Research Groups
Technologies in Core Facilities
Join an RG!
Often referred to as the heart and soul of the ABRF, Research Groups (RGs) are organized by ABRF members to advance specific biotechnologies and analytical techniques for the benefit of core and research laboratories. This is done primarily by developing research studies whereby participating laboratories can gauge their ability to perform a given analytical technique(s) and, importantly, to gauge the effectiveness of that technique or methodology in real laboratory situations.
RG members typically participate for up to three years to design, conduct and report results of a specific study project. Some RG members may continue with the group for a second, three-year cycle.
RG areas of expertise cover a wide range of biotechnologies; please see the list of RG's below.
Roles within a Research Group
While many ABRF members work in the technology areas addressed by Research Groups (RGs), the success of ABRF RGs depends upon the active participation of members who contribute in various ways, including:
- Collecting samples
- Conducting tests and analysis
- Contributing supplies and services to support studies
- Preparing manuscripts for publication
- Presenting study results at ABRF and other meetings
ABRF offers online community engagement options for core personnel who are interested in following developments in these technology areas but may not be available to contribute to the efforts of an RG.
Joining an ABRF Research Group
RG areas of expertise cover a wide range of biotechnologies, and change over time as technologies change - please see the list of current RGs below. If you are interested in their research studies, have an idea for a research group in a different technology, or want to become involved in a research group, please complete this form. Your interest will be forwarded to the appropriate RG chair. Ideas for new RGs will be vetted by the Executive Board.
See a summary of current ABRF Research Group studies.
ABRF Research Groups:
Proteomics, Metabolomics, & Mass Spectrometry
Research Group Resources
- RG Handbook
- Data use guidelines to be sent with each RG study example
- Information for sponsors supporting RG studies
- Orientation presentation for new Research Group and Committee Chairs
- Project Expense Reimbursement Form
- Conflict of Interest Disclosure
Past Research Groups
Mission
The primary mission of the ABRF GlycoProtein Research Group (gPRG) is to educate the scientific community on the best techniques/strategies currently available to characterize glycoproteins.
Questions or interest in joining an ABRF research group? Contact us!
This education will be performed by offering workshops (or potentially a course) at the annual ABRF conferences so that researchers can learn the techniques currently used. The information/protocols provided during these workshops willbe complemented by having annual studies on various aspects of glycoprotein characterization that would allow individual labs to determine their competence while identifying the current “best approaches”. This research group plans to cover issues including, glycan structure determination, glycosylation site identification, and site specific glycan characterization, and will cover both N- and O-linked glycosylation. Each of these issues would be ideal for study topics. The overriding theme of this research group is to fill the current void in expertise when it comes to the characterization of the glycan portion of glycoproteins.
Past Membership
Dr William G Hendrickson (EB liaison) - Univ. of Illinois, Chicago
Dr Karen R Jonscher - University of Colorado Denver
Dr Nancy Leymarie - Center For Biomedical Mass Spectrometry, BUMS
Ron Orlando - CCRC/UGA
Studies
| 1) | Announcement for 2011 Quantitative Glycoprotein Study - View Document (86K) |
| 2) | Results for gPRG 2010 Study on Quantitative Glycoprotein Analysis - View Study Results |
Membership History
| Member Name | Organization | Details |
|---|---|---|
| Dr. Wolfgang M Egge-Jacobsen | University of Oslo, IMBV | Member: 02/09 - 09/11 |
| Mr Rodney Keck | Genentech, Inc. | Member: 01/09 - 10/10 |
| Ron Orlando | CCRC/UGA | Chair: 07/08 - 09/11 |
| Prof. Joseph Zaia | Boston University | Member: 09/08 - 09/11 |
Questions or interest in joining an ABRF research group? Contact us!
Mission
The mission of the Protein Sequencing Research Group (PSRG) is to design studies to assess techniques for determining the primary structure of protein termini that will allow participating laboratories to gauge their expectations and competence in performing these analyses.
Questions or interest in joining an ABRF research group? Contact us!
