February 2003
Creating a CMC-IGERT Team
(Forms A & B)
This document can be found at:
http://macro.lsu.edu/igert/ActiveServerPages/CurrentStudents.asp
It should open as a Word document, ready for you to modify.
Making a new IGERT team involves three steps:
1. Complete “Form A”, which is a written proposal in the particular format shown by
examples in the Appendix of this document. These examples are taken from
Section D of the original proposal. For your convenience, an empty template
follows on page 2 of this document. Just cut & paste it into Microsoft Word
(please!) and fill in the blanks after talking out your project as a team.
2. Enter data for students, faculty and off-campus personal data into our web form:
http://macro.lsu.edu/igert/ActiveServerPages/DataSheet.asp
3. Complete “Form B” after your team is accepted. Each member must sign,
indicating full knowledge of the IGERT guidelines, purposes and policies.
“Form B” also contains a more detailed Apprenticeship plan and timeline than
“Form A.”
You may have completed the first step already!

If you were one of the original participants, and if you still propose to do the
project that appeared in Section D of the proposal without significant
modifications, you already completed Step 1. Just go to the website and fill out
the "New Team" data form.

If you were one of the original participants whose project appeared in capsule
form in Section E, we have to decide whether or not you have completed Step 1
already. You may have to fill out the NEW IGERT TEAM FORM. Call Russo,
Dooley or Bricker. After the form has been filled out, you go to the website for
data entry.

If you were not a participant on the grant at all, or are proposing an entirely
different project, you definitely must fill out the NEW IGERT TEAM FORM and
then go to the website.
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STEP 1 = Form A (Proposal to create a new IGERT team)
You can cut & paste the text below (starting with D1. Title Title Title) into Microsoft
Word and "fill in the blanks". Send the completed Form A to the IGERT Coordinator,
Ms. Florence Schmitt (fschmi1@lsu.edu) Do NOT send the form to any of the directors.
Please check the examples provided in the Appendix for content and style. Your
proposal can be somewhat shorter, especially if it is very risky new research, but it must
contain each the following sections.
D1. Title Title Title Title
This project will:
Primary Faculty co-Advisors (at least 2!)
Name1, Department (Research Specialty)
Name2, Department (Research Specialty)
Name3, Department (Research Specialty)
Off-campus Participant: Name (University, Company or Institute)
Technical Proposal: Think in terms of text and pictures that will look good when
mounted on the website (don't worry, we will remove proprietary stuff if you want).
The purpose is to clearly explain your project to site visitors, including potential
students who may wish to add in to your team.
Number of IGERT apprentices to be recruited and probable home departments: Here, just
state the number and their departments. We'll get to names later!
Consistency with the Macromolecular Education, Research & Training theme: Why is this
project consistent with project goals? We can discuss that if you aren't clear about it.
How does the project form a vector cross-product of existing research themes by the
participants?
Existing research directions. State what each faculty member is now doing and give some idea of
the Federal, state and industrial support for that endeavor and/or plans to obtain support.
New research direction. State what each faculty member will do that he or she could not do
without the team approach.
How do students benefit from the team-oriented research, beyond what would be available
to them from either advisor separately? See examples in Appendix.
Briefly describe the support level available to each individual faculty or off-campus
participant (i.e., without IGERT) See examples in Appendix. For very good reasons, the
IGERT grant cannot be the only support mechanism for its professors. In short, if you need it,
you cannot have it. If you need it to take your active research group in a wholly new direction,
that is another matter!
Interdisciplinary strengths of the team project: See examples in Appendix.
Commitment of faculty & off-campus participants to work side-by-side with apprentices:
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Say what each professor plans to commit during the apprenticeship period. IGERT intends to
put you (back) in the lab! We know that you have to break to make phonecalls and answer email, but the objective is really working with the apprentice for a very significant time. The
amount of time should be guided by how long it normally takes you to find money to fully
support a student for 5 years. For example, if you spend about 2 months preparing a standard
NSF grant for a postdoc and one student, I would suggest a time commitment to the apprentice
of 3 weeks to one month. A format like the following is helpful to the committee that approves
teams: Professor XXX, a full professor within the Department of XXXX, promises blah-blahblah. Professor YYY, an assistant professor in the Department of XXXX blah-blah-blah.
References: (ACS journal with titles format, as below--not too many are required!)
Szydlowski, J.; Rebelo, L.P.; Wilczura, H.; Dadmun, M.; Melnichenko, Y.; Wignall, G.D.; Van Hook,
W.A.: "Comparison of SANS and DLS hydrodynamic correlation lengths for a
polystyrene/methylcyclohexane solution in the vicinity of temperature or pressure induced critical
demixing," Physica B. Condensed Matter, 1998, 241/243, 1035-1037
Yeo, S. D.; Debenedetti, P. G.; Radosz, M.; Schmidt, H-W. "Supercritical Anti-Solvent (SAS) Process for
Substituted Para-Linked Aromatic Polyamides: Phase Equilibrium and Morphology Study"
Macromolecules 1993, 26, 6207 and 1995, 28, 1316.
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STEP 2: Visit the website
http://macro.lsu.edu/igert/ActiveServerPages/DataSheet.asp
Please go to the IGERT website and fill out the "New Team" Forms. You will need:
For each nominated apprentice:
 Full name
 Permanent home (i.e., parents' home) address & phone
 Home in Baton Rouge (address, e-mail and phone)
 Department
 Undergraduate institution
 Undergraduate degree and major
 Undergraduate GPA
 GRE scores (verbal, quantitative and analytical)
 GPA at LSU (if applicable; use mid-term grades if first semester)
 Current level of support (TA, RA, fellowships, scholarships, sign-on bonuses,
etc.)
