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Radiospectroscophy of Bio-Products

#0412


Development of Noninvasive Contact-free Approaches for Monitoring the Production Stages and Controlling the Product Quality by Using Radiospecrtoscopy Methods (NMR, EPR).

Tech Area / Field

  • BIO-CHM/Biochemistry/Biotechnology
  • BIO-MIB/Microbiology/Biotechnology
  • BIO-IND/Industrial Biotechnology/Biotechnology
  • BIO-RAD/Radiobiology/Biotechnology
  • MED-DRG/Drug Discovery/Medicine

Status
3 Approved without Funding

Registration date
13.06.1995

Leading Institute
State Research Center for Applied Microbiology, Russia, Moscow reg., Obolensk

Collaborators

  • University of Wyoming / Department of Animal Science, USA, WY, Laramie

Project summary

The problem on the improvement of the techniques for measuring the major biotechnological parameters, influencing the quality of final product, is one of the most important in the study and optimisation of the processes of biotechnological manufacturing NMR and ESR magnetic radio spectroscopy methods (MR) are widely applied in the studies of various biological objects, but up to now have not been used for the solution of biotechnological problems, though, as it was demonstrated by our preliminary results, they have many advantages for the development of efficient systems of control of industrial processes and product quality.

The major goal of the project proposed is the development of noninvasive contact-free approaches for monitoring the production stages and controlling the product quality by using radio spectroscopy methods (NMR, EPR).

The execution of the following steps seems to be reasonable for the achievement of this goal:

- to perform NMR relaxation analysis of live lyophilized vaccinees and to reveal amplitude - relaxation characteristics of the samples, that will allow one to determine rapidly the residual moisture of the preparations;

- to develop methods for determination of non-freezing water quantity, crystallization, vitrification and melting of the solutions of potential stabilizaters by using temperature dependencies of NMR amplitude - relaxation parameters of frozen solutions and to develop methods for determination of non-freezing water quantity, crystallization, vitrification and melting of the solutions of potential stabilizaters by using temperature dependencies of NMR amplitude - relaxation parameters of frozen solutions;

- to study free-radical transformation regularities and their correlation with the degradation and inactivation of biological material as compared to NMR-relaxation and electronic microscopy data and to develop a many - factor model of dry biopreparation inactivation and to use it for studying comprehensively the mechanism of lyophilized preparation destruction as well as for the prediction of product preservation;

- to investigate cell physiological state for cultivation by using NMR methods (water exchange rate across membranes, intercellular pH, osmotic pressure), to bring to light culture "stationary" NMR - criteria and to perform the analysis of correlation between bacterial cell physiological state of the bio-mass and technical liquid and their tolerance to freezing and drying;

- to carry out the investigation of solid nutrient medium gel bases for the determination of NMR parameters, which are the most sensitive to natural and synthetic gel structure and growth properties.

It is suggested to use different ESR and NMR methods (1-H, 31-P NMR spectroscopy, NMR-relaxation, ESR spin-label ) in accompany with other biophysical methods (electron spectroscopy) and traditional microbiological and biochemical methods too. For determination and quantity evaluation of correlation between RS parameters of dry samples and biological activity of the preparation mathematical statistics and many-factory methods will be used.

The following results are expected to be received:

- test-system for the evaluation of bacterial cell concentration, physiological state of cells and cell damage during cultivation will be designed on the base of the parameters determined by radio spectroscopy methods. The test-system will be used for the analysis of cell response to various damaging factors during biotechnological procedures.

- a computerized method for determination and control of moisture content in lyophilized biopreparations and dry nutrient media based on NMR-relaxation will be developed. Water content limit in dry protein hydrolyzates, upper which water operates as a solvent, will be determined. The contact-free method will provide rapid determination of moisture content with high accuracy;

- NMR relaxation method for quantity determination of unfreezable water, definition of temperature crystallization and phase transition in water solutions potential stabilizers will be developed;

- mathematical model of FR formation and death during storage of dry preparations will be constructed and method for evaluation of the efficiency and stability prediction {activity preservation) of lyophilized biopreparations during storage based on EPR kinetics of free-radical transformation will be developed;

- investigation of the polymers entering into the composition of solid nutrient media as gel basis will be performed by NMR methods, NMR parameters, sensitive to the structure and gel-formation properties of natural and synthetic polymers will be determined;

- investigation of the role of water and bivalent cations in the formation and stability of triple complexes (polynucleotide - metal ion - phospholipid vesicles and micelles from natural and synthetic phospholipid) will be carried out, the hypothesis of the existence of nucleotide - phospholipid - recognition will be verified.

The project participants are interested in cooperation with foreign partners for the exchange of experience and information as well as for practical study of specialists.


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