Size enhanced hemoglobins: surface decoration and...

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...

Reexamination Certificate

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C530S402000, C514S002600

Reexamination Certificate

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11295279

ABSTRACT:
Novel modified hemoglobins comprising polyalkylene glycols and novel methods for making those hemoglobins are provided. One group of modified hemoglobins comprise polyalkylene glycols bonded to the hemoglobin with an amide linkage at Glu-43(β). Additional polyalkylene glycols can also be bonded to the Glu-22(β) and/or the Asp-47(β). These hemoglobins are made by a novel amidation procedure. A second group of modified hemoglobins comprise a polyalkylene glycol covalently bonded to the hemoglobin at the α-amino of a Val-1(β) or a Val-1(α). Additional polyalkylene glycols can optionally be covalently bonded to a limited number of ε-amino groups. This second group of hemoglobins is made using a novel reductive alkylation procedure. A third group of modified hemoglobins comprise a polyalkylene glycol bonded to a thiol group of the hemoglobin through a phenylsuccinimido linkage, wherein no polyalkylene glycol is bonded to a Cys-93(β). This third group of modified hemoglobins is made by an improvement in a hemoglobin-polyalkylene linkage procedure utilizing thiolation-mediated maleimide chemistry.

REFERENCES:
patent: 4412989 (1983-11-01), Iwashita et al.
Rao et al. ‘Amidation of Basic Carboxyl Groups of Hemoglobin,’ Methods in Enzymology. vol. 231. pp. 246-267. 1994.

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