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Design and directed evolution of an 'Edmanase' enzyme for high-throughput peptide sequencing

Award Number: R43GM130185
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 09/13/2018
PERIOD OF PERFORMANCE END DATE: 08/31/2020

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Design and directed evolution of an 'Edmanase' enzyme for high-throughput peptide sequencing - PROJECT SUMMARY/ABSTRACT Significance: Next generation sequencing (NGS) has has transformed the field of genomics and enabled broad adoption of the technology in biological, translational, and most recently clinical research. The proteomics equivalent to NGS has not yet been realized. As such, the goal of this proposal is to develop an innovative and highly-scalable digital peptide sequencing platform to effectively scale and democratize protein analysis by providing a simple robust workflow generating massively parallel peptide sequencing information. Approach: We propose to develop a single molecule digital peptide sequencing platform, using a novel enzymatic cyclic degradation scheme, paralleling Edman degradation except using mild enzymatic steps rather than harsh chemical reagents. Our assay is comprised of several key technology elements: a population of DNA tagged single molecule peptides on beads; N-terminal amino acid (NTAA) labeling with PITC (Edman chemical agent), binding of NTAA recognition agents labeled with DNA encoding tags, writing of the DNA code to a library element attached to the peptide, and enzymatic removal of a single modified NTAA using an engineered Edmanase (the deliverable of this proposal). This assay is cycled for a dozen or more times to generate a DNA library representing peptide sequence information on millions to billions of peptides that is sequenced on a standard NGS instrument. Public Health Impact: In summary, we believe our Next-Gen peptide sequencing assay usher in the era of “next gen proteomics” effectively transforming biological, translational, and clinical research, much like NGS technology has already done in the genomics space.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2022 ( Subtotal = $0 )
2022	2018	ENCODIA, INC.	11045 ROSELLE ST STE 120	SAN DIEGO	CA	92121	SAN DIEGO	USA	Biomedical Research and Research Training	000	1	3/1/2022	NEW	$0
														Subtotal = $0

Issue Date FY: 2020 ( Subtotal = $0 )
2020	2018	ENCODIA, INC.	11045 ROSELLE ST STE 120	SAN DIEGO	CA	92121	SAN DIEGO	USA	Biomedical Research and Research Training	000	1	9/1/2020	NEW	$0
														Subtotal = $0

Issue Date FY: 2018 ( Subtotal = $222,959 )
2018	2018	ENCODIA, INC	11125 FLINTKOTE AVE STE B	SAN DIEGO	CA	92121	SAN DIEGO	USA	Biomedical Research and Research Training	000	1	9/13/2018	NEW	$222,959
														Subtotal = $222,959

Grand Total All Awards = $222,959

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Design and directed evolution of an 'Edmanase' enzyme for high-throughput peptide sequencing

Award Number: R43GM130185

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 09/13/2018

PERIOD OF PERFORMANCE END DATE: 08/31/2020

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