**5. References**

14 Will-be-set-by-IN-TECH

sclerosis (ALS). Biodistribution kinetic intervals were3h(125I-scFv) and 6 h (125I-TU-20).

TU-20 and its scFv were labeled with 125I and 123I by chloramine-T (with average yield 0,72 and 0,50, resp.). Radiochemical purity and stability was revealed by gel filtration (decrease to 80 % and 50 % in two months, resp.) Fragmentation of the labeled antibody and its fragment was estimated by bis-tris gel electrophoresis followed by silver staining and autoradiography

Affinity coupling and RIA adaptation for the specific conditions showed 10-30 % preserved immunoreactivity of the labeled compounds. Otherwise, these methods carry out quite high

In vitro studies performed on mice brain slices confirmed several important assumptions. The antibody is preferentially bound in the layer of Purkinje cells in the cerebellum. SPECT camera in vivo experiment deals with these results: activity bound in scFv is primarily distributed to

Distribution images of the labeled TU-20 provides ambigous because the substance is accumulated in the chest and ventral part and image resolution do not afford more detailed biodistribution identification. However, it is known from previous biodistribution preparative

In vivo experiments were focused on investigation of the blood clearance and organ distribution of the radiolabeled TU-20 and scFv fragment in mice. Let´s show especially the results from scFv biodistribution study in preference. It was verified that the major part of activity, according to the amount of the labeled scFv fragment, was eliminated from blood during 2-3 hours. Minor part of activity, according to the amount of the labeled scFv fragment (0,5 - 1,0 %), was kept in the blood for some days. The value t1/2*<sup>α</sup>* for 125I-labeled scFv fragment was calculated as 2,3 h and the t1/2*<sup>β</sup>* was estimated as 62,4 h. The half-life for overall

In comparison, we found that the 125I-labeled scFv fragment uptake in thyroid gland appeared much lower than for Na125I, as expected. The t1/2*<sup>α</sup>* value for 123I-labeled scFv fragment was calculated as 1,4 h, but the long phase elimination half-life t1/2*<sup>β</sup>* was not estimated due to short half-life of the isotope 123I. The radiolabeled scFv fragment passed in general through the digestive tract (stomach and intestine) and finally was eliminated through kidneys in

TU-20 and ScFv TU-20 showed suitable properties for further investigation in animals which are genetically modified mutants with the ALS (Amyotrophic Lateral Sclerosis). Comparing biodistribution experiments in modified organism confirmed expected behavior. The most significant biodistribution differences occured in the area of the limbs and caudal part of spinal

Finally, as I can summarize, TU-20 and its scFv fragment were successfully labeled with radioiodine 123I and 125I, and, subsequently, the biochemical and analytical characteristics were investigated. Biological properties of the radiolabeled TU-20 and its scFv were evaluated

discrepancy and it will be neccessary to provide further optimising search.

the thyroid gland and digestive tract, then passes quickly through kidneys.

study that activity is distributed in lung, heart, liver, stomach and colon in first 6 h.

(Heiman-Patterson T.D., 2005) (Naini A., 2007)

(over 95 % of radioactivity bound in the substances).

elimination of Na125I from blood was 4,5 h.

preference.

cord and spine.

in vivo by biodistribution studies.

**3. Conclusion**


**23** 

*Russia* 

**Gel Electrophoresis as a Tool to Study** 

Lev Elkonin, Julia Italianskaya and Irina Fadeeva *Agricultural Research Institute for South-East Region* 

**Polymorphism and Nutritive Value of the** 

**Seed Storage Proteins in the Grain Sorghum** 

Seed storage proteins of cereals constitute the basis of mankind nutrition. However, climate changes, especially, increased droughts that are distinctly observed in many regions all over the globe, hamper sustainable production of traditional cereals, such as wheat, maize and barley, and dictates necessity to cultivate drought resistant and heat tolerant crops. Among these crops, the grain sorghum, owing to its ability for sustainable grain production in conditions of minimal level of precipitation, takes one of the leading places. However, application of sorghum grain for food and feed purposes is limited by its relatively low

One of the reasons of poor nutritive value of sorghum grain is the resistance of its seed storage proteins (kafirins) to protease digestion. Kafirins are alcohol-soluble prolamin proteins making up to 80% of endosperm sorghum proteins (Hamaker et al., 1995). As well as other prolamins, sorghum kafirins contain high levels of proline and glutamine and are deposited in protein bodies of endosperm cells during kernel development. According to differences in solubility in aqueous *tert*-butanol solutions, molecular weight, structure and immunochemical similarity to zeins (maize prolamins) the kafirins were classified into α-, βand γ-kafirins (Shull et al., 1991; for review, see: Belton et al., 2006). The α-kafirins are highly hydrophobic prolamin proteins (soluble in 40-90% aqueous *tert*-butanol solutions), they comprise 66-84% of total kafirins, depending on the endosperm type (vitreous or opaque). By SDS-PAGE the α-kafirins usually are resolved into two proteins, 25 kDa and 23 kDa. The γ-kafirin accounts for 9-21% of total kafirins depending on the endosperm type (Waterson et al., 1993). According to immunochemical data, the γ-kafirin is a protein with molecular mass of 28 kDa (Shull et al., 1991) although the sequence of the γ-kafirin gene corresponds to the protein with molecular mass of about 20 kDa (De Barros et al., 1991). The β-kafirin, in different endosperm types, accounts for about 7-13% of the total kafirins, and is resolved by the SDS-PAGE into three bands of 20, 18 and 16 kDa (Shull et al., 1991; 1992) or produced one band of 20kDa (El Nour et al., 1998); such variability, perhaps, is due to genotype

One of the main characteristic features of kafirin proteins is their ability to form olygo- or polymers of high molecular weight. These oligomers comprise α- and γ-kafirins linked

**1. Introduction** 

differences.

nutritive value in comparison with other cereals.


URL: *http://www.sciencedirect.com/science/article/pii/S0969804307002813*

Vijayalakshmi, M. A. (1992). Histidine ligand affinity chromatography, *in* A. Kenney, S. Fowell & J. M. Walker (eds), *Practical Protein Chromatography*, Vol. 11 of *Methods in Molecular Biology*, Humana Press, pp. 33–44. 10.1385/0-89603-213-2:33. URL: *http://dx.doi.org/10.1385/0-89603-213-2:33*
