isolation of protein using preparative HPLC

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srujana
srujana's picture
isolation of protein using preparative HPLC

Dear Sir/Madam,

I am interested in isolating lactoferrin protein(77kd) from whey and whose concentration would be between 20-60mg/ml.I would like to know which preparative column is more suitable for this work.I would also like to know whether the instrument used for Analytical HPLC can also be used for Preparative HPLC.Please help me out as iam very new to this field

Srujana

skarthikumar
skarthikumar's picture
The analytical HPLC can't be

The analytical HPLC can't be used for seperation of components. It can only give the chromotogram where you can see the purity of your compound.

Dr. Analytical
Dr. Analytical's picture
Srujana:

Srujana:
Usually, you will develop a separation method for your components on an analytical scale column, and then "scale-up" the separation with a column of the same type with a larger diameter.

There are two problems with using an analytical scale instrument:
1. The maximum flow rate may not be large enough.  5 mL/min is a typical limit, but large columns may require 10 - 20 mL/min.

2. The injectors with usually have a small volume limit (100 uL or less), so you would have to make many injections.