DEAE Affie Gel Blue From Bio-Rad

January 08, 2007

DEAE Affie Gel Blue From Bio-Rad
Bio-Rad’s DEAE Affie Gel Blue is a chromatography solid phase that separates protein biomolecules based upon bifunctional reactivity: i) hydrophobic and hydrophilic interactions, ii) charge functional group interactions which separate the molecules based upon charge-charge interactions as dictated by the pH of the mobile phase and the pI of the protein at that pH. The solid phase was specifically designed for the sepearation of transferrin, albumin and immunoglobulin, as well as other proteins within the blood serum. It allows for the purification of immunoglobins in a short amount of time with little effort, saving both time and reagent cost.

While I worked at the University of Pittsburgh, I was working with a team of individuals who were studying AIDS and the course of the disease through a defined population of men. It was part of a multi-centered epidemiology study which took place in a number of cities across the United States. I was one of many people who worked to analyze samples from the study. Our analyses included a multitude of perspectives: i) detection and quantification of anti-HIV antibodies, ii) production of HIV proteins, iii) presence of reverse transcriptase, iv) white blood cells (WBC) counts. Additionally, there were other statistical analyses which looked at secondary and tertiary diseases and related phenomena. I performed all of the assays in the following article: P Gupta et al, “Detection of Human Immunodeficiency Virus by Reverse Transcriptase Assay, Antigen Capture Assay, and Radioimmunoassay,” Journal of Clinical Microbiology, 25(6)1122-1125, 1987.

I purified immunoglobins from HIV+ and HIV- individuals. Bio-Rad’s DEAE Affie Gel Blue method was simple and does not require a lot of additional equipment. The DEAE Affie Gel Blue solid phase support can be reused and recycled for a number of different purifications of IgG. The initial cost for the solid phase is not too expensive and is significantly cheaper than protein A.

In summary, I highly recommend the solid phase because of ease of use, cost and the number of times the column bed can be regenerated.

Former Scientist II
Division Biochemical/Diagnostics
Boehringer Mannheim
  • <<
  • >>
DEAE Affie Gel Blue From Bio-Rad
The Good

The technology is easy to use, does not require a long complex procedure, is reproducible, produces good/excellent yield, and the solid phase can be regenerated multiple uses.

The Bad

Although the solid phase can be regenerated for multiple uses, if you wish to deviate from the recommended buffers because of your particular buffer system, you will not always get the stated yield of IgG recovery. The leaching of the dye off the column bed is probably one of the most significant reasons why the gel does not last for more purifications.

The Bottom Line

With the advent of Protein A and Protein G solid phases as well as other methods for purification of IgG, Bio-Rad’s DEAE Affie Gel Blue is sometimes overlooked. I highly recommend the procedure and the technology.