PSRG Guidelines:
The PSRG has the responsibility to regularly design and distribute samples to the ABRF membership following a reasonable time line decided mutually by the PSRG and the Executive Board of ABRF (EB). This includes sample design (EB approval required), sample preparation, test evaluation by PSRG members, sample distribution, data receipt and evaluation, oral and/or poster presentation at the ABRF or other international meeting (e.g., Protein Society or ASBMB), and finally, publication in a peer reviewed journal. The study should be designed for N – and/or C-terminal sequence analysis of proteins containing the 20 common amino acids plus (optionally) other post-translationally modified amino acids that are commonly found in proteins. Any technology that yields information about the termini of proteins may be used. The PSRG should be comprised of 6-8 members plus one EB Liaison whose purpose is to assist the research group and to facilitate communication with the EB. The PSRG should meet about once a month via a conference call to discuss the current sample and ensure continued progress. Once a year, one of the members will be chosen as chairperson in a fashion determined by the PSRG. The chair has the ultimate responsibility for ensuring the success of the RG and its studies, from initial design to the final publication. This includes communication with the EB, management office, JBT or other journal, chairs of the other protein research groups, ABRF membership, and the yearly ABRF meeting organizers. Every year, 2-3 members rotate off the RG and are replaced by new members who are nominated by the PSRG for approval by the EB. The PSRG should use this as a guideline but is not limited to it, either in order presented or type of sample. To help ensure that the PSRG meets the continuing needs of ABRF members, each sample submission form would contain a few survey questions to better gauge the kinds of samples that would be most useful to our members and to solicit suggestions for future studies.
Past Membership
David Wood, Saint Louis University, Chair
Hediye Bromage, New York University School of Medicine
Allis Chien, Stanford University
Greg Cavey, Innovative Mass Spectrometry, LLC
Robert English, Shimadzu Scientific Instruments
Brian Feild, Shimadzu Scientific Instruments
Xuemei Luo, UTMB - Galveston
Sara McGrath, U.S. Food and Drug Administration
Magnus Palmblad, Leiden University Medical Center, (EB liaison)
| 1) | PSRG 2015-2016 Study: - View Study Announcement (34K) - View Sample Insert (504K) - View Oral Presentation (2.1M) |
| 2) | PSRG 2014-2015 Study: N-terminal sequencing of standard proteins by dimethyl labeling and bottom-up mass spectrometry - View Study Announcement (34K) - View Sample Insert (359K) |
| 3) | PSRG 2013-2014 Study: N-terminal sequencing of standard proteins by N-terminal labeling and mass spectrometry - View Study Announcement (30K) - View Sample Insert (237K) |
| 4) | PSRG 2012-2013 Study : Sample preparation and terminal sequencing of a protein mixture - View Document 2012-2013 Study Announcement - View Document 2012 Participant Results Survey |
| 5) | PSRG 2011 Study: Comparing sensitivity limits for N-terminal Sequencing Technologies - View PSRG 2011 Study Announcement - View PSRG 2011 Study Sample Insert (36K) |
| 6) | PSRG 2010 Study: Comparing Edman and MS Sequencing of a Monoclonal Antibody (study closed) - View Study Announcement (94K) - View Cover Letter/Instructions (20K) - View Google Doc's data reporting form and survey |
| 7) | ESRG 2009 Study: Comparing Edman and Mass Spectrometry Techniques for N-terminal Sequencing - ESRG 2009 Study Announcement - Cover Letter/Sample Instructions (15K) - Vendor ABRF Study Participation Guidelines (18K) - Data Sheet - Excel file for both Edman and MS Data |
