, 2004

Appl. No.:

10/959611

Inventors:

Michelle A. J. Palmer - Arlington MA, US
Melissa Gee - Bedford MA, US
Bonnie Tillotson - Belmont MA, US
Xiao-jia Chang - Lincoln MA, US

Assignee:

Applera Corporation - Foster City CA

International Classification:

C07K 14/705
C07K 19/00
C12N 15/62
G01N 33/566

US Classification:

435 72, 435 721, 435 697, 536 234

Abstract:

Methods for detecting G-protein coupled receptor (GPCR) activity; methods for assaying GPCR activity; and methods for screening for GPCR ligands, G-protein-coupled receptor kinase (GRK) activity, and compounds that interact with components of the GPCR regulatory process are described. Included are methods for expanding ICAST technologies for assaying GPCR activity with applications for ligand fishing, and agonist or antagonist screening. These methods include: engineering seronine/threonine phosphorylation sites into known or orphan GPCR open reading frames in order to increase the affinity of arrestin for the activated form of the GPCR or to increase the reside time of arrestin on the activated GPCR; engineering mutant arrestin proteins that bind to activated GPCRs in the absence of G-protein coupled receptor kinases which may be limiting; and engineering mutant super arrestin proteins that have an increased affinity for activated GPCRs with or without phosphorylation. These methods are intended to increase the robustness of the GPCR/ICAST technology in situations in which G-protein coupled receptor kinases are absent or limiting, or in which the GPCR is not efficiently down-regulated or is rapidly resensitized (thus having a labile interaction with arrestin). Included are also more specific methods for using ICAST complementary enzyme fragments to monitor GPCR homo- and hetero-dimerization with applications for drug lead discovery and ligand and function discovery for orphan GPCRs.

US Patent:

20020028433, Mar 7, 2002

Filed:

Jan 16, 2001

Appl. No.:

09/759152

Inventors:

Michelle Palmer - Arlington MA, US
Melissa Gee - Bedford MA, US
Bonnie Tillotson - Belmont MA, US
Xiao-Jia Chang - Lincoln MA, US

International Classification:

C12Q001/00
C12N005/06

US Classification:

435/004000, 435/325000

Abstract:

Methods for detecting G-protein coupled receptor (GPCR) activity; methods for assaying GPCR activity; and methods for screening for GPCR ligands, G-protein-coupled receptor kinase (GRK) activity, and compounds that interact with components of the GPCR regulatory process are described. Included are methods for expanding ICAST technologies for assaying GPCR activity with applications for ligand fishing, and agonist or antagonist screening. These methods include: engineering seronine/threonine phosphorylation sites into known or orphan GPCR open reading frames in order to increase the affinity of arrestin for the activated form of the GPCR or to increase the reside time of arrestin on the activated GPCR; engineering mutant arrestin proteins that bind to activated GPCRs in the absence of G-protein coupled receptor kinases which may be limiting; and engineering mutant super arrestin proteins that have an increased affinity for activated GPCRs with or without phosphorylation. These methods are intended to increase the robustness of the GPCR/ICAST technology in situations in which G-protein coupled receptor kinases are absent or limiting, or in which the GPCR is not efficiently down-regulated or is rapidly resensitized (thus having a labile interaction with arrestin). Included are also more specific methods for using ICAST complementary enzyme fragments to monitor GPCR homo- and hetero-dimerization with applications for drug lead discovery and ligand and function discovery for orphan GPCRs.

US Patent:

20120100566, Apr 26, 2012

Filed:

Mar 1, 2010

Appl. No.:

13/254154

Inventors:

Brooks Edwards - Cambridge MA, US
Melissa Gee - Littleton MA, US
Zhixian Wang - Winchester MA, US
Kathleen Skaare - Newton MA, US

Assignee:

LIFE TECHNOLOGIES CORPORATION - Carlsbad CA

International Classification:

C12Q 1/26
C07D 409/04
C07D 323/00

US Classification:

435 25, 549332, 549 51

Abstract:

Chemiluminescent compositions, methods, assays and kits for oxidative enzymes are described. Further disclosed are dioxetane compounds of the form: 0-0 ΛR R R T (i) where R can independently be any branched alkyl or cycloalkyl group which provides stabilization for the dioxetane or where both R groups together form a cycloalkyl or polycycloalkyl moiety spiro bound to the dioxetane ring, wherein each R group or the spiro bound moiety can be unsubstituted or substituted with one or more electron-withdrawing groups or electron donating groups, or groups providing preferential oxidative isozyme substrate recognition, and wherein Ri is an aryl group, or an alkyl group of 1-20 carbon atoms, which can be optionally substituted with 1 or more halogen atoms, and wherein T is an aryl or heteroaryl ring capable of emitting light upon enzyme activated decomposition of the dioxetane I. Kits, methods and assays are also disclosed that comprise the dioxetane compounds.

