University of California

Aurora Biosciences Corporation

Anaptysbio, Inc.

University of Oregon

Other

Life Technologies Corporation

Andrew B Cubitt

Roger Y Tsien

Roger Heim

S James Remington

Robert A Horlick

Peter M Bowers

Mats F Ormo

Michael A Whitney

Brian Pollok

Jeffrey Stack

Mats F Orm&ouml;

Robert M Dickson

William E Moerner

Mats F Ormö

This invention relates to methods for the generation of humanized antibodies, particularly a humanized antibody heavy chain protein and a humanized antibody light chain protein. The method comprises using cells that express or can be induced to express Activation Induced Cytidine Deaminase (AID).

Methods of generating libraries and uses thereof

This invention provides tandem fluorescent protein construct including a donor fluorescent protein moiety, an acceptor fluorescent protein moiety and a linker moiety that couples the donor and acceptor moieties. The donor and acceptor moieties exhibit fluorescence resonance energy transfer which is eliminated upon cleavage. The constructs are useful in enzymatic assays.

Method for determining the presence of an enzyme in a sample

This invention relates to methods for the generation of polynucleotide seed libraries and the use of these libraries in generating novel mutants of recombinant proteins and, more particularly, for generating focused libraries of recombinant human antibodies and screening for their affinity binding with target antigens.

This invention is directed towards methods of destabilizing proteins in living cells, and their use for the development of novel assays. In one embodiment, the invention comprises the use of non-cleavable multimerized ubiquitin fusion proteins to destabilize a target protein, such as a reporter moiety. In one aspect of this method the constructs also comprises a linker that operatively couples the reporter moiety to the multimerized ubiquitin fusion protein. In this embodiment, enzymatic modification of the linker results in a modulation of the coupling of the reporter protein to the multimerized ubiquitin domains resulting in a change in the stability of the reporter moiety. The level of the reporter moiety in the cell can then be used as a measure of the enzymatic activity in the cell. In another embodiment the invention provides for a generalized way of coordinately regulating the cellular concentration of a plurality of target proteins. In one aspect of this method, the target proteins are operatively coupled to a ubiquitin fusion protein via a linker containing a protease cleavage site. Cleavage of the linker by a protease results in uncoupling of the target protein from the multimerized ubiquitin construct, and results in an increase in the stability and concentration of the target protein.

Methods of protein destabilization and uses thereof

Engineered fluorescent proteins, nucleic acids encoding them and methods of use are provided.

Long wavelength engineered fluorescent proteins

The present invention includes a fluorescent compound that can detect an activity, such as an enzymatic activity, and exhibits quenching. The fluorescent compound can include a fluorescent protein, such as an Aequorea-related green fluorescent protein. The fluorescent compound can include a substrate site for an enzymatic activity such as a kinase activity, a phosphatase activity, a protease activity, and a glycosylase activity. The fluorescent compound of the present invention can be used to detect such enzymatic activities in samples, such as biological samples, including cells. The present invention also includes nucleic acids that encode the fluorescent compounds of the present inventions, and cells that include such nucleic acids or fluorescent compounds.

Fluorescent protein sensors of post-translational modifications

This invention provides assays for protein kinase activity using fluorescent proteins engineered to include sequences that can be phosphorylated by protein kinases. The proteins exhibit different fluorescent properties in the non-phosphorylated and phosphorylated states.

Assays for protein kinases using fluorescent protein substrates

Engineered fluorescent proteins, nucleic acids encoding them and methods of use.

This invention provides functional engineered fluorescent proteins with varied fluorescence characteristics that can be easily distinguished from currently existing green and blue fluorescent proteins. In one aspect, the invention provides nucleic acids, expression vectors and recombinant host cells comprising nucleotide sequences encoding functional engineered fluorescent proteins comprising aromatic substitutions at position 66 and a folding mutation. In one embodiment the invention provides for fluorescent proteins containing an aromatic substitution at Thr 203.

This invention provides functional engineered fluorescent proteins with varied fluorescence characteristics that can be easily distinguished from currently existing green and blue fluorescent proteins. In one embodiment the invention provides for the three dimensional structure and atomic coordinates of an Aequorea green fluorescent protein and methods for their use. In one embodiment, this invention provides a computational method of modeling the three dimensional structure of any other fluorescent protein based on the three dimensional structure of an Aequorea green fluorescent protein.

Growing community of inventors

San Diego, CA, United States of America

Andrew B Cubitt