1985 — 1986 |
Smith, George Pearson [⬀] |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Filamentous Phage Physiology @ University of Missouri-Columbia
I plan to investigate the membrane-associated assembly of the filamentous phages, a group of viruses that infect cells carrying the F fertility factor. My initial approach has two parts: development of simple assays for the stages of virus assembly, and construction and isolation of viral and host-cell mutants that affect the assembly process, so that the specific step affected can be identified with the stage-specific assays. This work should result in a much clearer picture of the components involved in assembly, and set the stage for a deeper study of the process in vitro. If it turns out to be feasible to assemble infectious viruses from naked DNA in vitro, this might be the basis of a system for cloning very large pieces of foreign DNA. I also plan to study other ways in which knowledge of filamentous phage physiology might be expoited to devise new ways of manipulating and analyzing cloned DNA.
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0.915 |
1989 — 1993 |
Smith, George Pearson [⬀] |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Filamentous Fusion Phage @ University of Missouri-Columbia
Filamentous fusion phage display the amino acids coded by a cloned DNA insertion the surface of an infectious virus particle. Phage bearing a particular foreign determinant can be affinity-purified with antibody, making it easy to purify clones that are as rare as 1 in 100 million in the original mixture. Methods will be devised for constructing fusion-phage "libraries" containing 100 million different foreign DNa inserts, so that the full potential of the technology can be exploited. The concept of an epitope library exemplifies the uses to which fusion phage might be put. The foreign DNA inserts in this library would be synthetic DNa with random sequence. Billions of short amino acid sequences would be represented in a 100-million-clone library; it is likely, therefore, that it will contain short determinants ("epitopes") recognized by any anti-protein antibody. This idea will be tested with antibodies directed against myohemerythrin, a small protein whose antigenic structure has been intensively studied with synthetic peptides. The results should yield a wealth of new information about the specificity of anti-protein antibodies-- information that touches on the fundamental issue of how the immune system manages to specifically recognize a seemingly unlimited repertoire of different protein antigens. They should also indicate the feasibility of future applications of the epitope library. One possibility is that it could be used to determine the epitope recognized by an interesting antibody--one, say, that confers protective immunity to some disease. This information could then be used to design a synthetic vaccine or immunogen capable of eliciting similar antibodies in other individuals--all without having to clone and analyze the gene encoding the natural antigen.
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0.915 |
1997 — 1999 |
Smith, George Pearson [⬀] |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Epitope Discovery--a New Route to Vaccines @ University of Missouri-Columbia
Bacteriophage T4 serves as a model ~pathogen~ for evaluation a new vaccine development strategy called epitope discovery. The goal is to find artificial mimics of authentic pathogen B-epitopes that can serve as components of an effective vaccine. The source of mimics are large libraries of random peptides genetically fused to the surface of filamentous phage-display vectors. Mouse anti-pathogen antibodies--we call them direct antibodies because they are elicted directly by the pathogen--have been used to affinity select clones out of the phage libraries whose displayed peptides bind them strongly. These peptides are antigenic mimics of the corresponding authentic pathogen epitopes. Are they also immunogenic mimics, in the sense that they elicit an antibody response that cross-reacts with the pathogen itself? That is a vital question in evaluating the promise of epitope discovery for vaccine development, since only immunogenic mimics can have disease-protective value. Accordingly, a panel of antigenic mimics will be assessed for two key components of immunogenic mimicry. First, mice will be hypperimmunized with each of the antigenic mimics and the resulting indirect antibodies titered against both the antigenic mimic and the authentic pathogen epitope. Comparable titers would indicate that the former is a good immunogenic mimic of the latter. Second, mice will be primed with antigenic mimics (long with appropriate T epitopes) to determine if the resulting memory B cells can be mobilized by a challenge with the pathogen itself.
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0.915 |
2000 — 2002 |
Smith, George Pearson [⬀] |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Epitope Discovery--New Route to Vaccines @ University of Missouri-Columbia
"Epitope discovery" is a new way to identify antigenic fragments of pathogen proteins. Starting with a library of tens or hundreds of millions of random fragments of pathogen proteins, anti-pathogen antibodies are used to select fragments that bind particularly tightly to subspecificities within the antibody population. Selection is accomplished with simple microbiological methods by means of phage display technology. The selected peptides, having won a rigorous competition among all the structures in the initial library, have strong credentials as candidate components of synthetic or recombinant peptide vaccines. But are these peptides good "immunogenic mimics"? That is, are they able on their own to induce antibodies that cross-react with the pathogen itself--as they must if they are to protect against disease? This key question will be addressed using bacteriophage T4 as a model "pathogen" (T4 is readily and safely produced in large amounts, allowing cross-reactions to be quantified directly by simple immunochemical techniques). From the results we will learn which intrinsic properties of peptides can be used to predict outstanding immunogenic mimicry a priori, without having to assess it directly in living subjects--a difficult task in the context of actual diseases. Immunogenic mimicry is necessary but not sufficient for vaccine performance, however, since not all pathogen-reactive antibodies actually protect against disease. Using a second model system--the cattle disease caused by the tick-borne, malaria-like parasite Babesia bovis--we will show how antigenic peptides can be screened in vitro for the likelihood of protective value. Taken together, the constellation of innovations to be explored--isolating particularly promising peptides via epitope discovery, assessing them for properties that correlate with superior immunogenic mimicry, and screening them in vitro for the capacity to engender protective effects--promise to deliver vaccine candidates with excellent prospects for efficacy, before a single trial in a living subject must be undertaken.
