Characterization of Coffea canephora α-D-galactosidase blood group B activity

Roy Phillips, Daniel Smith

Research output: Contribution to journalArticle

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 languageEnglish (US)
Pages (from-to)489-502
Number of pages14
JournalArtificial Cells, Blood Substitutes, and Immobilization Biotechnology
Volume24
Issue number5
StatePublished - Sep 1996
Externally publishedYes

Fingerprint

Galactosidases
Coffee
Blood Group Antigens
Blood
Enzymes
Oligosaccharides
Immunosorbents
Enzyme activity
Assays
Substrates
Erythrocytes
Enzyme-Linked Immunosorbent Assay
Epitopes
Phosphatases
Alkaline Phosphatase
Immunoglobulin M
Rho(D) antigen
Membranes

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 journalArticle

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