The immune response of chickens immunized with β-N-acetylglucosaminidase was monitored in the egg yolks of the birds using an enzyme-linked immunosorbent assay. Significantly higher levels of specific antibodies were detected in the yolks of the birds immunized with the enzyme when compared with the yolks of a control bird collected over the same period significant differences also were found in the response within the immunized group of birds, indicating individual variability to the injections. Immunoglobulins were isolated from egg yolk after a preliminary purification using alginate to precipitate lipoproteins. A ten millilitre DEAE-Sephacel ion exchange chromatography column resulted in a final product containing 16 mg of IgG with a purity of 60% when 50 mL of an egg yolk supernatant was applied. Specific antibody activity toward the antigens β-lactoglobulin and E. coli lipopolysaccharide was higher in both cases in the isolated immunoglobulin fractions which contained lower purity (40%). Lower antibody activity was observed in the 60% purified fractions. Separation of specific antibodies from non-specific antibodies appeared to occur, possibly due to the given characteristics of the specific antibodies, or due to the differences exhibited by chicken IgG subpopulations. Using metal chelate interaction chromatography, a 10 mL copper-loaded column was able to yield 104 mg of IgG with a purity of 75% when 200 mL egg yolk supernatant was applied. Again, the heterogeneous nature of chicken IgG was illustrated. A comparison of the two chromatographic techniques indicated the advantages of metal chelate interaction chromatography over ion exchange chromatography under the conditions examined. Applications of the chicken IgG isolated by metal chelate interaction chromatography to an enzyme-linked immunosorbent assay for the detection of β-N-acetylglucosaminidase was attempted. Linear relationships were obtained when standard solutions of the enzyme were used in the assay. The results indicate that MCIC-isolated chicken immunoglobulins have excellent potential for use in analytical tests. / Land and Food Systems, Faculty of / Graduate

Diss. (sammanfattning) Stockholm : Karol. inst., 2002. / Härtill 6 uppsatser.

19 Dec 2004. / Published through the Information Bridge: DOE Scientific and Technical Information. "IS-T 1944" Lisa M. Ponton. 12/19/2004. Report is also available in paper and microfiche from NTIS.

Wheat bran is a promising material containing a wide range of useful components, including proteins. In addition, it is produced in significant volumes. Currently, wheat bran is used for the production of energy by combustion and for feed purposes. Gradually, new methods of valorization of this material are being sought. One of the possibilities of using wheat bran is the isolation of proteins, hydrolysis, and separation of selected amino acids. This diploma thesis deals with this issue, it is focused on the recovery of arginine and leucine from a protein isolate. Proteins were extracted from wheat bran by changing the pH. Thanks to the subsequent lyophilization a protein isolate was gained. Prior to hydrolysis of the resulting isolate, a stability test of arginine and leucine amino acid standards was first performed, to which various hydrolysis methods were applied. Acid hydrolysis using a mineralizer, which was applied to the protein isolate, was proved to be the most effective. This was followed by the derivatization of the hydrolysates with OPA and analysis of the resulting hydrolysates by high-performance liquid chromatography with UV-VIS detection. Then, suitable adsorption and desorption conditions were optimized. It was found that the time dependence does not affect the amount of adsorbed material on the sorbent. Therefore, an application time of 15 minutes was chosen. While optimizing the amount of used standard, it was found that the optimal weight was 0.25 g of sorbent. The selected conditions were applied to the protein hydrolyzate. Two fractions were obtained by the separation of selected amino acids due to the change in the pH of the citrate buffer. After the application of this procedure, 0.26 g of arginine and 0.82 g of leucine were obtained from one kilogram after evaporation. From evaporation two, 1.01 g of arginine and 0.25 g of leucine were obtained after evaporation.