| 8) | ESRG 2008 Study "Investigation into homopolymeric amino acid N-terminal sequence tags and their effects on automated Edman degradation" - ESRG 2008 Study Announcement - Cover Letter/Sample Instructions - Data Sheet (41K) - Vendor ABRF Study Participation Guidelines (18K) |
| 9) | ESRG 2007 Study "Procedures used to Determine the N-Terminal Sequence of a Blocked Protein" - ESRG 2007 Study Announcement - Cover Letter/Sample Instructions - Deblocking Cycle - Data Sheet (25K) |
| 10) | ESRG 2006 Study "Edman Sequencing as a Method for Polypeptide Quantitation" - ESRG 2006 Study Announcement |
| 11) | ESRG 2005 Study "Identification of Modified Amino Acids by Edman Sequencing" - Instructions for Filling Out the Datasheet - Sample Description Including Modified Amino Acids - Data Sheet (19K) |
| 12) | ESRG 2004 Study "Modified Amino Acids in Edman Sequencing" - Sample Description Including Modified Amino Acids |
| 13) | ESRG 2004 Study Aid: Pages 10-12 from "Identification of Modified PTH-Amino Acids in Protein Sequence Analysis" 1st ed. compiled by Mark Crankshaw and Greg Grant - Chromatogram: page 10 - Legend to chromatogram: page 11 - Legend continued: page 12 |
Activities
| 1) | ESRG Newsletter Vol.1 No.1, - Report on the ESRG at ABRF 2008, Salt Lake City, Utah and a Tribute to Pehr Edman |
Electronic Posters
| PSRG 2015 - ABRF Poster and Oral Presentations: N-terminal Identification of a Standard Protein at Low Picomole Levels by Dimethyl Labeling and Bottom-up Mass Spectrometry | |
| PSRG 2014 - ABRF Poster and Oral Presentations: N-terminal Sequencing of Standard Proteins by N-terminal Labeling and Mass Spectrometry | |
| PSRG 2013 - ABRF Poster and Oral Presentations: Results of the PSRG 2013 Study Year 2: Terminal Sequencing of Standard Proteins in a Mixture |
|
| PSRG 2012 - ABRF Poster and Oral Presentations: Results of the PSRG 2012 Study: Terminal Sequencing of Standard Proteins in a Mixture, Year 1 of the 2 Year Study | |
| PSRG 2011 - ABRF Presntations: Results of the PSRG 2011 Study: SensitivityAssessment of Edman and Mass Spectrometric Terminal Sequencing of an Undisclosed Protein - PSRG 2011 Poster (4,387K) - PSRG 2011 Oral Presentation (9,213K) |
|
| PSRG 2010 Poster: Edman and Mass Spectrometric Terminal Sequencing of a Monoclonal Antibody | |
| PSRG 2010 Oral Presentation: N-Terminal Sequencing of an Antibody | |
| ESRG 2009 Poster: Comparison of Edman and Mass Spectrometry Techniques for N-terminal Sequencing | |
| ESRG 2009 Oral Presentation. "N-Terminal Sequencing Techniques" | |
| ESRG 2008 Oral Presentation. "Effects of a Homopolymeric Amino Acid Tag on Edman Degradation" | |
| ESRG 2008 Poster. "Investigation into Homopolymeric Amino Acid N-terminal Sequence Tags and their Effects on Automated Edman Degradation" | |
| ESRG 2007 Poster: "Procedures used to Determine the N-Terminal Sequence of a Blocked Protein" | |
| ESRG 2007 Oral Presentation: "Procedures used to Determine the N-Terminal Sequence of a Blocked Protein" | |
| ESRG 2006 poster: "Edman Sequencing as a Method for Polypeptide Quantitation" | |
| ESRG 2006: Oral presentation: Edman Sequencing as a Method for Polypeptide Quantitation | |
| ESRG 2005 "State of Edman Sequencing" Survey, used for both the poster and oral presentation at the ABRF meeting, Savannah, GA, February 5-8, 2005 | |
| ESRG 2005 poster "Identification of Modified Amino Acids by Edman Sequencing" | |
| ESRG 2005: Oral presentation: Modified amino acids by Edman sequencing | |
| "ABRF ESRG 2004 Study: Modified Amino Acids in Edman Sequencing"; ESRG study poster presented at the 2004 ABRF meeting February 28-March 2 in Portland, Oregon. | |
