Engineering of the LukS-PV and LukF-PV subunits of Staphylococcus aureusPanton-Valentine leukocidin for Diagnostic and Therapeutic Applications

Table 3 Schema of the Ni⁺⁺-NTA fast protein liquid chromatography programme^a,b

Step	Volume (mL)^c	Solution^d
01	00 - 20	D^e
02	20 - 40	C^f
03	40 - 60	A
04	60 – 90	E = Protein
05	90 – 115	A
06	115 – 130	Mix 5% B
→ 120 Collect 2 mL
07	130 - 180	Gradient of 5 to 100% B
08	180 - 200	B
→ Divert to waste
End of method, washing up of the machine, and log off

→ No necessary information. Proceed to the shown step.
^a The flow-rate was 1.0 mL throughout the entire run.
^b The entire programme was 200 mL.
^c Increase on the tabulated volumes were always allowed by pushing the Hold (H) button on the machine. This accounts for the ‘H’ on the BioLogic LP output during the run.
^d All solutions were filtered and degassed.
^e Solution D is sterile distilled water.
^f Solution C is the charge buffer.

ISSN: 1472-6750