Gels Mimicking Antibodies in Their Selective Recognition of Proteins and Its Potential Use for Protein Crystallization - Molecules: Nucleation, Aggregation and Crystallization - page 14

L of a sample solu-
tion of hemoglobin (10 mg/mL) and ribonuclease (3 mg/mL) was applied.
The column was then washed with 0.01 M sodium phosphate, pH 7.0, and
the eluate was collected and analyzed by cation-exchange chromatogra-
phy at 220 nm.
The results are shown in the chromatogram traces of Fig. 1. Figure l(a)
is a chromatogram of the sample itself prior to passage through any
gel column (“Hb” designating hemoglobin and “R” designating ribo-
nuclease). Figure l(b) is a chromatogram of the fraction collected from a
blank column (prepared in the absence of ribonuclease). Figure l(c) is a
chromatogram of the fraction collected from the column prepared in the
presence of ribonuclease. The ribonuclease peak is present in Figs. l(a)
and l(b), but absent in Fig. l(c), indicating that ribonuclease was adsorbed
only by the column prepared in the presence of ribonuclease.

3 mL of 0.01 M sodium phosphate, pH 7.0. About 50 µ

Fig. 1. Selective recognition of ribonuclease. Analysis in a cation exchanger of (a) the
sample itself [consisting of hemoglobin (Hb) and ribonuclease (R)] prior to passage
through any gel column, (b) the fraction collected from a blank column (prepared in the
absence of ribonuclease) and (c) the fraction collected from the column prepared in the
presence of ribonuclease. The ribonuclease peak is present in (a) and (b), but absent in
(c), indicating that ribonuclease had been specifically adsorbed only onto the gel column
prepared in the presence of ribonuclease.