 Be prepared to sign that you have fully read and understand the IGERT guidelines
and requirements (extra courses, seminars, retreats, etc.). You should revisit the
website and/or see Russo, Dooley or Bricker if you have questions.
 Hobbies
 Birth date
 Apprentice project title, faculty participant and dates
For each participating faculty member (website under development)
 Name & Department.
 Academic Rank
 E-mail & Phone
 Funding directly under your control.
 Funding partly under your control.
 Birth date
For each off-campus participant (website under development)
 Name
 Institution
 Academic or Corporate Rank
 E-mail & Phone
 Type of participation in IGERT
o Committee only
o Internship in my lab (supported by my company or university)
o Internship in my lab (supported by IGERT)
o Other (specify)
 Birth date
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Step 3 = Form B (return to Florence Schmitt)
Apprenticeship Detail
(say what you plan to do and when; keep it short but be prepared to live up to your
commitment! Here is one example; you can just replace the text.)
Student name: Nadia Edwin
For approximately 3 weeks during May, 2001, Ms. Edwin will work with Prof. Russo and
selected members of his group on diffusion ordered NMR spectroscopy (DOSY). It is
anticipated that Prof. Russo can spend 5-6 hours per day during this period on task (in the
laboratory, working directly with the student). They will measure diffusion coefficients
of a variety of small and large polymers by DOSY and compare to dynamic light
scattering (DLS). In selected cases, they will compare to Fluorescence Photobleaching
Recovery (FPR) and analytical ultracentrifugation (AUC). The purpose of the project is
to establish, for the first time at LSU, the relative performance and limitations of these
methods. Adaptation of Bruker DOSY data output to the Graphical User Interface (GUI)
Laplace inversion software within the Russo group is another objective, to increase the
convenience of using DOSY. The apprenticeship also exposes Ms. Edwin to Drs. Frank
Zhou, Dr. Dale Treleavan, and Dr. Rafael Cueto, all members of the Chemistry
Department or Basic Sciences Technical Staff. Jason Campbell will "shadow" this
research project, as his interests include DLS and DOSY. Additionally, Ms. Edwin will
"shadow" the Small Angle X-ray Scattering construction project, which involves
Professors Thomas, Russo and Negulescu, off-campus participant Prof. Greg Beaucage
and trainees Thomas Morgan, Mariah McMasters, and Jason Campbell.
Plans to seek additional support once some preliminary data have been obtained.
(specify which agency or agencies are most likely to support the work in the long term.
IGERT is to act as a catalyst for new research. Students who can be picked up on
additional, new support (old support for old projects does not apply) can remain in the
IGERT program…and will be rewarded for helping find new support. It should fit into the
space below after these bold instructions are erased).
Contingency plans.
(What will the team do if one of its participants becomes unable to complete the research?)
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Commitment
We agree to perform the apprenticeship project (side-by-side research) as described
above. We understand that the apprenticeship must culminate with the production of a
“minithesis” (examples on request) to be co-authored by the faculty and students.
Further, we understand that CMC-IGERT students and faculty participants will
participate frequently in Macromolecular seminars and assist occasionally with recruiting
and community outreach. Faculty advisors agree to help occasionally lead the teamtaught courses in the Macro core curriculum (401X courses). Students agree to enroll in
these courses plus one approved (by request) elective. All parties agree to use laboratory
notebooks provided by the CMC-IGERT program, making these (or copies) available to
confidential analysis by the IGERT evaluation team. All parties further agree to
complete external evaluation forms provided by NSF as part of its evaluation of this and
other IGERT sites. Students agree to complete other provisions of the program, such as
the STSC and PCO courses, Community Service, data defense, etc. It is understood that
IGERT Fellowship is temporary. Students and teams are reviewed each semester and a
decision is made. Our team understands that students should NOT expect 5 solid years of
stipend support; however, those who remain active in the program can often take
advantage of its other features (minigrants, finishing school, etc.) after stipend support
has ended. Other provisions of the program are well-understood from inspection of the
website, http://macro.lsu.edu/igert . In particular, it is understood that the entire project is
one educational experiment where the benefactor (NSF) is concerned. We agree to treat
it with the respect that any scientific experiment requires.
Signed (all team members must sign, except off-campus participants who are not
responsible for the side-by-side research project).
_____________________________________________________________________
printed name, signature and date
_____________________________________________________________________
printed name, signature and date
_____________________________________________________________________
printed name, signature and date
_____________________________________________________________________
printed name, signature and date
_____________________________________________________________________
printed name, signature and date
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Appendix.
Two Projects Selected from Sec. D of the Original IGERT Proposal
Consider these two examples as you design your team. Also, the lead-in text (under
“Major Research Efforts”) gives more clues about what makes a good project.
D. Major Research Efforts
A few projects will illustrate the broad base of activities that can share the interdisciplinary
educational model, administrative practice and core curriculum to be developed and tested.
Faculty teams submitted the following research descriptions in response to guidelines circulated
electronically by the co-directors. This was done to illustrate the actual process by which IGERT
participants will launch their teams. To hasten recruiting if the proposal is funded, participants
were urged to write at least a portion of their proposal in a style that would appeal to prospective
graduate students who may know little about macromolecules. We received 9 projects; the
scientific part of each has been placed on the CMC IGERT website to hasten recruiting if the
award is made. The 5 projects described in detail below reflect the breadth and balance of the
program, but are not otherwise better than those in Sec. E. All are listed on the website
(http://russo.chem.lsu.edu/igertweb). If the proposal were funded, these team leaders would be
authorized by the co-directors to populate their project with IGERT students. This signifies that
the project is appropriate for integrated macromolecular education, research and training. It also
signifies that the project directors are well-funded, active investigators who have identified a
research vector cross product--i.e., a new direction that can only be reached through teamwork.