| "Modified Amino Acids in Edman Sequencing"; ESRG study results presented at the 2004 ABRF meeting February 28-March 2 in Portland, Oregon. | |
| "ABRF-ESRG 2003: Analysis of a PVDF-Bound Known Protein with a Homogeneous Amino-Terminus" ; ESRG study poster presented at the 2003 ABRF meeting February 10-13 in Denver, Colorado. | |
| Slides from tutorial, "Making the Most of Your Edman Sequence Data; A Primer on Data Calling, Analysis, Interpretation and Reporting" in pdf format, presented at the 2003 ABRF meeting February 10-13 in Denver, Colorado. | |
| Slides from the 2003 ESRG Study oral presentation given at the 2003 ABRF meeting, Feb 10-13 in Denver, Colorado. | |
| "ABRF-2002 ESRG, a Difficult Sequence: Analysis of a PVDF-Bound Known Protein With a Heterogeneous Amino-Terminus"; poster presented at the 2002 ABRF meeting March 10-12 in Austin, Texas. | |
| Slides from ESRG 2002 Study oral presentation given at 2002 ABRF meeting March 10-12 in Austin, Texas | |
| The first half of the tutorial session "N-terminal Edman sequencing sample preparation" slides in pdf format. | |
| Second half of the tutorial session "N-terminal Edman sequencing sample preparation" slides in pdf format. | |
| Here is the method briefs handout from the tutorial session. Please use these methods and references only as guides for you to develop similar methods more fully in your own labs, no liability to ABRF for their use. |
Membership History
| Member Name | Organization | Details |
|---|---|---|
| Daniel Brune | Arizona State Univ | Chair: 02/05 - 02/06 Member: 02/06 - 04/07 |
| Scott Buckel | Amgen, Inc. | Member: 01/01 - 03/03 |
| Dr. Allis Chien | Stanford University | EB Liaison: 03/17 - 09/18 |
| Richard G. Cook | Baylor Col of Med | Member: 01/01 - 03/04 |
| J. M. Crawford | Yale Univ. Keck Biotech Res. Lab. | Member: 01/01 - 03/04 |
| Dr Nancy D Denslow | Univ of Florida-Gainesville | Chair: 03/04 - 02/05 Member: 03/02 - 03/04 EB liaison: 02/05 - 02/09 |
| David R. Dupont | Applied Biosystems | Member: 01/01 - 03/02 |
| Dr Robert English | Univ. of Texas Medical Branch | Chair_(co-chair): 04/12 - 04/13 Member: 05/11 - 04/12 |
| Dr Hediye Erdjument-Bromage | Sloan Kettering Inst | Member: 04/14 - 04/15 |
| Joseph Fernandez | Rockefeller Univ | Chair: 01/01 - 03/02 Member: 03/02 - 03/03 |
| Mr. Mark K. Garfield | NIAID/NIH | Member: 05/12 - 04/15 |
| Brian Hampton | Universtiy of Maryland School of Medicine | Member: 02/05 - 04/07 Member: 02/08 - 02/09 Ad hoc: 02/09 - 04/10 Co-Chair: 04/07 - 02/08 |
| Dr William G Hendrickson | Univ. of Illinois, Chicago | EB Liason: 03/13 - 04/14 |
| Peter Hunziker | University of Zurich | Chair: 02/08 - 02/09 MemberBecome Ad Hoc: 02/09 - 04/10 Member: 03/06 - 02/08 Ad hoc: 04/10 - 04/11 |
| Pegah R Jalili | Sigma-Aldrich | Member: 06/11 - 04/14 |
| Viswanatham Katta | Genentech | Member: 06/10 - 03/13 |
| Ryuji Kobayashi | UT MD Anderson Cancer Center | Member: 03/03 - 03/06 |
| William S. Lane | Harvard University | EB Liason: 10/03 - 02/05 |
| Joseph W. Leone | Pfizer | Chair: 02/06 - 04/07 Member: 03/04 - 02/06 Member chair-emeritus: 04/07 - 02/08 |
| Joseph F. Leykam | Michigan State University | Member: 04/07 - 08/07 |
| Dr. Klaus D. Linse | Biosynthesis Inc. | Member: 02/05 - 02/08 |
| Benjamin J Madden | Mayo Clinic | Chair: 02/03 - 03/04 Member: 03/04 - 02/05 Member: 01/01 - 02/03 |
| Kwasi G Mawuenyega | Washington University School of Medicine | Member: 05/09 - 04/12 |
| Dr. Sara C. McGrath | US FDA | Chair_(co-chair): 04/13 - 04/15 Member: 05/12 - 03/13 |
| Dr. Ejvind Mortz | Alphalyse | : 03/13 - 04/14 |
| John M Neveu | Harvard University | Chair: 04/02 - 01/03 Member: 01/01 - 03/02 Member: 02/03 - 02/05 |
