This document introduces a method for separating immunoglobulins using size-exclusion chromatography. The method uses a high-performance ZORBAX particle that is extremely rugged, even under high back-pressures. This characteristic allows higher flow rates and shorter run times for high-throughput experimentation. The method can also reduce the injection volume of an antibody sample without loss of sensitivity, when using smaller-diameter columns.
This document describes a method for separating IgM and IgG antibodies using size-exclusion chromatography. The procedure utilizes a ZORBAX GF-250 column with a mobile phase of 200 mM Sodium Phosphate (pH 7) containing 0.01% Azide. The separation is achieved in less than 4 minutes, highlighting the ruggedness and high flow rate capabilities of the ZORBAX particles compared to polymer-based materials.
| Column | ZORBAX GF-250 (4.6 x 250 mm) |
|---|---|
| Mobile Phase | 200 mM Sodium Phosphate (pH 7), 0.01% Azide |
| Injection Volume | 2.5 pL (1 mg/mL) |
| Flow Rate | 0.94 mL/min |
| Detection | UV (230 nm) |
| Run Time | <4 min |
The application uses a ZORBAX GF-250 column with dimensions of 4.6 x 250 mm.
The procedure specifies a flow rate of 0.94 mL/min for the separation.
The text states that run times are shorter than 4 minutes for high-throughput experimentation.