Authorization to recruit also means that the project leaders promised to participate in the side-byside research project with their apprentice and comply with the reporting requirements of the
CMC experiment. They have located off-campus participants, or agreed to do so. If no active
researcher has been located off-site, the project leaders must at least agree to identify an offcampus committee member who will travel to LSU to serve on the Ph.D. qualifying and final
defense exams. For the project to receive actual money, the faculty partners must successfully
recruit two or more qualified students and specify their roles. Also, the off-campus participants
must be positively identified.
The value added by IGERT is the essential, catalytic support for student teams to
be shared by two or more faculty, the requirement that the team create a new
research direction, and generous seed resources to produce the data required for
sustaining support.
D1. New Technologies and Methodologies for Protein Analysis
This project will: develop new micro-machined devices for the separation of proteins and
couple these devices to high performance mass spectrometers for protein identification.
Primary Faculty co-Advisors:
Terry Bricker, Biological Sciences Department (Protein Chemistry)
Pat Limbach, Chemistry Department (Mass Spectrometry)
Steve Soper, Chemistry Department (Micro-instrumentation/Chemical Separations)
Off-campus Participant: Jon Amster (University of Georgia)
Technical Proposal: "Biochemistry is a technique-driven science. One deals today with those
problems that today's technology gives one a chance of solving, not necessarily with those
problems one would like most to solve."(1) The goal of this project is to develop new analytical
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instrumentation and protein analysis methodologies. The scientific advances which will result
from this project revolve around the development of new micro-instruments for separation and
purification of proteins and the interfacing of these devices to high performance mass
spectrometers. These micro-instruments are ideally suited for the separation and purification of
small amounts (one-billionth of a gram or less) of biomaterials and will result in a dramatic
improvement in analysis throughput, thereby allowing for a number of informative studies to be
performed to rapidly elucidate the function of various proteins. The technical and procedural
developments from this project will result in a new paradigm for biomolecule analysis.
Protein Characterization. Elucidation of the functional properties and structural
organization of membrane protein complexes is one of the central objectives of current
biochemical investigation. Biological membranes are involved in virtually every aspect of
cellular organization and activity. One of the most intriguing aspects of membranes is their role
in energy transfer in photosynthetic organisms. Light energy, which is the product of a most
violent physical process, fusion, is transformed into biological energy equivalents utilized by the
photosynthetic cell. The photosynthetic process provides both the carbohydrate which lies at the
base of virtually all food chains and, as a byproduct, all of the atmospheric oxygen. Recently,
much effort has been directed towards understanding the structure, function, and assembly of the
membrane protein complexes involved in the photosynthetic light reactions. Despite much
progress, the mechanisms involved in protein processing, membrane insertion, cofactor assembly,
and regulation of photosynthetic electron transport remain poorly understood.
In this project, we will examine the functional proteomics of the thylakoid lumen from both
higher plants and cyanobacteria. This subcellular compartment is very poorly understood and
until recently had only been cursorily examined. Preliminary results from the Bricker laboratory
and others indicate that, while more than one hundred protein components may be present in the
lumenal compartment, the functions of less than fifteen of these have been identified. We
propose to isolate and identify the proteins located in the thylakoid lumen in both higher plant
(Arabadopsis) and cyanobacterial (Synechocystis 6803) systems. These model systems are
particularly appropriate for these studies as the genome of Synechocystis 6803 has been
sequenced and that of Arabadopsis will be completed within the next two years (about 40% of
this genome is currently known). We will then systematically delete the genes encoding these
proteins in the cyanobacterial system. These experiments will allow us to form working
hypotheses as to the function of these lumenal components, which will then be tested by rigorous
molecular and biochemical approaches.
Initially, the lumenal proteins will be isolated from higher plant and cyanobacterial thylakoid
membranes. The Bricker laboratory has already developed efficient procedures for the isolation
of these components from higher plants (2) and is currently extending these techniques to
cyanobacterial thylakoid membranes. The proteins of these lumenal preparations will then be
identified as described below which will permit the determination of the genes encoding the
individual protein spots. Once the genes encoding the lumenal proteins have been identified,
insertional and/or deletion mutagenesis will be performed in Synechocystis 6803. These
techniques have already been implemented in the Bricker laboratory. Mutant screening will
allow a preliminary assessment as to the functional lesion induced by the mutagenesis and will
provide a first glimpse as to the function of these lumenal proteins.
Micro-instrumentation and Mass Spectrometry While the specific target of this research
project will be to characterize lumenal proteins, another major component will be to fabricate the
appropriate micro-instrumentation which will allow for the purification of protein mixtures, such
as those provided by the thyalkoid lumen, prior to their identification and structural
characterization by mass spectrometry. The lumenal proteins, which will be isolated in the
Bricker lab as discussed above, provide an excellent model for us to test the performance of our
micro-machined devices and to train and educate our students in the areas of protein chemistry,
micro-instrumentation, and structural analysis. As only approximately 100 lumenal proteins will
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be present, the level of complexity compared to that of another model system, such as Eschericia
coli, is reduced, thereby allowing us to examine an important and relevant biological system
without prematurely attempting a system that is too complex.