| Dr. Ronald L. Niece | Research Resources & Technologies | Member: 03/88 - 03/91 |
| Dr. Brett S Phinney | Proteomics Core UC Davis Genome Center | EB Liason: 04/14 - 04/15 |
| Dr Jan Pohl | Emory Univ Sch of Med | Member: 03/04 - 04/07 |
| Dr. Henriette A. Remmer | University of Michigan | Member: 06/10 - 04/11 Ad hoc: 03/13 - 04/15 co-chair: 04/11 - 04/12 Co-chair: 04/12 - 03/13 |
| Wendy Sandoval | Genentech, Inc. | Member: 04/10 - 04/11 Member changing to cochair: 04/07 - 02/09 Co-chair: 02/09 - 04/10 |
| Jack Simpson | US Pharmacopeia | EB liaison: 02/09 - 03/13 |
| John S. Smith | Univ of Texas Med Branch | Member: 04/10 - 04/11 Member changing to cochair: 09/07 - 02/09 Co-chair: 02/09 - 04/10 |
| Laurey Steinke | University of Nebraska Medical Center | EB liason: 01/01 - 09/03 |
| Dr. Detlev Suckau | Bruker Daltonik GmbH | MemberBruker Daltonics: 05/10 - 03/13 |
| Richard S Thoma | Monsanto | Member: 02/08 - 02/09 Member: 02/05 - 04/07 Ad hoc: 02/09 - 04/10 Co-Chair: 04/07 - 02/08 |
| James Walters | Sigma-Aldrich | Chair: 04/10 - 04/11 Member: 04/09 - 04/10 Ad hoc: 04/12 - 03/13 co-chair: 04/11 - 04/12 |
| Dr. Frances Weis-Garcia | Memorial Sloan Kettering Cancer Center | EB Liaison: 02/16 - 03/17 |
| Ken R. Williams | Yale Univ, Keck Lab | Member: 03/88 - 03/90 |
| Dr. Bosong Xiang | Monsanto Company | Member: 04/09 - 04/12 |
Publications
-
A Synthetic Peptide for Evaluating Protein Sequencing Capabilities: Design of ABRF-93SEQ and Results. In (J.W. Crabb, ed.) Techniques in Protein Chemistry V, Academic Press, San Diego, pp133-141 (1994)Rush, J., Andrews, P. C., Crimmins, D. L., Gambee, J. E., Grant, G. A., Mische, S. M. and Speicher, D. W.
-
ABRF ESRG 2005 Study: identification of seven modified amino acids by Edman sequencing. J Biomol Tech, 2006. 17(5): p. 308-326.Brune, D., N.D. Denslow, R. Kobayashi, W.S. Lane, J.W. Leone, B.J. Madden, J.M. Neveu, and J. Pohl
-
ABRF-2002 ESRG, A Difficult Sequence: Analysis of a PVDF-Bound KNown Protein With a Heterogeneous Amino-Terminus Journal of Biomolecular Techniques 13:246-257 (2002).Buckel, S.D., Cook,R.G., Crawford, J.M., Dupont, D.R., Madden,B.J., Neveu,J.M., Steinke, L., Fernandez, J
-
ABRF-2003 ESRG: Analysis of a PVDF-Bound KNown Protein With a Heterogeneous Amino-Terminus Journal of Biomolecular Techniques 14:278-288 (2003).Buckel, S.D., Cook,R.G., Crawford,M.J., Denslow, N., Madden,B.J., Steinke, L., Fernandez, J., Neveu,J.M
-
ABRF-97SEQ: Sequencing Results of a Low-Level Sample. J Biomol Tech, 10:26-32 (1999)Stone, K., Fernandez, J., Admon, A., Henzel, W., Lane, W., Rohde, M., and Steinke, L.
-
ABRF-98SEQ: Evaluation of peptide sequencing at high sensitivity J. Biomol. Tech., Jun 2000; 11: 92 - 99.WJ Henzel, A Admon, SA Carr, G Davis, K De Jongh, W Lane, M Rohde, and L Steinke
-
Amino Acid Analysis and Sequencing - What Is State-of-the-Art?Niece, R.L., L.H. Ericsson, A.V. Fowler, A.J. Smith, D.W. Speicher, J.W. Crabb, and K.R. Williams.
"Methods in Protein Sequence Analysis" (H. Jörnvall, J.-O. Höög, and A.-M. Gustavsson, Eds.), Birkhäuser Verlag, Basel, 133-141, 1991
-
Assignment of Cysteine and Tryptophan Residues during Protein Sequencing: Results of ABRF-94SEQ. In (J. W. Crabb, ed.) Techniques in Protein Chemistry VI, Academic Press, San Diego, pp 209-217 (1995)Gambee, J. E., Andrews, P. C., Grant, G. A., Merrill, B., Mische, S. M. and Rush, J.
-
Design and Analysis of ABRF-95SEQ, a Recombinant Protein with Sequence Heterogeneity. In (D. Marshak, ed.) Techniques in Protein Chemistry VII, Academic Press, San Diego, pp 347-358 (1996)DeJongh, K. S., Fernandez, J., Gambee, J. E., Grant, G. A., Merrill, B., Stone, K. L. and Rush, J.
