An aqueous 0.1% formic acid solution was used as phase A and a solution of 0.1% formic acid and 84% acetonitrile was used as phase B. The peptides were separated with a linear gradient of 4% ∼50%mobile phase B over 2 hours at a flow rate of 1300 nl/min.
Treatment was carried out for 3 hours at a flow rate of 80 100 ml/minute, and DHP-PMX was performed twice within 24 hours.
A double-lumen catheter was placed via a femoral or internal jugular vein, and PMX-DHP was administered for 6 or 12 hours (usually 12 hours) at a flow rate of 80 100 mL/minute.
A solution of HuLF in 0.1 M carbonate buffer, pH 9.5, was re-circulated over the column for 16 hours at a flow rate of 0.5 ml/min at 4°C.
After coupling, the column was washed with 0.1 M carbonate buffer, pH 9.5, until all non-bound HuLF had been removed, followed by re-circulation of 25 ml 0.1 M ethanolamine in 0.1 M carbonate buffer, pH 9.5, for 3 hours at a flow rate of 1 ml/min to block the remaining active epoxy groups.
DHP-PMX was performed for two hours at a blood flow rate of 50 to 150 ml/minute, once [ 15, 21, 28, 30- 32, 34] or a maximum of two [ 8, 9, 11- 16, 16- 20, 22- 29, 29, 33, 35] or three [ 38] times, depending on the clinical response of the patient.
Upon reaching the growth temperature (TG) the flow of NH3was maintained at 250 sccm for a further 60 min after which the tube was allowed to cool down over at least an hour, in a flow of 50 sccm NH3.
Sucrose solution at 5% was supplied for 6 minutes every 8 hours with a flow rate of 15 mL/hr.
Here the tides are very high and the current strong, reaching about 6 knots, or 7 miles per hour, at peak flows four times a day.
The solutions were delivered at a flow rate of 0.5ul per hour for 2 weeks.