The fabrication of micro-instrumentation using photolithographic techniques has permeated
into the molecular biology and chemistry arenas and has already had a profound impact on
instrument development and assay methodologies. The principal advantages associated with
these devices include: (a) the instrument footprint is small; (b) the micro-fabricated device can be
multiplexed easily so that several analyses can be completed simultaneously; (c) complex patterns
of channels for fluid manipulation and mixing can be fabricated; (d) the potential for integrating
various components onto the device is high; and (e) the sample requirements are small.
Sample preparation, which will be a key step in our protein analysis methodology, has been
neglected to a great extent by a number of research groups developing microfabricated devices.
The difficulty in developing miniaturized devices for sample preparation is the low volume
requirement associated with these devices, creating potential difficulties in transferring the
sample from one device to another. That difficulty is somewhat reduced in our application, due
to the low sample amounts of proteins to be characterized. As structural analysis using mass
spectrometry requires fairly high concentrations (µM –nM), reducing the volume during the
sample preparation step actually results in higher concentrations which can be delivered directly
to the mass spectrometer.
We will design and construct two micro-machined devices for lumenal protein purification.
The first will be based on standard electrophoretic (i.e., charge) separation. The goal here will
not be to develop a replacement for 2-dimensional polyacrylamide gel electrophoresis (2D
PAGE), but to optimize the interface between the protein isolation, protein purification and mass
spectrometry steps. The second device to be investigated will be a more elaborate micromachined system which will contain several different sample purification modules. The modules
to be investigated will allow separation of proteins and peptides based on their chemical and
physical properties. In addition, modules which contain immobilized proteases (enzymes which
digest proteins), can be constructed which will allow us to generate peptide maps of the proteins
of interest in-line prior to direct mass spectrometric analysis.
Figure 1. The use of soft X-rays (LIGA) and
plastic materials serving as the substrate has
allowed us to micromachine components with
unprecedented aspect ratios for the proposed
applications. Here is a scanning electron
micrograph of high aspect ratio structures injection
molded from poly(methylmethacrylate), PMMA.
Mass spectrometry has long been a preferred method of analysis due to its speed, sensitivity
and ability to physically separate (by mass) complex mixtures. Recently, a number of uses of
mass spectrometry in gene and protein studies have been proposed. Although mass spectrometry
is theoretically well-suited for such applications, a number of technical and scientific concerns
have limited its use in these areas. To overcome many of these concerns, we will focus on the
development of an appropriate interface between micro-machined devices and the mass
spectrometer which will permit the structural characterization of small amounts of biomaterials.
Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is a popular
analytical method for the characterization of peptides and proteins. This technique involves
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combining the analyte of interest with an untraviolet light-absorbing molecule (the matrix), cocrystallizing this mixture on a sample plate, and analysis of via desorption and ionization using an
ultraviolet laser. Electrospray ionization mass spectrometry (ESI-MS) is a complementary
method to MALDI-MS, and involves flowing a solution containing the protein or peptide of
interest through a needle held at a high electric potential. The aerosol produced by this process,
which contains charged droplets of the analyte, is then focussed into the mass spectrometer. Both
MALDI-MS and ESI-MS are proven technologies for peptide and protein analysis; the Limbach
lab has extensive experience with these technologies.
The Limbach lab, in conjunction with the Soper lab, will focus on developing appropriate
interfaces between the micromachined sample preparation devices and the MALDI or ESI mass
spectrometer. We propose to develop a unique sample deposition process that will permit the
sample to be placed on the MALDI sample plate with the appropriate matrix solution directly
from the micromachined device. Our approach is based on a prior macroscale method of sample
deposition.(3) For the ESI-MS studies, we will investigate a variety of different interfaces which
will couple the micro-machined devices to the ESI mass spectrometer.(4) Preliminary work in
this area is already in progress in the Limbach lab and we have identified that the physical
coupling of the ESI transfer device (which is a fused silica capillary) to the micromachined
device is an area in need of more study. Dr. Soper's group generates micromachined devices
from poly(methylmethacrylate), PMMA; hence, unique coupling methods are available that
cannot be had with the traditional glass-based micromachined devices.
The students involved in this research project will be exposed to a variety of interrelated
technologies and disciplines. The students will learn classical protein chemistry techniques, will
be exposed to new instrumental methods of analysis, and will have the ability to design an
experimental protocol that is optimized for the analysis of low amounts of proteins isolated from
our organisms of interest. By combining the protein chemistry with the technology
developments, these students will be able to develop more efficient preparation or analysis
methodologies, and these students will develop an understanding and appreciation of the role of
technology in biochemical advancements.
Number of IGERT apprentices to be recruited and probable home departments: Two--one
from Chemistry & one from Biological Sciences.
Consistency with the Macromolecular Education, Research & Training theme: The project
requires its students to understand proteins, synthetic polymers used for microfabrication, and
advanced methods for characterizing these macromolecules. The MS-I and MS-II courses are
particularly valuable, for one must learn the state of the art in equipment and methods before
advancing it.
How does the project form a vector cross-product of existing research themes by the
participants?
Existing research directions. Bricker's group has been involved in the structural characterization
of membrane protein complexes for fifteen years Limbach's group has focused on developing
new analytical mass spectrometric approaches for the analysis of complex biomolecule systems.
Bricker and Limbach have recently begun a collaborative project aimed at developing new mass
spectrometric approaches for the analysis of hydrophobic peptides. During that collaborative
project, these investigators have found that analysis bottlenecks include the lengthy sample
preparation steps and the poor sensitivity of conventional sample preparation techniques.
Soper's group has focused on fabricating micro-instrumentation using PMMA instead of
traditional glass-based substrates. His group has also used these devices for high-throughput
analysis of DNA for sequencing and diagnostic applications.
