Transfusions to group O subjects of 2 units of red cells enzymatically converted from group B to group O

Lenny, Leslie L.; Hurst, Rosa; Goldstein, Jack; Galbraith, R.A.

doi:10.1046/j.1537-2995.1994.34394196617.x

Cited by 56 publications

(24 citation statements)

References 11 publications

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“…Isoform III resembled isoform IA in having the same N-terminal amino acid sequence, but it differed from IA and IB in having a more acidic pH optimum, greater thermal stability, greater sensitivity to metal ions and alkylating agents, and anomalous retardation during gel filtration on Sephacryl S-300. Anomalous retardation of ␤-GlcNAc'ase on derivitized dextran gels has also been observed by us 2 and by others (66 -68); it has been ascribed to hydrophobic interactions as well as to charge effects (69).…”

Section: Discussionsupporting

confidence: 66%

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Purification and Characterization of Blood Group A-degrading Isoforms of α-N-Acetylgalactosaminidase from Ruminococcus torques Strain IX-70

Hoskins

Boulding

Larson

1997

Journal of Biological Chemistry

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To cleave blood group A immunodeterminants from erythrocytes (Hoskins, L. C., Larson, G., and Naff, G. B. (1995) Transfusion 35, 813-821), we purified and characterized ␣-N-acetylgalactosaminidase (EC 3.2.1.49) activity from culture supernatants of the human fecal bacterium Ruminococcus torques strain IX-70. Three isoforms separated during hydrophobic interaction chromatography. Hydroxyapatite chromatography further resolved the most hydrophilic, isoform I, into isoforms IA and IB. The most hydrophobic, isoform III, differed from IA and IB by a more acidic pH optimum, greater heat resistance, greater sensitivity to alkylating agents, and anomalous retardation during gel filtration chromatography. Isoform IB differed from IA and III in N-terminal amino acid sequence and in sensitivity to EDTA inhibition. Each cleaved nonreducing ␣(133)-N-acetylgalactosamine residues from human blood group A and AB mucin glycoproteins, Forssman hapten, and blood group A lacto series glycolipids. The apparent molecular mass of denatured isoform subunits of IA, IB, and III-PII (158, 173, and 201 kDa, respectively) bore no integer relationship to the apparent molecular mass of the native isoforms (265, 417, and 530 kDa), but the latter bore a ratio of 1.96:3.09:3.93 to the weight-average apparent molecular mass of native IA (135 kDa), suggesting that the isoforms are multimers of a 135-kDa sequence. Isoforms IA and III-PII had an identical N-terminal amino acid sequence which showed homologies to the N-terminal sequence of sialidases produced by Bacteroides fragilis SBT3182, another commensal enteric bacterium.

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Section: Discussionsupporting

confidence: 66%

“…Lenny and associates (2)(3)(4) have demonstrated that blood group B red cells whose B immunodeterminants were removed by coffee bean ␣-galactosidase survive normally in blood group A and O recipients.…”

mentioning

confidence: 99%

Purification and Characterization of Blood Group A-degrading Isoforms of α-N-Acetylgalactosaminidase from Ruminococcus torques Strain IX-70

Hoskins

Boulding

Larson

1997

Journal of Biological Chemistry

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“…Furthermore, there was a commercial application of the enzyme as a supplement in animal feed to increase animal meat production. Finally, rice α-galactosidase may be useful for blood conversion purpose (Harpaz et al 1977;Lenny et al 1994;Zarnitz and Kabat 1960;Zhu et al 1995) since its ability to convert blood type is comparable with that of the taro enzyme (Chien 1991). In fact, we were the first to show that the rice α-galactosidase was capable to convert B red blood cells into O red cells.…”

Section: Discussionmentioning

confidence: 91%

Cloning, Expression, and Characterization of Rice α-Galactosidase

2008

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We have successfully cloned an α-galactosidase gene from a rice cDNA library and transformed it into Escherichia coli BL21. It was subsequently cloned to the pPIC9K vector and expressed in Pichia pastoris. A selected clone was found to result in high production yield of the galactosidase enzyme. The secreted enzyme was purified, and it revealed as a major protein band on an SDS-PAGE gel. The optimal pH value, enzyme stabilities, and substrate specificity were studied. The enzyme specificity toward the terminal α1→6, 1→4, and 1→3 linked galactosyl residue from various substrates was investigated. By determining the Michelis constant (Km) of the enzyme for melibiose, raffinose, and stachyose, our results showed that melibiose was hydrolyzed faster than raffinose, whereas the published data reported a reversed sequence, raffinose > melibiose. The enzyme also showed the ability of converting B red blood cells into O red cells. The objective of this work is to develop the Pichia system to produce a large quantity of enzyme for blood cell conversion for transfusion.

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“…Despite this, coffee-bean-derived enzyme was successfully used to complete phase I (Lenny et al, 1991(Lenny et al, , 1994(Lenny et al, , 1995 and phase II clinical trials in which B-ECO RBCs were shown to be safe and functional, providing RBC survival and recovery values equal to ABO-compatible control transfusions in patients during a randomized cross-over study (Kruskall et al, 2000).…”

Section: The Eco Process and The Enzymesmentioning

confidence: 99%

“…Clinical phase I studies were conducted using B-ECO RBCs given to group A and O healthy volunteers and the initial small infusions were escalated to full single RBC units (Lenny et al, 1991), multiple units and finally repeated transfusions (Lenny et al, 1994(Lenny et al, , 1995. A successful phase II cross-over clinical trial in patients was reported in 2000 (Kruskall et al, 2000).…”

Section: Clinical Experience With Eco Rbcsmentioning

confidence: 99%

Modifying the red cell surface: towards an ABO‐universal blood supply

Olsson¹,

Clausen²

2007

Br J Haematol

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SummaryEliminating the risk for ABO-incompatible transfusion errors and simplifying logistics by creating a universal blood inventory is a challenging idea. Goldstein and co-workers pioneered the field of enzymatic conversion of blood group A and B red blood cells (RBCs) to O (ECO). Using a-galactosidase from coffee beans to produce B-ECO RBCs, proof of principle for this revolutionary concept was achieved in clinical trials. However, because this enzyme has poor kinetic properties and low pH optimum the process was not economically viable. Conversion of group A RBCs was only achieved with the weak A 2 subgroup with related enzymes having acidic pH optima. More recently, the identification of entirely new families of bacterial exoglycosidases with remarkably improved kinetic properties for cleaving A and B antigens has reinvigorated the field. Enzymatic conversion of groups A, B and AB RBCs with these novel enzymes resulting in ECO RBCs typing as O can now be achieved with low enzyme protein consumption, short incubation times and at neutral pH. Presently, clinical trials evaluating safety and efficacy of ECO RBCs are ongoing. Here, we review the status of the ECO technology, its impact and potential for introduction into clinical component preparation laboratories.

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Transfusions to group O subjects of 2 units of red cells enzymatically converted from group B to group O

Abstract: These results extend those observed in our earlier 1-unit transfusion studies and suggest that ECO RBCs pose little risk and will be useful in transfusion medicine.

Cited by 56 publications

References 11 publications

Purification and Characterization of Blood Group A-degrading Isoforms of α-N-Acetylgalactosaminidase from Ruminococcus torques Strain IX-70

Purification and Characterization of Blood Group A-degrading Isoforms of α-N-Acetylgalactosaminidase from Ruminococcus torques Strain IX-70

Cloning, Expression, and Characterization of Rice α-Galactosidase

Modifying the red cell surface: towards an ABO‐universal blood supply

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