One Step Protein Purification -
works directly in undiluted cell Lysate
- More efficient mixing than any other bead technology
- Particle size can be optimized for solutions of different viscosities
- Reduce need for other types of purification, e.g.
- - Affinity matrix columns
- - Batch processes
- Reduce steps and costly reagents
- Increase recovery
- Can reduce time and cost of purification
Capture of Specific Protein from Undiluted Whole Blood
It is often difficult to isolate a specific protein from blood. Because of the complexity and viscosity of whole blood most detection and isolation procedures require the removal of cells and dilution of the sample. CAPs magnetic particles have demonstrated efficient mixing, superior selectivity and rapid magnetic separation of specific proteins from undiluted whole blood and whole serum. Two experiments were designed to demonstrate this ability.
In the first experiment, mouse peroxidase-anti-peroxidase (mPAP) complexes were spiked at concentrations of 0 ng/ml to 100 ng/ml into undiluted human AB whole blood. CAPs coupled with goat-anti-mouse antibodies (GAM-CAPs) were incubated in the blood containing the mPAP complexes for 10 minutes, a magnetic field was applied to retain the CAP protein complex, and the blood was decanted. After washing, tetramethyl benzidine (TMB) was incubated for several minutes to develop color. Absorbance at A450 (y axis) demonstrates that the complexes were effective captured and removed from undiluted whole blood (Figure 1).
In a second experiment, streptavidin-coupled CAPs (SA-CAPs) were used to remove biotinylated horseradish peroxidase (HRP) from undiluted whole human AB serum. As in the first experiment SA-CAPs were incubated for 10 minutes in undiluted whole serum spiked with varying concentrations of biotinylated HRP ranging from 0 ng/ml to 50 ng/ml. The mixture was applied to a magnetic field to remove the captured HRP protein. The HRP color was developed with TMB as above. Absorbance at A450 demonstrates that the SA-CAPs were effective in capturing the biotinylated HRP from undiluted whole serum. Preincubation with100 ng mouse IgG1 blocked the specific capture of the biotinylated HRP (Figure 2). The linearity of the blocked data indicate effective competition for the particle binding sites.