New research direction. This project will bring together for the first time this diverse group of
investigators to develop new methodologies and technologies from the "ground up". The
micro-instrumentation will be designed with the specific needs of this research project in mind.
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The mass spectrometric characterization of proteins and peptides delivered from the micromachined devices is a naturally extension of the current collaborative work between Bricker and
Limbach. This research project will also lead to spin-offs in other areas of biomolecule
characterization including new instrumentation for genotyping or other proteomics interests.
How do students benefit from the team-oriented research, beyond what would be available
to them from either advisor separately? Biological mass spectroscopy is a very hot area for
employment. The student from biological sciences will gain a far deeper understanding of the
mass spectroscopy craft from Professor Limbach then would be obtainable simply by pushing
buttons on the instrument. Similarly, students involved in the microfabrication aspects will
have to apply them to a complex biological system that they might never consider without
Professor Bricker's expertise--and reconcile their results with the large knowledge base for that
system.
Briefly describe the support level available to each individual faculty or off-campus
participant (i.e., without IGERT) The LSU faculty involved are all independently supported
for research in related fields. Together, they hold 8 major grants of about $800,000/year. This
ensures a stable environment for the IGERT students, including healthy exchange of skills and
ideas with postdocs and graduate and undergraduate students.
Interdisciplinary strengths of the team project: Although Limbach and Soper were both
trained as analytical chemists, Limbach has focused on biological mass spectrometry and Soper
concentrates on developing new instrument for molecular separations. Bricker's background is
in microbiology; he brings the more classical biochemist approach to this project. Bricker and
Limbach collaborate currently on membrane protein characterization. These investigations led
to this proposed project.
Commitment of faculty & off-campus participants to work side-by-side with apprentices:
Bricker, a full professor within the Department Biological Sciences, also maintains an active
personal research program and interacts daily with his laboratory members. He is excited about
the possibility of personally working side-by-side on a project of 2-6 weeks with students in the
“master-apprentice” model presented in this IGERT proposal. This time commitment would be
fulfilled during winter or spring breaks. Soper is currently an associate professor of Chemistry.
He has been actively and personally involved in the micromachining and the fabrication of
micro-instrument platforms for the past several years as part of NIH-sponsored research. The
students involved in the micromachining aspects of the project will utilize the CAMD
synchrotron storage ring and related facilities; we are well-equipped to carry out all phases of
the micromachining tasks. Soper will also incorporate both students involved in this project
into his group's normal group meeting schedule so that the students can present their results in a
formal setting. Limbach is presently and assistant professor of Chemistry. His involvement
will include training the students on the necessary instrumentation, assisting the students with
the micro-instrumentation–mass spectrometry interface designs, and working with the students
on the protein characterization studies. This can easily lead to a small project and report with a
new apprentice. Outside of the project meetings, Limbach will meet regularly with the students
to assess their progress and to guide these students on appropriate upcoming experiments.
References:
(1) "The RNA world" edited by Raymond F. Gesteland, John F. Atkins. Cold Spring Harbor, NY : Cold
Spring Harbor Laboratory Press, 1993. 630 p.
(2) Prevost, M., Vu, M., Frankel, LK. and Bricker, T.M., in preparation.
(3) a.Hensel, R. R.; King, R. C.; Owens, K. G., Rapid Commun. Mass Spectrom. 1997, 11, 1785-1793. b.
Axelsson, J.; Hoberg, A.-M.; Waterson, C.; Myatt, P.; Shield, G. L.; Varney, J.; Haddleton, D. M.;
Derrick, P. J., Rapid Commun. Mass Spectrom. 1997, 11, 209-213.
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(4) a. Valaskovic, G. A.; McLafferty, F. W., J. Am. Soc. Mass Spectrom. 1996, 7, 1270-1272. b.
Mazereeuw, M.; Hofte, A. J. P.; Tjaden, U. R.; Greef, J. v. d., Rapid Commun. Mass Spectrom. 1997,
11, 981-986. c. Bateman, K. P.; White, R. L.; Thibault, P., Rapid Commun. Mass Spectrom. 1997, 11,
307-315. d. Hannis, J. C.; Muddiman, D. C., Rapid Commun. Mass Spectrom. 1998, 12, 443-448.
D2. Hairy Rod Polymers in Supercritical Fluids
This project will: enable two IGERT students to synthesize, characterize and process materials
based on
rod-shaped polymers using environmentally safe supercritical fluids.
Primary Faculty co-Advisors:
William H. Daly, Chemistry Department (Synthetic Polymer Chemistry)
Maciej Radosz, Chemical Engineering Department (Thermodynamics)
Paul S. Russo, Chemistry Department (Polymer Physical Chemistry)
Off-campus Participant: Gerhard Wegner, Director, Max Planck Institut für Polymerforschung,
Mainz, Germany
Technical Proposal: Nature often selects rodlike macromolecules for structural applications, an
important example being collagen fibers in skin and tendon. Human attempts to design structures
with rodlike polymers are primitive by comparison. One of the main problems is that most
rodlike polymers do not melt; unlike polyethylene and many other common polymers, it is
usually impossible to mold rodlike polymers at high temperature. Rodlike polymers such as
Kevlar are usually processed from solvents, often rather harsh or environmentally damaging
solvents to overcome their poor solubility. The solubility of rodlike polymers improves if they
contain spacer groups or flexible “side chains”. Here is depicted a “hairy” rodlike
macromolecule with flexible side chains.
Side View
End View
Hairy Rodlike Macromolecule
At sufficiently high concentrations, rodlike macromolecules spontaneously form liquid crystals,
which can be processed into very strong fibers or films. Films can be produced by successively
dipping a plate into a liquid on whose surface floats a single layer of carefully spread rods
(Langmuir-Blodgett method).
