Lacto
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Posts: 6
Registered: 26-11-2018
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Lactoferrin conversion Apolactoferrin
I’d like your opinion on producing Apolactoferrin (iron free) from Lactoferrin. This is the method I have found.(below)
Is this do-able by a back yard amateur chemist? Is it realistic for the scale of 3 kilos lactoferrin for conversion? Other methodologies?
Can anyone recommend a commercial operation that might do it?
Thanks.
Lactoferrin preparation
Apo-bLf (iron free) was prepared from commercial grade pure, endotoxin (LPS) free, native bLf. Briefly, 80 mg/ml native bLf was dissolved in Milli-Q
water and iron released by reducing pH to 2.06. The bLf solution was then dialysed in 50 kDa molecular weight cut-off dialysis tubing against 0.1 M
citric acid for 48 h and pH adjusted back to 8.0. Fe-bLf (iron-saturated) was prepared by the addition of ferric nitriloacetate (Fe-NTA) to Apo-bLf
drop wise until the solution reached a deep red colour indication iron saturation. The Fe-bLf solution was then dialysed against Milli-Q water for 48
h. Protein estimation was performed using the Coomassie Plus (Bradford) Protein Assay (Thermo Scientific) and purification was confirmed via SDS-PAGE
(results not shown).
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Metacelsus
International Hazard
Posts: 2539
Registered: 26-12-2012
Location: Boston, MA
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Mood: Double, double, toil and trouble
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Getting the proper dialysis tubing on such a large scale might be an issue. Otherwise it seems OK. But how are you going to get the 3 kg lactoferrin
to start with? That's a very large scale for a protein.
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