-
Design, Characterization and Results of ABRF-89SEQ: A Test Sample for Evaluating Protein Sequencer Performance in Protein Microchemistry Core Facilities. In (T. E. Hugli, ed.) Current Research in Protein Chemistry, Academic Press, pp 159-166 (1989)Speicher, D. W., Grant, G. A., Niece, R. L., Blacher, R. W., Fowler A. V. and Williams, K. R.
-
Edman Sequencing Research Group 2004 Study: Modified Amino Acids in Edman Sequencing, J. Biomol. Tech., Sep 2005; 16: 272 - 284.D. Brune, J.M. Crawford, R.G. Cook, N.D. Denslow, R. Kobayashi, B.J. Madden, J. M. Neveu, and L. Steinke
-
Evaluation of ABRF-96SEQ: A Sequence Assignment Exercise. In (D. R. Marshak, ed.) Techniques in Protein Chemistry VIII, Academic Press, San Diego, pp 69-78 (1997)Fernandez, J., Admon, A., DeJongh, K., Grant, G., Henzel, W., Lane, W. S., Stone, K. L. and Merrill, B.
-
Evaluation of Protein Sequencing Core Facilities: Design, Characterization, and Results form a Test Sample (ABRF-91SEQ). In (R. H. Angeletti, ed.) Techniques in Protein Chemistry III, Academic Press, San Diego, pp 35-35 (1992)Crimmins, D. L., Grant, G. A., Mende-Muller, L. M., Niece, R. L., Slaughter, C., Speicher, D. W. and Yuksel, K. U.
-
Instrumentation Used in Protein and Nucleic Acid Resource Facilities: A Survey of UsersBeach, C.M. G.M. Hathaway, T.K. Hayes, A.J. Smith, and R.L. Niece.
ABRF News, Vol. 2.1 Supplement: 1-7, 1991
-
Protein Sequencing from Polyvinylidene Difluoride Membranes: Design and Characterization of a Test Sample (ABRF-90SEQ) and Evaluation of Results. In (J. J. Villafranca, ed.) Techniques in Protein Chemistry II, Academic Press, San Diego, pp 151-162 (1991)Yuksel, K. U., Grant, G. A., Mende-Muller, L. M., Niece, R. L., Williams, K. R. and Speicher, D. W.
-
Protein Sequencing of Post-translationally Modified Peptides and Proteins: Design, Characterization and Results of ABRF-92SEQ. In (R. H. Angeletti, ed.) Techniques in Protein Chemistry IV, Academic Press, San Diego, pp 453-461 (1993)Mische, S. M., Yuksel, K. U., Mende-Muller, L. M., Matsudaira, P., Crimmins, D. L. and Andrews, P. C.
-
in "Techniques in Protein Chemistry" (Hugli, T., Ed.), Academic Press, San Diego, 89-110, 1989
2000 ABRF Peptide Sequencing by Mass Spectrometry Quiz
Introduction
Prior to its demise, the ABRF Mass Spectrometry Committee prepared four "tryptic" peptides whose sequences did not match anything in the public nucleotide or protein sequence databases. For two of these peptides, data was acquired using an ion trap mass spectrometer, and for the other two a quadrupole / time-of-flight hybrid mass spectrometer (Qtof) was used. Using these data, a MS/MS peptide sequencing quiz was prepared for the ABRF membership, where the aim was four-fold. First, this was a sort of late final exam for those who took the 1999 ABRF course taught by Prof. Don Hunt. Second, the results might indicate the need for similar additional ABRF courses. Third, the anonymous results allowed individuals to assess their own level of proficiency at this task. Fourth, it allowed a comparison of the relative merits of two types of mass spectrometers most commonly used for peptide sequencing.
This study is closed, and the results and answers can be found here. I suppose one could still take the quiz, but you'll have to grade yourself.
The Quiz
Check out the sequencing tutorial, if you need instructions on sequencing tryptic peptides using low energy CID data.
Ion trap data
Data for two peptides was acquired, where the first one is the easiest. To aid in the verification of your proposed sequences, MS3 spectra were acquired for both peptides. The following are all standard gif files:
- ABRF MS Sequencing Problem #1.