Production of three-dimensional, solid materials is not so highly developed, but such
materials would offer many desirable features: light weight, directed strength, stability to high
temperature and harsh chemicals and, possibly, directed response to stimulus. The objective of
our IGERT team will be to understand and process hairy rodlike macromolecules into solid
materials using supercritical fluids, including environmentally benign carbon dioxide. A
supercritical fluid is a solvent that is neither gas nor liquid; like a gas, it expands to fill the
available volume, but its density lies closer to that of a liquid. The most common example is
carbon dioxide at pressures exceeding about 60 times atmospheric and temperature exceeding
30oC (Although CO2 is a greenhouse gas, the CO2 is easily recovered and recycled, as will be the
other supercritical fluids used in our work). Supercritical fluids do not have any surface tension,
so evaporating and recovering the solvent does not generate the capillary forces that would
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destroy materials during processing. The research requires synthesis, molecular and
thermodynamic analysis that underlies processing from supercritical fluids, and characterization
of solid materials and precursor solutions.
Synthesis. Some of the most appealing macromolecules for fundamental study are based
share the same general structure, but the
on a synthetic polypeptide structure, shown below. Proteins
substituent groups (indicated by R, R', R"
O
H
O
H
R'
H
etc.) appear in a particular sequence in
C
N
C
N
proteins. Here only a few special R groups
N
C
C
will be used. The resulting polypeptides
H
R"
R H
H
O
O
will generally be soluble in organic solvents
n
such as supercritical
propane; unlike proteins, they are not usually soluble in water. To enhance the solubility of the
peptides in supercritical CO2, some of the substituent groups might be fluorinated alkyl chains. A
typical polypeptide for this study poly(-carbobenzoxy-L-lysine-co-decenyl-glutamate), is shown
below (without fluorination).
The rodlike character of the polypeptides
CBZ HN
derives from their tendency to wind up
O
H
O
H
like helical springs, rendering them
H
C
N
C
N
remarkably stiff in terms of bending
N
C
C
rigidity.
By polymerization of the
H
H
H
O
O
corresponding N-carboxyanhydrides, the
n
polypeptides are available in gram
O
C
CBZ HN
O
quantities with good size uniformity.
6
Thesepro
These properties make polypeptides excellent model systems for studying the fundamental
characteristics of other rodlike polymers that may ultimately prove more practical in some
applications, such as the poly(phenylenes) and functionalized cellulosics being developed at the
Max Planck Institute.
The poly(-carbobenzoxy-L-lysine-co-decenyl-glutamate) shown above would be
expected to form liquid crystals. The backbone amino acid used easily undergoes thermal
transitions from helix to coil, making the polymer and structures made from it sensitive to
temperature change. The decene double bond allows this polymer to be crosslinked efficiently
and rapidly with catalysts developed by Grubbs' group (Schwab, 1995) as shown below (at left).
O
N
H
COO
O
6
N
H
COO
6
isotropic gel
cat
O
N
H
COO
= crosslink
O
6
N
H
COO
6
liquid crystal gel
Covalent networks of rods can be prepared from solutions as shown above (at right) for randomly
oriented (isotropic) and aligned (liquid crystalline) cases. The March ACS National Meeting in
San Francisco will feature an entire symposium devoted to production of aligned networks,
reflecting the excellent control over materials properties such systems provide. After trying
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Pressure
14
T
UCS
slower and harsher methods of crosslinking developed elsewhere,(Kishi,1990) the above
chemistry was developed by former LSU student Drew Poche’ (Daly advisor, with an assist from
Russo). It works quickly and reliably, but needs to be tested further and optimized. Mild
discoloration must be reversed, and we must confirm that alignment is preserved when
crosslinking liquid crystalline solutions. The student performing this research will learn in an
integral fashion organometallic catalysis, spectroscopic techniques, light microscopy and small
angle X-ray scattering for confirmation of alignment. Even with a somewhat smaller skill set, the
student would be highly marketable: Poche' flirted with academia before taking a job in
polyolefin characterization.
Molecular Analysis in Supercritical Fluids. The rodlike helical structure of the
polypeptides is not guaranteed to exist in all solvents; it must be confirmed in each, including the
supercritical fluids. Also, the state of aggregation of the rods needs to be assessed. These
molecular scale analyses will begin in liquid solvents (as a control) and progress with student and
instrumental capabilities towards supercritical fluids. The primary tool will be static and dynamic
light scattering, which can measure the size of molecules as a function of their mass. Such size
vs. mass relationships provide shape information (size increases linearly with mass for rods,
whereas for spherical solids it increases as the cube root of mass). In simple solvents, this
information is rapidly available from gel permeation chromatography/light scattering/viscosity
(triple detector) measurements increasingly used in industrial labs. The student must master
thermodynamics, optics, and data analysis to perform such research, reinforcing and motivating
classroom instruction. We do not contemplate triple detector methods for supercritical fluids.
Instead, dynamic light scattering will be correlated with the chromatographic results in simple
solvents and then developed for supercritical fluids. We are now upgrading our NSF-funded
dynamic light scattering facility; the IGERT student will make further alterations to enable
measurements in supercritical fluids following recent progress here (Chan, in press) and
elsewhere.(Szydlowski, 1998). This student will acquire the different skills of both Radosz and
Russo.
