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Versatile Antibody for Analyzing and Purifying Proteins
EIB-2600

APPLICATIONS OF TECHNOLOGY:

ADVANTAGES:

ABSTRACT:

Dominique Loqué of the Joint BioEnergy Institute (JBEI) and Wolf Frommer of the Carnegie Institution for Science have developed a versatile antibody that can be used to localize, quantify, and purify proteins that have been expressed using Gateway® compatible vectors. The Gateway® technology is a widely used recombination cloning system that allows the efficient transfer of DNA sequences into compatible expression vectors. These vectors can then be used for in vitro transcription-translation or transformed into cells of archaea, bacteria, plants, and animals. The new antibody labels proteins over the same range of applications with much greater efficiency than conventional techniques that use tags, such as GFP.

The Gateway® technology uses recombination sites, attB1 and attB2, to transfer a DNA sequence of interest into expression vectors. All the translation products of that DNA sequence subsequently carry the peptide encoded by attB2. The JBEI antibody binds exquisitely to this peptide. Therefore, it can be used to assay or purify most translated products derived from DNA sequences engineered with attB2.

Because the epitope (5-10 amino acids) of the antibody is much smaller than most conventional tags, it is less likely to interfere with the function of the protein of interest. Moreover, since the epitope is an inherent product of the recombination system, the DNA that encodes it does not require any specific Gateway® compatible vectors. Finally, the system is applicable in high throughput experiments, such as those that quantify protein expression across different cell types transformed with the same vector.

Until now, methods for labeling proteins have required that scientists develop antibodies to their individual proteins of interest or use specific vectors to affix existing tags to these proteins. These methods can be labor intensive, relatively inefficient, and sometimes applicable with only certain cells types and proteins. The JBEI antibody overcomes these limitations.

* Gateway® recombination cloning technology is a product of Invitrogen™ by Life Technologies™.

Attention potential SBIR/STTR Program applicants (FY2013, Phase 1 Release 1): Please do not contact researchers directly with technology questions. Go here to learn how you may interact with researchers during the FOA open period.

The Joint BioEnergy Institute (JBEI, www.jbei.org) is a scientific partnership led by the Lawrence Berkeley National Laboratory and including the Sandia National Laboratories, the University of California campuses of Berkeley and Davis, the Carnegie Institution for Science and the Lawrence Livermore National Laboratory. JBEI’s primary scientific mission is to advance the development of the next generation of biofuels.

DEVELOPMENT STAGE: Proof of concept.

STATUS: Patent pending. Available for licensing or collaborative research. To view the patent application as filed, click here.

SEE THESE OTHER BERKELEY LAB TECHNOLOGIES IN THIS FIELD:

J5: Automated DNA Assembly Design Software, ECRB-2836

High Throughput Purification and Identification of Cellular Protein Complexes, JIB-2538

Optimum Protein Solubility Screen, IB-2005

REFERENCE NUMBER: EIB-2600

 

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Last updated: 10/22/2013