US Patent:

20130196325, Aug 1, 2013

Filed:

Jul 6, 2011

Appl. No.:

13/808922

Inventors:

Alison Sparks - North Andover MA, US
Zhixian Wang - Winchester MA, US
Melissa Gee - Littleton MA, US

Assignee:

LIFE TECHNOLOGIES CORPORATION - Carlsbad CA

International Classification:

G01N 33/58
C12Q 1/42
C12Q 1/68
C12Q 1/34

US Classification:

435 611, 435 18, 435 21, 435 792

Abstract:

Methods for generating a chemiluminescent enzyme substrate in situ, in aqueous or other assay conditions. Also disclosed are methods to use the substrates to generate light, detect and/or quantify enzymes, antigens, and/or nucleic acids. Kits relating to these methods are also disclosed.

US Patent:

20180299456, Oct 18, 2018

Filed:

Jun 25, 2018

Appl. No.:

16/017824

Inventors:

- Carlsbad CA, US
Zhixian Wang - Winchester MA, US
Melissa Gee - Littleton MA, US

International Classification:

G01N 33/58
C07D 321/00
C12Q 1/42
C12Q 1/34
C07H 15/203
C12Q 1/6816
C07F 9/655
C07F 9/6541
C07F 9/40
C07F 9/12
C07F 9/6558

Abstract:

Methods for generating a chemiluminescent enzyme substrate in situ, in aqueous or other assay conditions. Also disclosed are methods to use the substrates to generate light, detect and/or quantify enzymes, antigens, and/or nucleic acids. Kits relating to these methods are also disclosed.

US Patent:

20170137862, May 18, 2017

Filed:

Nov 7, 2016

Appl. No.:

15/345098

Inventors:

- Carlsbad CA, US
Brooks EDWARDS - Cambridge MA, US
Melissa GEE - Bedford MA, US
Zhixian WANG - Bedford MA, US
Kathleen SKAARE - W. Newton MA, US

International Classification:

C12Q 1/26

Abstract:

Chemiluminescent compositions, methods, assays and kits for oxidative enzymes are described. Further disclosed are dioxetane compounds of the form:where R can independently be any branched alkyl or cycloalkyl group which provides stabilization for the dioxetane or where both R groups together form a cycloalkyl or polycycloalkyl moiety spiro bound to the dioxetane ring, wherein each R group or the spiro bound moiety can be unsubstituted or substituted with one or more electron-withdrawing groups or electron donating groups, or groups providing preferential oxidative isozyme substrate recognition, and wherein Ris an aryl group, or an alkyl group of 1-20 carbon atoms, which can be optionally substituted with 1 or more halogen atoms, and wherein T is an aryl or heteroaryl ring capable of emitting light upon enzyme activated decomposition of the dioxetane I.Kits, methods and assays are also disclosed that comprise the dioxetane compounds.

US Patent:

20150284766, Oct 8, 2015

Filed:

Apr 23, 2015

Appl. No.:

14/694758

Inventors:

- Carlsbad CA, US
Brooks Edwards - Cambridge MA, US
Melissa GEE - Littleton MA, US
Zhixian WANG - Winchester MA, US
Kathleen SKAARE - Newton MA, US

International Classification:

C12Q 1/26
C07D 409/04
C07D 323/00

Abstract:

Chemiluminescent compositions, methods, assays and kits for oxidative enzymes are described. Further disclosed are dioxetane compounds of the form:where R can independently be any branched alkyl or cycloalkyl group which provides stabilization for the dioxetane or where both R groups together form a cycloalkyl or polycycloalkyl moiety spiro bound to the dioxetane ring, wherein each R group or the spiro bound moiety can be unsubstituted or substituted with one or more electron-withdrawing groups or electron donating groups, or groups providing preferential oxidative isozyme substrate recognition, and wherein Ris an aryl group, or an alkyl group of 1-20 carbon atoms, which can be optionally substituted with 1 or more halogen atoms, and wherein T is an aryl or heteroaryl ring capable of emitting light upon enzyme activated decomposition of the dioxetane I.Kits, methods and assays are also disclosed that comprise the dioxetane compounds.