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Fast, flexible, and economical system for affinity-tag protein purification and tag removal

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: 1R43GM126676-01
Agency Tracking Number: R43GM126676
Amount: $224,994.00
Phase: Phase I
Program: SBIR
Solicitation Topic Code: 300
Solicitation Number: PA16-302
Timeline
Solicitation Year: 2016
Award Year: 2018
Award Start Date (Proposal Award Date): 2018-04-05
Award End Date (Contract End Date): 2019-04-04
Small Business Information
11305 DUNLEITH PLACE
Gaithersburg, MD 20878-2566
United States
DUNS: 193771347
HUBZone Owned: No
Woman Owned: Yes
Socially and Economically Disadvantaged: No
Principal Investigator
 PHILIP BRYAN
 (240) 314-6220
 pbryan@umd.edu
Business Contact
 BONNIE BRYAN
Phone: (240) 252-9749
Email: bbryan@potomac-affinity-protein.com
Research Institution
N/A
Abstract

Project SummaryOur long term objective is to commercialize a fastsimpleflexibleand economical
system for affinity tag protein purification and tag removalThis technology was developed as a result of
NIH funded studies on the foldingstability and enzymology of the Bacillus protease subtilisinThe
transformative purification technology combines three componentsa regulatedhighly specific
proteasePsubthat is constitutively inactivea small moleculeimidazolethat activates the Psuba high affinity inhibitorProImmobilized Psubin the off statecan capture a Pro fusion protein from a
cellular extractallowing contaminants to be washed awaySubsequent addition of imidazole to the
immobilized complex releases puretag free target protein and leaves the Pro tag tightly bound to PsubRecombinant proteins are frequently fused with other proteins or peptides to facilitate expression and
purificationThe tags enable target protein binding in affinity purificationbut ultimately must be
processed by a site specific proteaseTag removalhoweveris frequently expensiveinefficientand
sometimes problematicThe technical innovation of our system is the integration of seamless tag
removal into the purification processThis provides simplicityefficiencyand robustness that is not
available in any other systemImproved understanding of protein engineering principles has resulted in
highly efficient Psubs and Pro affinity tagsWe will develop and test purification materials and methods
around imidazole triggered chemistry and improved affinity tagsThe Specific Aims areproduce
imidazole triggered Psubs at low costclone a range of test proteins into expression vectors with the
new tagsMeasure soluble expression for the Pro tagged test proteinsDevelop and test
purification methods using Psubs and test proteins chosen to challenge purification methodsRefine
Psubs if needed by screening phage librariesThe system eventually should be suitable for both highthroughputpurification of proteins on the laboratory scaleas well as process scale purification of
pharmaceutical proteinsBetter purification technology should advance in all areas of biology Project NarrativeChallenges in protein purification limit progress in biology and
medicineThe purification technology being developed in this project should benefit
anyone purifying proteinsbut particularly improve study of many challenging and
medically important eukaryotic proteinsThe technology should eventually benefit largescale purification of pharmaceutical proteins

* Information listed above is at the time of submission. *

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