Thermodynamic Analysis of Processing from Supercritical Fluids. The crucial
challenge in processing polymeric materials from solution is how to separate the solvent. Subcritical liquid solvents are separated from polymer by evaporation or by dilution with an
antisolvent. The supercritical solvents, on the other hand, can be separated by rapid expansion,
which is much faster and allows far greater flexibility in controlling the shape and size of
particles and pores. Rapid-expansion separation is applicable to supercritical-fluid and polymer
pairs that mix completely. The pairs that do not exhibit complete miscibility can be processed
using a hybrid approach: we dissolve the polymer in a sub-critical liquid, but instead of
evaporating the solvent, we pressurize the solution with a supercritical antisolvent to form the
solid material.
In all these approaches, the final material morphology sensitively depends on the phasediagram path and the rates of changing the pressure, temperature, and composition from the initial
solution to the solvent-free material. Optimization of this path is the key to making desirable
materials reproducibly. This, in turn, calls for understanding the solution phase behavior at highpressures. While the phase behavior of polypeptide solutions in sub-critical liquid solvents is
relatively well established, the behavior of rigid polypeptide solutions in supercritical and nearcritical solutions is completely unknown.
In separate but related projects, however,
we have studied the phase behavior of flexible
L
polymers and copolymers, such as polyolefins, in
supercritical fluids. We found that supercritical
fluids can significantly shift the crystallizable
U-LCST
solid-liquid and fluid-liquid transitions, relative to
liquid solvents. We learned how to relate these
LL
LCS
T
LL
LLV
VL
LLV
UCEP
LCEP
VL
Temperature
February 2003
shifts to the solvent and polymer structure. Appearing at right is a generic example of the
pressure-temperature phase diagram at constant composition that captures the behavior of many
amorphous-polymer solutions in supercritical and sub-critical solvents: Here, L stands for liquid,
V for vapor, UCST for the upper-critical solution temperature, and LCST for the lower-critical
solution temperature. The different U-LCST pairs of phase boundaries correspond to different
polymer-solvent mixtures. These mixtures become completely miscible at pressures above the
phase boundary. We need such a complete map of phase boundaries as a function of polymer
characteristics (molecular weight, side-chain density and distribution, and rigidity) in order to
design a solvent-separation paths and rates for material processing from supercritical fluids.
An example of relevant research is taken from our phase behavior and particle-formation
study (Yeo, 1995) on rigid-rod aromatic-polyamide materials, similar to Kevlar. The hybrid
approach (supercritical antisolvent precipitation) was used, where the primary solvent was a subcritical liquid (for example dimethylsulfoxide = DMSO) and the antisolvent was a supercritical
fluid (for example CO2). The main result was that aromatic-polyamide type rigid-rods can be
made miscible in DMSO-like aprotic solvents, and processed with supercritical CO2, by attaching
spacer groups along the aromatic-polyamide rods to reduce hydrogen bonding, and hence
improve miscibility. Our proposal to attach the hairy side chains to the polypeptide rods rests on
a slightly different premise–we want to improve their chemical affinity to the solvent and
sidechain mixing entropy– but it should produce a similar result: the rigid polypeptide rods
should become miscible in supercritical fluids.
The second example confirms this hypothesis and proves the feasibility of this project.
The polymeric solute in this preliminary experiment is a poly(stearyl-glutamate) (PSLG) having a
mass of about 9000/mol. The solvent is propane. The pressure-temperature phase diagram for a
5 % solution shown at right suggests that PSLG is immiscible with propane below about 50oC
(the area below the curve is a two-phase region). The good news is that propane seems to be a
good solvent for PSLG at higher temperatures as long as pressure is high enough, at least 300 bar
in this case. Such pressures are easily achieved in Radosz' lab. This phase diagram also suggests
creative processing approaches: for example, dissolve PSLG in sub- or supercritical propane at a
high pressure (say above 50oC and 300 bar) and then precipitate by expansion, cooling or both.
In addition to probing fluid-liquid and solid-liquid transitions, the team will look into isotropicliquid crystal transitions and other solution properties. The chemical-engineering student
working on this project will learn the high-pressure techniques and thermodynamics underlying
high-pressure phase transitions in macromolecular systems, including the elements of modeling
using equations of state. Industrial advisors confirm that these skills are highly marketable.
Characterization of Materials. Isotropic covalently crosslinked gels can be
characterized by a correlation length, which may be thought of as the average distance between
crosslinks. This will be determined from small angle X-ray scattering (the variation of intensity
with scattering provides the necessary information). By the same method, liquid crystalline gels
can be assessed for the degree of order that remains after crosslinking. To assess the damage that
may occur during supercritical fluid extraction, the same parameters must be measured in the
precursor solutions.
Number of IGERT apprentices to be recruited and probable
home departments: Three--one from Chemistry & two from
Engineering (theory & experiment).
Consistency with the Macromolecular Education, Research
& Training theme: Macromolecules will be made from small
monomers distantly related to biopolymers, characterized
under challenging conditions no biopolymer ever saw, and
processed into novel materials.
How does the project form a vector cross-product of existing
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February 2003
research themes by the participants?
Existing research directions. Russo’s research group has a longstanding program concerned with
the dynamics of rodlike polymers in normal liquid solvents, such pyridine or 98% sulfuric acid.
Daly’s research group has a well-established program in the synthesis and modification of
biosimilar macromolecules, including semi-rigid polymers. Radosz’ group has expertise in
behavior of macromolecules in supercritical fluids. The Wegner group at the Max-Planck
Institute in Germany is world-renowned for the development of hairy rod polymers as novel
materials.
New research direction. This is a first attempt to characterize and process hairy-rod polymers in
supercritical fluids. The probability of success, however, is high because this team has a unique
set of resources and expertise to synthesize, characterize and process hairy rod polymers in
supercritical fluids.
