Apparatus and process for the large scale separation and retrieval of proteins from cellular extracts

A pretreatment device for separating a mass of cellular extract into a crude mixture of proteins amenable to further purification, said device comprising:

• A. a receptacle for storing a conductive buffer solution, said receptacle having a generally horizontal floor;
• B. first and second electrodes positioned in said receptacle;
• and
• C. a plurality of cassettes disposed in said receptacle between said first and second electrodes and extending upwardly from said floor, each of said cassettes including
• (1) first and second semipermeable membranes, and
• (2) a layer of a conductive gel matrix sandwiched between said first and second membranes.

A system for separating and retrieving highly purified proteins from a crude cellular extract, comprising:

• A. a pretreatment device for separating a mass of said crude cellular extract into a crude mixture of proteins amenable to further purification, said device including
• (1) a receptacle for storing a conductive buffer solution, said receptacle having a generally horizontal floor;
• (2) first and second electrodes positioned in said receptacle;
• and
• (3) a plurality of cassettes disposed in said receptacle between said first and second electrodes and extending upwardly from said floor, each of said cassettes including
• (A) first and second semiperlneable membranes, and
• (B) a layer of a conductive gel matrix sandwiched between said first and second membranes;
• and
• B. an electrophoretic device which receives said crude mixture of proteins from said pretreatment device and which separates and retrieves said highly purified proteins from said mixture.

A method of separating and retrieving highly purified proteins from a crude cellular extract, comprising:

• A. separating a mass of said crude cellular extract into a crude mixture of proteins amenable to further purification, said separation step including
• (1) inserting said mass into a receptacle between first and second ends extending from a generally horizontal floor, a plurality of cassettes being disposed in said receptacle between said first and second ends and extending upwardly from said floor, a conductive buffer solution being disposed in said receptacle between said cassettes, then
• (2)conducting an electric current through said buffer solution to cause said mass to migrate from said first end of said receptacle to said second end,
• (3) impeding the passage of groups of larger proteins through each of successive ones of said cassettes while permitting groups of smaller proteins to pass therethrough more rapidly, thus producing said crude mixture, and then
• (4) removing said crude mixture from said receptacle;
• and then
• B. separating and retrieving highly purified enzymes from said crude mixture through continuous vertical gel electrophoresis.