Two Dimensional Protein Arrays
Two Dimensional Protein Arrays with Regular Structure for Analytical Biotechnology
Tech Area / Field
- CHE-POL/Polymer Chemistry/Chemistry
3 Approved without Funding
ITEF (ITEP), Russia, Moscow
- Research Center of Molecular Diagnostics and Theraphy, Russia, Moscow
- BioGuard Technologies, Inc., USA, WA, Richland\nUniversity of Illinois At Urbana-Champaign / Department of Physics, USA, IL, Urbana\nNew Horizons Diagnostics Corporation, USA, MD, Columbia\nLouisiana Tech University / Institute for Micromanufacturing, USA, LA, Ruston\nEmory University / Department of Chemistry, USA, GA, Atlanta
Project summaryProject Title: Two Dimensional Protein Arrays with Regular Structure for Analytical Biotechnology
Project Manager: Ignatuk Tatyana Evgenievna
Participating Institutions: State Science Center of the Russian Federation-Institute for Theoretical and Experimental Physics; Russian Science Center of Molecular Diagnostics and Treatment
Total Project effort (person*days): 15928
Total Project effort of weapon scientists and engineers (person*days): 8480
Estimated total cost of the project (US$): 620000
1. Description of the problem:
Progress made in the area of enzyme engineering, immunology, electrochemistry, microelectronics, and nanotechnology has promoted the onrush of biosensor technology, giving ability to develop sensor-based system, with potentials of multilevel selection of biological objects. One of the main tasks when carrying out the assay is to perform specific localization of the investigated object on the sensor surface. Protein arrays exhibit the needed functionality required for such sensors, since they can provide specific sorption of the analyzed objects and hence perform the first level of selection. The minimal requirement for such arrays is dense arrangement of binding sites, while the optimum performance can be achieved with two dimensional protein arrays where the molecules, providing specific binding, have regular arrangement and are identically oriented. Regular structure of the affined substrate suggests the following advantages: reliable detection of the specific primary signal, substantial automation of the object identification procedure and effective registration of inpidual biological interactions. Moreover, regular protein structure of the affined substrate can be used as a peculiar molecular transducer and amplifier of the primary signal, due to the initiation of cooperative protein effects.
.Actual methodology allows to fabricate functional protein layers and to perform selective immobilization and patterning with micrometer and sub micrometer accuracy, applying such techniques as photolithography [54, 55, 56, 62], micro contact printing , micro fluid- networks , atomic force lithography [59, 60, 63] and dip pen nanolithography , ], but does not solve 2D regular protein arrays fabrication problem.
.Current project is focused on fundamental research activities in 2D highly oriented protein arrays (antibodies matrixes) fabrication and evaluation of acceptability of such arrays in the area of analytical biotechnology. It is implied, that such techniques as Langmuir method of monolayer films formation, self assembling of biologically active functional layers and scanning probe microscopy (SPM) will be used as basic nanotechnological approaches to the problem. Practically, the goal of this project is to develop new scientific base for bio-analytical purposes to be used in fabrication of biosensor devices, where antibodies, enzymes and nucleic acids play the role of the “recognizing” elements. The objective of this project is mainly related to the necessity of currently existent protein arrays improvement along with the development of new effective protein arrays and protein arrays based analytical devices. Adaptation of regular protein arrays will allow to essentially raise sensitivity to the extent of single molecules and cells, decrease analysis time and to increase reliability. Obvious applications of such 2D regular protein arrays based devices are the assessment of epidemiologic situation in built-up areas and agricultural holdings, food substance inspection, and the development of effective and reliable methods of express microbiological analysis, which will allow to avoid consequences of bio-chemical terrorism.
2. Scientific and technical standards
The executing agency has high potentials for carrying out scientific research and applied investigations in this area of science – the following investigations have been performed since recently:
- SR “Reflection–K” and “Reflection–K2” completed according to the State contract KB–92-23 (23.11.93) and KB-53-96 (31.01.96), negotiated with Chemical and Biological Weapon Conventional Problems Committee under the President of the Russian Federation. The objective of these research activities was investigation of SPM capabilities for biological agents imaging, detection and identification.
- ISTC projects 1055 and 1548, related to the formation of new principles for biosensor testing of biologically active compounds.
- ISTC project 1734, related to the development of SPM based methods of microbiological control of water environments.
The execution phase is represented by complex scientific, technical and technological approaches and developments, including the following keypoints:
- actual approaches to protein immobilization on solid surfaces;
- biotechnology of fabrication, purification and certification of the chosen pathogenic agents and correspondent mono and polyclonal antibodies;
- technology for sensors (affined substrates) fabrication on the basic of atomic flat surfaces;
- structural and physical chemistry of polymer films, used for affined substrates fabrication;
- scanning probe microscopy of biological objects, including force mode microscopy and fluid cell microscopy;
- SPM image processing, related to pure surfaces along with surfaces, containing amphiphilic mono and polymer compounds and their complexes with biologically active substances and microbiological objects;
- development of an automated image recognition procedure and SPM database information input;
- development of customized software environments for SPM data processing.
Each of the decribed points includes scientific and technical base, which reflects potentials of the executing agency.
3. Project practicability
Progress made in the area of enzyme engineering, immunology, electrochemistry, microelectronics, and nanotechnology has promoted the onrush of biosensor technology, giving ability to develop sensor-based system, with potentials of multilevel selection of biological objects. Equipment, possessed by the executing agency allows implementing in a full measure the mentioned results in order to investigate possible approaches to two-dimensional well-order, high-oriented protein arrays fabrication.
4. Originality of the proposed scientific and technical approach to the problem
The proposed scientific and technical solution is unique.
5. Interim results
Investigation of new methods for two-dimensional high-oriented (including well-ordered) protein films formation, quality control of these films, and selection of the optimum SPM based analysis schemas will be carried out in the course of the project.
6. Final results
Final result – conclusion about the possibility to fabricate two-dimensional well-ordered high-oriented protein films for analytical biotechnology.
7. Possible applications of the project results
Two-dimensional well-ordered high-oriented protein films can be used for improvement and development of bioanalytical methods and devices.
8. Potentials for further cooperation after the project completion
Potentials for further cooperation after the project completion lie in further development of methods for two-dimensional well-ordered high-oriented protein films fabrication and application of these films in a new generation of biosensor devices which will use regular protein structure of the affined substrate as a peculiar molecular transducer and amplifier of the primary signal, due to the initiation of cooperative protein effects.
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