OraChrom, Inc.

  
       

 

 

Home
Up

 

APPLICATION NOTE

AN122401-20

Metal Chelate Liquid Chromatography on Hard Gel Gigaporous Polymeric Media: Comparison with Soft Gel.

Metal chelate Liquid chromatography includes three major steps of:
a- metal loading,
b- protein adsorption and
c- gradient or stepwise elution of the adsorbed proteins.
 

 

 

Due to flow and pressure restrictions, a typical soft gel metal chelate media would require hours of equilibration to be ready for an actual run of 20 to 30 minutes.
 


The same separation can be performed on a hard gel gigaporous polymeric STYROSä MC-IDA or MC-TED in much shorter time without compromising the resolution.

 

Table 1. Operating parameters for chromatograms 2 and 3. 

HPLC System.

HP 1100

Column

As indicated

Mobile Phase

A: 20 mM Sodium Phosphate, 1 M NaCl, pH = 7.5
B: 20 mM Sodium Phosphate, 1 M NH4Cl, pH = 7.5

Flow rate

2.5 ml/min (900 cm/hr)

Gradient

0 to 100% B in 12 Column Volume

Temperature

30°C

Detection

280 nm

Injection volume

5 ml

Sample:
(8:10 mg/ml each respectively).

1: Ribonuclease A (bovine)
2: apo-Transferrin (human)
(dissolved in 50 % buffer A) Proteins are assessed by the supplier to be 99% pure.

A typical STYROSä MC-IDA column loaded with Cu++ can be used in as many as 50 separation cycles before it requires any regeneration.

[ Home ] [ Up ]

Copyright © 1997-2009 OraChrom, Inc.