"Inside-Out" PEGylation of Bovine ß-Cross-Linked Hemoglobin.
Artif Organs
; 41(4): 351-358, 2017 Apr.
Article
en En
| MEDLINE
| ID: mdl-28321886
ABSTRACT
The development of a blood substitute is urgent due to blood shortages and potential communicable diseases. A novel method, inside-out PEGylation, has been used here to conjugate a multiarm maleimide-PEG (Mal-PEG) to ß-cross-linked (ßXL-Hb) hemoglobin (Hb) tetramers through the Cys ß93 residues. This method produces a polymer with a single PEG backbone that is surrounded by multiple proteins, rather than coating a single protein with multiple PEG chains. Electrophoresis under denaturing conditions showed a large molecular weight species. Gel filtration chromatography and analytical ultracentrifugation determined the most prevalent species had three ßXL-Hb to one Mal-PEG. Thermal denaturation studies showed that the cross-linked and PEGylated species were more stable than native Hb. Cross-linking under oxy-conditions produced a high oxygen affinity Hb species (P50 = 9.18 Torr), but the oxygen affinity was not significantly altered by PEGylation (P50 = 9.67 Torr). Inside-out PEGylation can be used to produce a hemoglobin-based oxygen carrier and potentially for other multiprotein complexes.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Polietilenglicoles
/
Sustitutos Sanguíneos
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Hemoglobinas
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Reactivos de Enlaces Cruzados
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Composición de Medicamentos
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Maleimidas
Límite:
Animals
Idioma:
En
Revista:
Artif Organs
Año:
2017
Tipo del documento:
Article
País de afiliación:
Israel