- MS3 for ABRF MS Sequencing Problem #1
- ABRF MS Sequencing Problem #2
- MS3 for ABRF MS Sequencing Problem #2
Qtof data
The next two problems were obtained from a quadrupole / time-of-flight hybrid mass spectrometer (Qtof), where the third peptide is easier than the fourth. The fourth peptide was also subjected to trypsin in the presence of a 1:1 mixture of 16O and 18O water, which labeled the C-terminus with a mixture of the two stable isotopes. The precursor resolution was set wide enough to pass both the 16O and 18O labeled peptide, such that C-terminal fragment ions in the MS/MS spectrum exhibited two mass unit doublets. There are two spectra shown, where the raw data is on top, and the spectrum beneath it is the same data subjected to a y-ion filter.
- ABRF MS Sequencing Problem #3 (16O only)
- ABRF MS Sequencing Problem #4 (16O only)
- ABRF MS Sequencing Problem #4 (C-terminus labeled with 16O and 18O). To view this you will need a copy of Acrobat Reader, which can be obtained from the Adobe Acrobat web site.
Synthetic Peptide for Evaluating Protein Sequencer and Amino Acid Analyzer Performance in Core Facilities: Design and ResultsNiece, R.L., K.R. Williams, C.L. Wadsworth, J. Elliott, K.L. Stone, W.J. McMurray, A. Fowler, D. Atherton, R. Kutny, and A.J. Smith. A.
Questions or interest in joining an ABRF research group? Contact us!
September 2021 – ABRF NGS Study featured in Nature Biotechnology
See related story in GenomeWeb.
This online ABRF seminar, produced in collaboration with GenomeWeb, details the results of the first phase of an ongoing study being conducted by ABRF Research Groups that aims to evaluate the performance of six next-generation sequencing platforms: Illumina HiSeq 2000/2500, Illumina MiSeq, Roche 454 GS FLX, Life Technologies Ion Torrent PGM, Life Technologies Ion Torrent Proton, and Pacific Biosciences PacBio RS.
The first phase of the study focused on transcriptome analysis and involved more than 20 core facility laboratories who used standard reference samples from the Microarray Quality Control Consortium to perform replicate RNA‐seq experiments on these platforms. A paper describing the study was recently published in Nature Biotechnology.
This webinar will discuss the findings from this phase, which showed high intra‐platform consistency and inter‐platform concordance for expression measures, but also demonstrated highly variable rates of efficiency and costs for splice isoform detection between platforms. The speakers will also describe a comparison of alternative aligners for each platform, which showed that algorithm choice affects mapping rates and transcript coverage more than gene quantification.
The speakers will also provide an overview of the next phase of the study, which is focused on DNA reference standards, and will discuss the study’s goal of establishing a community resource that will allow users of sequencing technologies to easily compare performance data as instruments and protocols change.
Panelists:
- George Grills, Cornell University, ABRF Executive Board
- Scott Tighe, University of Vermont, ABRF Nucleic Acid Research Group
- Christopher Mason, Weill Cornell Medical College, ABRF Genomics Bioinformatics Research Group
- Don Baldwin, Pathonomics LLC, ABRF Genomics Research Group
- Marc Salit, National Institute of Standards and Technology
Read the manuscript here!
Multi-platform assessment of transcriptome profiling using RNA-seq in the ABRF next-generation sequencing study.
Sheng Li, Scott W Tighe, Charles M Nicolet, Deborah Grove, Shawn Levy, William Farmerie, Agnes Viale, Chris Wright, Peter A Schweitzer, Yuan Gao, Dewey Kim, Joe Boland, Belynda Hicks, Ryan Kim, Sagar Chhangawala, Nadereh Jafari, Nalini Raghavachari, Jorge Gandara, Natàlia Garcia-Reyero, Cynthia Hendrickson, David Roberson, Jeffrey A Rosenfeld, Todd Smith, Jason G Underwood, May Wang, Paul Zumbo, Don A Baldwin, George S Grills & Christopher E Mason
Mission
Many of the Members on the MMRG have been involved in microbiome analysis for over 20 years and have a strong appreciation for the field. They are dedicated and understand the needs of this new and exciting field that continues to grow at an exponential rate.
Mission
The Proteomics Research Group (PRG) is a volunteer scientific organization dedicated to sharing knowledge about the analysis of proteins. The PRG aims to assist protein scientists and resource facilities to achieve their highest potential by sponsoring annual research studies that examine current techniques and capabilities. Through the promotion of broad participation and scientific excellence, the PRG aims to raise awareness, knowledge and education about modern methods of protein analysis.
Mission:
