Purification of exopolygalacturonase by affinity chromatography on Concanavalin A — Bead cellulose

1996 ◽  
Vol 10 (5) ◽  
pp. 363-366
Author(s):  
E. Stratilová ◽  
D. Mislovičová ◽  
M. Dzúrová
Keyword(s):  
Affinity Chromatography ◽  
Concanavalin A ◽  
Bead Cellulose

Affinity chromatography used in distinguishing alpha-fetoprotein in serum from patients with tumors of hepatic parenchyma and of germ cells.

1984 ◽  
Vol 30 (7) ◽  
pp. 1257-1258 ◽  
Author(s):  
P K Buamah ◽  
C Cornell ◽  
A W Skillen
Keyword(s):  
Germ Cell ◽  
Affinity Chromatography ◽  
Germ Cells ◽  
Concanavalin A ◽  
Primary Liver Cancer ◽  
Germ Cell Tumors ◽  
Alpha Fetoprotein ◽  
Hepatic Parenchyma ◽  
Liver Cancers ◽  
Serum Alpha Fetoprotein

Abstract We used affinity chromatography on concanavalin A Sepharose to study the serum alpha-fetoprotein of 10 patients with histologically proven germ-cell tumors and 12 patients with primary liver cancer. Less than 50% of the fetoprotein from germ-cell tumors bound to concanavalin A, as compared with more than 80% of the alpha-fetoprotein from primary liver cancers.

scholarly journals Serological reactivity of different antigenic preparations of Leishmania (Leishmania) amazonensis and the Leishmania braziliensis complex

2008 ◽  
Vol 41 (2) ◽  
pp. 135-141 ◽  
Author(s):  
Adriano Gomes-Silva ◽  
Maria Aparecida Souza ◽  
Sandra Regina Afonso-Cardoso ◽  
Lívia Resende Andrade ◽  
Reynaldo Dietze ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Affinity Chromatography ◽  
Healthy Volunteers ◽  
Concanavalin A ◽  
Leishmania Amazonensis ◽  
Leishmania Braziliensis ◽  
Elisa Assay ◽  
Serum Samples ◽  
Sensitivity Range ◽  
Serological Reactivity

Total antigen from Leishmania (Leishmania) amazonensis and isolates from the Leishmania braziliensis complex, along with their respective antigenic fractions obtained by affinity chromatography on concanavalin-A-Sepharose and jacalin-agarose columns evaluated using immunoenzymatic ELISA assay. For this, serum samples from 229 patients were used, grouped as American tegmental leishmaniasis (nº=58), visceral leishmaniasis (nº=28), Chagas disease (nº=49), malaria (nº=32), tuberculosis (nº=13) and healthy volunteers (nº=49). Samples from American tegmentary leishmaniasis showed higher reactivity with antigens isolated from the Leishmania braziliensis complex than with antigens from Leishmania amazonensis (p<0.001). ELISA assays showed a sensitivity range from 60% to 95% with antigens isolated from the Leishmania braziliensis complex. There was marked nonspecific reactivity among serum samples with the use of antigenic fractions binding with concanavalin-A and jacalin from both Leishmania complexes, in comparison with other antigens (p<0.001). The results presented in this study suggest that the use of homologous antigens increases the efficiency of anti-Leishmania immunoglobulin detection, which may be very valuable for diagnostic purposes.

scholarly journals Characterization of human liver α-D-mannosidase purified by affinity chromatography

1976 ◽  
Vol 153 (3) ◽  
pp. 579-587 ◽  
Author(s):  
N C Phillips ◽  
D Robinson ◽  
B G Winchester
Keyword(s):  
Affinity Chromatography ◽  
Concanavalin A ◽  
Human Liver ◽  
Early Stage ◽  
Deae Cellulose ◽  
Enzymic Activity ◽  
Exchange Chromatography ◽  
Lysosomal Storage ◽  
Sepharose 4B

Human liver acidic α-D-mannosidase was purified 1400-fold by a relatively short procedure incorporating chromatography on concanavalin A-Sepharose and affinity chromatography on Sepharose 4B-epsilon-aminohexanoylmannosylamine. In contrast with the acidic enzymic activity the neutral α-mannosidase did not bind to the concanavalin A-Sepharose so the two types of α-mannosidase could be separated at an early stage in the purification. The only significant glycosidase contaminant after affinity chromatography on the mannosylamine ligand was α-L-fucosidase, which was selectively removed by affinity chromatography on the corresponding fucosylamine ligand. The final preparation was free of other glycosidase activities. The pI of the purified enzyme was increased from 6.0 to 6.45 on treatment with neuraminidase. Although the pI and the mol.wt. (220 000) suggested that α-mannosidase A had been purified selectively, ion-exchange chromatography on DEAE-cellulose indicated that the preparation consisted predominantly of α-mannosidase B. This discrepancy is discussed in relation to the basis of the multiple forms of human α-mannosidase. The purified enzyme completely removed the α-linked non-reducing terminal mannose from a trisaccharide isolated from the urine of a patient with mannosidosis. A comparison of the activity of the pure enzyme towards the natural substrate and synthetic substrates suggests that the same enzymic activity is responsible for hydrolysing all the substrates. These results validate the use of synthetic substrates for determining the mannosidosis genotype. They are also further evidence that mannosidosis is a lysosomal storage disease resulting from a deficiency of acidic α-mannosidase.

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