Purification of red blood cells by separation and infiltration
A red blood cell and diafiltration technology, applied in the field of red blood cell purification by separation and diafiltration, can solve the problems of reduced yield, variable processing time, cell lysis, etc., and achieve the effect of increasing yield
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Embodiment 1
[0065] Example 1 - Laboratory scale experiments
[0066] Referring to the figure, recover calf whole blood in a container with anticoagulant (EDTA), and at 2,600 rpm (1,200XG), at 4°C, separate the whole blood into the heavy phase (red blood cell component, or cell component ) and the light phase (liquid fraction) in a centrifuge 27 (Beckman J2-21 using a JA-10 centrifuge) for 30 minutes. The starting volume of blood is 200ml. The red blood cell and liquid fractions were separated and the phases were processed in a laboratory-scale cell washing system. Liquid components are processed directly. The red blood cell fraction was directed to the recirculation vessel 28 and diluted to its original volume with isotonic citric acid / salt buffer (sodium citrate hydrate, 6.0 g / L, sodium chloride 8 g / L). The recirculation vessel is maintained at the proper temperature around the recirculation vessel by recirculating the appropriate medium in the recirculation bottle, 30 . Separation ...
Embodiment 2
[0070] Example 2 - Laboratory scale experiment II
[0071] Calf blood was recovered in aseptically processed stainless steel containers containing sodium citrate anticoagulant and batch centrifuged in a second centrifuge type, a CEPA tube-enteric centrifuge (New Brunswick Scientific Corporation, Edison, NJ) Red blood cell component and liquid component. Centrifugation was performed in two feeding configurations. In one configuration, food is pumped in with a peristyle pump, and blood is added in a second configuration using a siphon. The red blood cell fraction obtained from each configuration was then diluted to the original volume of calf blood to produce a blood solution containing isotonic citrate / salt buffer. As a control, citrated whole blood without centrifugation was included as a separate sample. The samples were washed in a bench scale washing apparatus until three retentate volumes of permeate (approximately 600 ml) were obtained. The data are summarized in Ta...
Embodiment 3
[0075] Example 3 - Pilot Scale Experiment
[0076] Blood was pooled from three animals and treated with anticoagulants as described in Example 2. Pooled blood was centrifuged in a Westfalia (Northvale, NJ) SA-1 or SB-7 centrifuge to generate a red blood cell fraction and a liquid fraction, or washed directly as a control. The red blood cell fraction was washed using a pilot-scale washing system.
[0077] Separation of red blood cells was performed through the filter using a pump (Watson-Marlow pump, Wilmington, MA). After passing through the filter, separation of red blood cells was performed by recirculation vessel and diluted with isotonic citrate / salt buffer. The volume of the recirculation vessel is 9.6 liters. The recirculation vessel is maintained at an appropriate temperature by recirculating glycol through recirculation bottles surrounding the recirculation vessel. Separation of the red blood cell fraction / buffer mixture into permeate and retentate was performed ...
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