Separation of Mycobacterial Soluble Antigens by Isoelectric Focusing

Separation of two hemolysins from Aeromonas hydrophila by isoelectric focusing.

Two hemolytic proteins (hemolysin I and II) with isoelectric points of 5.5 and 4.3 were purified by isoelectric focusing from the culture supernatant fluid of Aeromonas hydrophila. The purified hemolysins were unstable after dialysis and upon heating at 37 C and were inactivated by proteolytic enzymes. The two hemolysins were lethal for mice in a dose of 10 mug (hemolysin I) and 3 mug (hemolysi...

Microfluidic Protein Separation by Free Flow Isoelectric Focusing

Disposable, inexpensive microfluidic devices have the potential to become a robust tool for proteomic research for separating and identifying proteins and protein complexes. In this work, a preparative scale free flow IEF isoelectric focusing (FFIEF) device was designed, investigated, and optimized. A constant electric field of up to 350V/cm was generated across the separation channel upon opti...

Isoelectric focusing.

Separation of soluble Brucella antigens by gel-filtration chromatography.

Soluble precipitating antigens of Brucella suis have been, in various degrees, purified by filtration on Sephadex gels. The most useful gels employed were Sephadex G-150, Sephadex G-200, and Sepharose 4B. Although not all fractions proved to be immunologically pure, some crude molecular-size estimates of most of the 13 soluble antigens of the Brucella cell could be given. In addition, monospeci...

Isoelectric Focusing from Breast Cancer Immune Complexes by Preparative Recovery of Immunologically Reactive Antibodies and Antigens

A new technique for the recovery of immunologically reactive antibodies and putative antigens from breast cancer-associ ated immune complexes was described. Soluble immune com plexes were isolated from extracellular fluids by 2.5% poly ethylene glycol fractionation and affinity chromatography on Protein A:Sepharose CL-4B. The immune complexes bound to immobilized Protein A were then subjected t...