Mobilization of focused protein zones by ion introduction

Details Mobilization of focused protein zones by ion introduction Mobilization of focused protein zones by ion introduction

1988-10-18

1990-03-27

Niebling, John F.

Chemistry: electrical and wave energy

Processes and products

Electrostatic field or electrical discharge

2041832, G01N 2726, B01D 5702

Patent

active

049118080

ABSTRACT:
Zones of ampholytic substances which have been isoelectrically focused in a separation medium are mobilized for analytical or preparative purposes by either introducing anions other than hydroxyl ions to the separation medium toward the catholyte end or adding cations other than protons to the separation medium toward the anolyte end to produce a pH shift in the separation medium, thus restoring a charge to the focused ampholytes so that they will mobilize under the influence of a voltage applied across the medium. In preferred embodiments, the anion or cation introduction is achieved by supplementing the catholyte or anolyte, respectfully, with a strong electrolyte and permitting the cations or anions to migrate into the separation medium under the influence of the electric field. Detection is achieved by permitting the focused zones to migrate past a fixed point in the separation medium. Alternatively, detection of the zones outside of the separation medium and/or recovery of the individual proteins themselves may be achieved in the same way provided the receiving electrolyte has an appropriate pH which will permit migration of the zones to continue into the electrolyte itself.

REFERENCES:
McCormick, A. et al., "Selective Elution of Zones from Preparative Isoelectric Focusing Gels by Ampholytes and Buffers" Analytical Biochemistry 75, 314-324 (1976).
McCormick, A. et al., "Preparative Separation of Hemoglobins A and S by Gel Electrofocusing, Using Selective Zone Elution by Gel Transposition between Suitable Anolytes and Catholytes" Analytical Biochemistry 85, 209-218 (1978).

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