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0.915 |
2008 — 2009 |
Smith, George Pearson [⬀] |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) |
Small Pretargeting Constructs With Infinite Affinity For Radiochelates @ University of Missouri-Columbia
[unreadable] DESCRIPTION (provided by applicant): Pretargeting is a notable advance in radioimaging and radiotherapy. It allows tumor-avid antibodies and other targeting molecules that have excellent specificity and affinity but poor pharmacokinetic properties to be used in conjunction with small radioactive effectors (e.g., radiochelates) that have superior pharmacokinetic behavior. The key is a bifunctional pretargeting probe, in which the targeting module (e.g., tumor-avid antibody) is linked to another module that specifically captures the radioactive effector. A pretargeting regimen then plays out in two stages. In the first, the bifunctional probe is administered and allowed to home in vivo to the tumor or other target cells or tissue by virtue of its targeting module. Although it may take a day or more for non-targeted probe to clear the body because of its poor pharmacokinetics, it is not radioactive, so the subject experiences no radiation burden and there is no loss of short-lived isotope. Once the probe has cleared, the second stage is implemented: administration of the radioactive effector. It clears from the body rapidly, but during its brief residence some of it is captured by target-bound bifunctional probe molecules via their effector-capture modules. Thus can an effective radioimaging or radiotherapy payload be delivered to the target while imposing a very low background or non- Couples covalently in vivo therapeutic radiation burden on the subject. hel To date, effector-capture modules have been macromolecular. We hope to demonstrate that small Targeting peptides can serve just as well. Using novel phage display module Peptide covalent technology, we will select peptides that couple rapidly, effector-capture selectively, and covalently to small radiochelate effectors with module excellent pharmacokinetic characteristics. Such peptides l retargeti would have important advantages over macromolecular effector-capture modules: (1) They are easy to fuse genetically or couple chemically (in multiple copies if appropriate) to any targeting module; a single generic radiochelate could be used with an unlimited repertoire of stable, non-radioactive bifunctional probes. (2) They can be synthesized chemically, making fully synthetic pretargeting probes possible; mirror-image peptides would capture the opposite radiochelate isomer and be resistant to proteolytic degradation. (3) They will not be immunogenic, and therefore can be used more than once in the same subject. [unreadable] [unreadable] PUBLIC HEALTH RELEVANCE Cancer doctors increasingly rely on radioactive probes that home specifically to a patient's cancer cells. The radioactive probes can be used both to image the cancers for effective diagnosis and to deliver lethal radiation specifically to the cells to help cure the disease, both without invasive surgery. The purpose of this research is to improve and simplify "pretargeting," a new procedure for using probes that can dramatically sharpen diagnostic images and reduce the harmful radiation side-effects patients suffer. [unreadable] [unreadable] [unreadable]
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0.915 |
2009 — 2015 |
Pettey, Dix Smith, George (co-PI) [⬀] Smith, George (co-PI) [⬀] Glaser, Rainer (co-PI) [⬀] Schmidt, Francis (co-PI) [⬀] Hart, Jennifer (co-PI) [⬀] |
N/AActivity Code Description: No activity code was retrieved: click on the grant title for more information |
Prism: Mathematics in Life Sciences @ University of Missouri-Columbia
Intellectual merit of the proposed activity: The overall goal of this PRISM-funded project is two-fold: to recruit mathematically talented undergraduates into science, technology, engineering and mathematics (STEM) disciplines, and at the same time to integrate mathematics more thoroughly into the introductory STEM curriculum, especially in the life sciences. Our program takes advantage of two key undergraduate programs that are already well-established at MU: the Freshman Interest Groups (FIGs) that are centered in the Freshman residence halls, and the Life Science Undergraduate Research Opportunity Program (LS-UROP). A new Mathematics in Life Sciences (MLS) FIG with 20 entering freshman (the MLS scholars) per year will be established in the Discovery residence hall; the students will live together during their freshman year and take an integrated curriculum of three courses developed by the MLS faculty in their first semester. That curriculum, in conjunction with a weekly Proseminar and other FIG activities and services, will create a learning community encompassing both faculty and scholars, with salutary effects on student retention and level of academic engagement. Building on that learning community, students will pair with faculty (both MLS faculty and other professors) for mentored independent undergraduate research projects that will start in the summer under the LS-UROP (with grantfunded internship stipends) and continue through both semesters of the Sophomore year. Admission to the MLS program will be in the form of a scholarship, which will include automatic enrollment in the MLS FIG, the summer stipend, and in some cases academic year funding for all four college years. Although the MLS academic activities proper will end at the end of the scholars' sophomore year, their subsequent progress will be followed closely in order to assess critically the degree to which the program is successful in recruiting students who go on to get a degree in a STEM discipline. The program includes a vigorous effort to recruit students from underrepresented groups, exploiting for this purpose collaborative relationships MU in general and the MLS faculty in particular have establish with high-needs high schools over many years.
Broader impacts resulting from the proposed activity: Mathematics is becoming increasingly integral to all STEM disciplines, especially life sciences. If successful, the MLS program would demonstrate how learning communities like MU's FIGs can be used to integrate mathematics more thoroughly into the introductory STEM curriculum, and enable beginning students to engage in a scientific enterprise in which mathematical facility is a core skill. We are betting that programs like this will be outstandingly successful not only in attracting able students who might otherwise choose non-STEM majors, but also in encouraging those students to complete STEM degrees. The assessment component of the MLS program will provide rigorous measures of the degree to which this aspiration is realized in fact.
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0.915 |