Abstract
Enzymatic conversion of type B to O erythrocytes with Coffea (coffee bean) α-D-galactosidase was first described by Harpaz and Flowers and subsequently adopted by others (1,2). An enzyme-linked immunosorbent assay (ELISA) and soluble oligosaccharide substrates were used to study deantigenation of B erythrocytes with the Coffea enzyme. For the ELISA, microtiter wells were coated with B membranes and treated with enzyme under a variety of conditions, probed with primary IgM monoclonal anti-B followed by secondary anti-murine μ-chain specific alkaline phosphatase conjugate, then developed with substrate. This technique has allowed the rapid determination of enzymatic activity over a broad range of conditions; the purpose being to determine parameters for efficiently enhancing enzyme activity. Solid phase activity then compared to activity against soluble oligosaccharide substrates. We have determined that, under the conditions tested, only moderate increases in enzyme activity against the B epitope can be achieved by modifying reaction conditions with the native Coffea enzyme.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 489-502 |
| Number of pages | 14 |
| Journal | Artificial Cells, Blood Substitutes, and Immobilization Biotechnology |
| Volume | 24 |
| Issue number | 5 |
| State | Published - Sep 1996 |
| Externally published | Yes |
Fingerprint
ASJC Scopus subject areas
- Biomedical Engineering
- Hematology
- Biotechnology
- Biomaterials
Cite this
Characterization of Coffea canephora α-D-galactosidase blood group B activity. / Phillips, Roy; Smith, Daniel.
In: Artificial Cells, Blood Substitutes, and Immobilization Biotechnology, Vol. 24, No. 5, 09.1996, p. 489-502.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Characterization of Coffea canephora α-D-galactosidase blood group B activity
AU - Phillips, Roy
AU - Smith, Daniel
PY - 1996/9
Y1 - 1996/9
N2 - Enzymatic conversion of type B to O erythrocytes with Coffea (coffee bean) α-D-galactosidase was first described by Harpaz and Flowers and subsequently adopted by others (1,2). An enzyme-linked immunosorbent assay (ELISA) and soluble oligosaccharide substrates were used to study deantigenation of B erythrocytes with the Coffea enzyme. For the ELISA, microtiter wells were coated with B membranes and treated with enzyme under a variety of conditions, probed with primary IgM monoclonal anti-B followed by secondary anti-murine μ-chain specific alkaline phosphatase conjugate, then developed with substrate. This technique has allowed the rapid determination of enzymatic activity over a broad range of conditions; the purpose being to determine parameters for efficiently enhancing enzyme activity. Solid phase activity then compared to activity against soluble oligosaccharide substrates. We have determined that, under the conditions tested, only moderate increases in enzyme activity against the B epitope can be achieved by modifying reaction conditions with the native Coffea enzyme.
AB - Enzymatic conversion of type B to O erythrocytes with Coffea (coffee bean) α-D-galactosidase was first described by Harpaz and Flowers and subsequently adopted by others (1,2). An enzyme-linked immunosorbent assay (ELISA) and soluble oligosaccharide substrates were used to study deantigenation of B erythrocytes with the Coffea enzyme. For the ELISA, microtiter wells were coated with B membranes and treated with enzyme under a variety of conditions, probed with primary IgM monoclonal anti-B followed by secondary anti-murine μ-chain specific alkaline phosphatase conjugate, then developed with substrate. This technique has allowed the rapid determination of enzymatic activity over a broad range of conditions; the purpose being to determine parameters for efficiently enhancing enzyme activity. Solid phase activity then compared to activity against soluble oligosaccharide substrates. We have determined that, under the conditions tested, only moderate increases in enzyme activity against the B epitope can be achieved by modifying reaction conditions with the native Coffea enzyme.
UR - http://www.scopus.com/inward/record.url?scp=0030245549&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0030245549&partnerID=8YFLogxK
M3 - Article
C2 - 8879423
AN - SCOPUS:0030245549
VL - 24
SP - 489
EP - 502
JO - Artificial Cells, Nanomedicine and Biotechnology
JF - Artificial Cells, Nanomedicine and Biotechnology
SN - 2169-1401
IS - 5
ER -