Use of the labscale tangential flow diafiltrator to remove tetrameric hemoglobin from polyhemoglobin, purify hemolysate, and concentrate polyhemoglobin

Artif Cells Blood Substit Immobil Biotechnol. 2009;37(2):61-8. doi: 10.1080/10731190902742455. Epub 2009 Feb 18.

Abstract

The Labscale tangential flow diafiltration system in conjunction with polyethersulfone Biomax filters (both purchased from Millipore Inc.) were used in the following studies: (1) A crucial step in the preparation of polyhemoglobin (polyHb) is the removal of tetrameric hemoglobin (Hb), which can cause adverse side-effects. The efficiency for this depends on the integrity of the 100kDa filters. Those with lower integrity were not effective whereas those with a higher integrity of 0.075ml/min were more effective. A filter with an integrity of 0.075ml/min can reduce the initial 11.2% tetrameric Hb concentration to 1.3% using a flow rate of 0.2ml/min. Higher flow rate of 2.2ml/min was not as effective. (2) Filtration with a 100kDa filter was quick and efficient in separating stroma-free hemoglobin solutions from cellular debris from hemolysate. (3) Both 5kDa and 100kDa filters are efficient in concentrating hemoglobin solutions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood Substitutes / chemistry
  • Blood Substitutes / isolation & purification
  • Blood Substitutes / metabolism
  • Filtration / instrumentation
  • Filtration / methods*
  • Hemoglobins / chemistry
  • Hemoglobins / isolation & purification*
  • Hemoglobins / metabolism
  • Humans
  • Polymers / chemistry
  • Polymers / metabolism*
  • Protein Multimerization
  • Research / instrumentation
  • Research Design
  • Sulfones / chemistry
  • Sulfones / metabolism*

Substances

  • Blood Substitutes
  • Hemoglobins
  • Polymers
  • Sulfones
  • polyhemoglobin
  • polyether sulfone

Grants and funding