How do students benefit from the team-oriented research, beyond what would be available
to them from either advisor separately? The immediate benefit from the student’s
perspective is a larger pool of multidisciplinary expertise on which to draw. The long-term
benefit is that such research, which cannot now be performed efficiently in any single group,
places the student at the technical forefront. Our goal is to train a student who can “do it all” in
this new field. Such a student would occupy a unique niche in the academic world, should he or
she choose that path. With a broad background in synthesis, scattering, microscopy and highpressure methods, the student would do equally well in an industrial environment.
Briefly describe the support level available to each individual faculty or off-campus
participant (i.e., without IGERT) The LSU faculty involved are all independently supported
for research in related fields, with a "net worth" of 8 grants totaling $700,000/year. Truly
excellent resources await the student(s) who visit the MPIP in Mainz. This ensures a stable
environment for the students, including healthy exchange of skills and ideas with postdocs,
graduate students and undergrads.
Interdisciplinary strengths of the team project: Daly and Russo are at opposite ends of the
wide spectrum of scientists that populate modern chemistry departments, while Radosz is an
engineer. Daly is trained as an organic polymer chemist; typical activities in his group include
synthesis of monomers, polymerization into macromolecules, purification, and various spectral
identification techniques and simple analytical methods such as viscosity. Russo is trained in
physical chemistry; typical activities in his group include preparation of solutions, light
scattering and fluorescence photobleaching recovery measurements for physical properties like
mass or diffusion, optical microscopy, optical design, and writing custom programs for
instrument interface or data analysis. Radosz is an engineer. Typical activities in his group
include high-pressure fluidics, polymer fractionation, developing thermodynamic theory and
computer simulations of phase separation.
Commitment of faculty & off-campus participants to work side-by-side with apprentices:
Russo enthusiastically commits one month full time to the side-by-side participation during
winter holiday, between fall and spring semesters. We will dive right in to light scattering of
polymers, but under conditions of high temperature not high pressure. The purpose will be an
instructive miniproject involving molecular weight and diffusion measurements of one or two
carefully fractionated polyethylene supplied by Dow Chemical. The polymers are not those
ultimately to be studied, but this project provides a good opportunity to learn some basic
experimental light scattering, face some difficult challenges (high temperature requires
resourcefulness), strengthen industrial contacts in the key polyolefin area, and write a report—
all in a reasonable time frame. Daly enjoys working with undergraduate students during the
summer. This summer four undergraduate students worked with him on various aspects of
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February 2003
polymer grafting and had the opportunity to learn organic synthesis and polymer modification
techniques along with the typical approaches to structure identification. One of the polymers
studied is used in a commercial pull-trusion process; future work will involve interaction with
industrial scientists to ascertain if a concurrent pull-trusion/grafting process would yield
superior composites. The project will give the students an opportunity to interact with an
industrial processing laboratory and introduce them to some of the applied aspects of polymer
science. Radosz also enthusiastically commits a total of at least one full month to the side-byside lab and computational work with the student. He will start from the well-established highpressure experimental approaches on polypeptide solutions in propane. Next, he will extend
these experiments to other solvents, such as CO2, and attempt to model the effect of variable
rigidity on the phase behavior with equations of state. These faculty have worked together for
several years on other projects [Chan, Poche' 1]. They often serve on student and university
policy committees together, helped design the new curriculum, and have an excellent working
relationship. If one must be gone for a short while, another could easily “look after” the
student, offering general advice and clarifying the project vision but with less specific technical
expertise.
References:
Chan, A. K. C.; Russo, P. S.; Radosz, M. “Fluid-Liquid Equilibria in Poly(ethylene-co-hexene-1) +
Propane: Light Scattering Probe of Cloud-Point and Spinodal Pressure and Critical Polymer
Concentration” Fluid Phase Equilibria 1999, submitted.
Kishi, R., Masahiko, S., and Tazuke, S. "Liquid Crystalline Polymer Gels. 1. Cross-linking of Poly(
benzyl-L-glutamate) in the Cholesteric Liquid Crystaline State" Macromolecules 23:3779-3784, 1990.
Poche', D. S., Daly, W.H., Russo, P. S., “Synthesis and Some Solution Properties of Poly(-stearyl,L-glutamate)” Macromolecules, 1995, 28, 6745-6753.
Poché, Drew S., Thibodeaux, Stefan J., Rucker, Victor C., Warner, Isiah M., Daly, William H.
"Synthesis of Novel -Alkenyl-L-glutamate Derivatives Containing a Terminal C-C Double Bond to
Produce Polypeptides with Pendent Unsaturation", Macromolecules, 1997, 30, 8081-8084.
Schwab, P., France, M.B., Ziller, J.W.; Grubbs, R.H "A series of Well-defined Metathesis Catalysts-Synthesis of [RuCl2(=CHR')(PR3)2] and its Reactions" Angew.Chem.Int.Ed.Engl. 34(18), 20392041, 1995.
Szydlowski, J.; Rebelo, L.P.; Wilczura, H.; Dadmun, M.; Melnichenko, Y.; Wignall, G.D.; Van Hook,
W.A.: "Comparison of SANS and DLS hydrodynamic correlation lengths for a
polystyrene/methylcyclohexane solution in the vicinity of temperature or pressure induced critical
demixing," Physica B. Condensed Matter, 1998, 241/243, 1035-1037
Yeo, S. D.; Debenedetti, P. G.; Radosz, M.; Schmidt, H-W. "Supercritical Anti-Solvent (SAS) Process for
Substituted Para-Linked Aromatic Polyamides: Phase Equilibrium and Morphology Study"
Macromolecules 1993, 26, 6207 and 1995, 28, 1316.
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