Apparatus for automatically preparing cell-free proteins and method for preparing proteins using same
Inventors
Park, Han Oh • Kim, Jong Kab • Han, Ji Won • Cho, You Sang • Kim, Min Jung • Kim, Ha Neul • Lee, Yang Won • Kim, Nam Il
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Assignees
Member
BioneerBioneer is a biotechnology company specializing in molecular biology, offering solutions in nucleic acid synthesis, molecular diagnostics, protein production, and laboratory automation. The company's expertise covers oligonucleotide chemistry, gene synthesis, sample preparation, molecular diagnostics, and analytical services such as mass spectrometry and sequencing, serving clinical, research, and industrial sectors globally.
Bioneer is a biotechnology company specializing in molecular biology, offering solutions in nucleic acid synthesis, molecular diagnostics, protein production, and laboratory automation. The company's expertise covers oligonucleotide chemistry, gene synthesis, sample preparation, molecular diagnostics, and analytical services such as mass spectrometry and sequencing, serving clinical, research, and industrial sectors globally.
Publication Number
US-10287542-B2
Publication Date
2019-05-14
Expiration Date
Abstract
An automated cell-free protein production system comprises: a protein expression reaction unit comprising a reaction vessel that includes a plurality of dialysis tubes, each including a dialysis membrane and being open at its top; a reaction temperature control unit configured to heat or cool the reaction vessel; a pipette array comprising a plurality of pipettes and configured to suck or discharge solutions using the pipettes; a pipette array moving unit configured to move the pipette array in an upward and downward direction, a forward and backward direction or a left and right direction so as to move solutions; a protein purification unit including a magnetic field application device; and a multi-well plate mounting unit having mounted therein a multi-well plate kit configured to supply solutions that are used for protein production.
Core Innovation
The invention provides an automated cell-free protein production system with a protein expression reaction unit that includes a reaction vessel of unitary block form. The reaction vessel has an array of hole pairs at a top face, each hole pair open at the top face and extending interiorly into the unitary block form of the reaction vessel. A cylindrical dialysis tube hole is configured to receive a dialysis tube, and a cylindrical small molecule expression solution hole is transversely spaced from the dialysis tube hole and interconnected by a transversely extending channel for fluid communication.
The system automatically and continuously supplies small molecule expression solution to the small molecule expression solution hole in each hole pair whose dialysis tube hole contains a dialysis tube. A pipette array is configured to suck or discharge solutions, and a pipette array moving unit automatically and periodically translates the pipette array in upward and downward, forward and backward, or left and right directions. The translation inserts pipettes into respective small molecule expression solution holes to mix the solution in fluid communication with the cylindrical dialysis membranes of the dialysis tubes in the array of hole pairs.
The automated system further includes a reaction temperature control unit configured to heat or cool the reaction vessel and a protein purification unit including a magnetic field application device. A multi-well plate mounting unit having mounted therein a multi-well plate kit is configured to supply solutions used for protein production. The purification unit uses magnetic beads and magnetic field application to enable protein purification from the magnetic bead suspension, including washing and elution with a protein elution buffer, and the document describes affinity-tag purification via protease cleavage and removal steps to obtain affinity-tag-free pure target protein.
Claims Coverage
The only independent claim identified (clm-00001) covers an automated cell-free protein production system that integrates a unitary block reaction vessel with paired arrays of dialysis tube holes and small molecule expression solution holes, plus automatic pipette-array translation for periodic mixing. It further integrates reaction temperature control, magnetic-field-based protein purification, and a multi-well plate mounting unit supplying solutions for protein production, including magnetic beads-related solutions and small-molecule expression solution.
Unitary block reaction vessel with hole-pair dialysis and small-molecule solution channels
A reaction vessel of unitary block form comprising an array of hole pairs at a top face, including a cylindrical dialysis tube hole configured to receive a dialysis tube and a cylindrical small molecule expression solution hole transversely spaced apart and interconnected in fluid communication by a transversely extending channel, with a top-open cylindrical dialysis membrane interface.
Automatic and continual small-molecule supply into coupled dialysis membranes
Small molecule expression solution is automatically and continually supplied to the small molecule expression solution hole in each hole pair whose dialysis tube hole contains a dialysis tube, so that the small molecule expression solution is in fluid communication with the cylindrical dialysis membranes of the dialysis tubes.
Periodically translated pipette array for solution mixing in fluid communication with dialysis membranes
A pipette array automatically and periodically translated by a pipette array moving unit inserts pipettes into respective small molecule expression solution holes containing small molecule expression solution, to mix the solution therein that is in fluid communication with the cylindrical dialysis membranes in the dialysis tube holes.
Reaction temperature control in the automated cell-free protein production system
A reaction temperature control unit configured to heat or cool the reaction vessel.
Magnetic-field-based protein purification unit
A protein purification unit including a magnetic field application device.
Multi-well plate mounting unit supplying solutions used for protein production
A multi-well plate mounting unit having mounted therein a multi-well plate kit configured to supply solutions that are used for protein production.
Across the identified independent claim, the central inventive coverage is the combined architecture of a unitary block reaction vessel with interconnected hole pairs for dialysis tubes and small molecule expression solution, and an automatically translated pipette array that periodically mixes solutions that remain in fluid communication with dialysis membranes. This is coupled with reaction temperature control, magnetic-field-based purification using magnetic beads, and a multi-well plate kit supplying the solutions used for protein production.
Stated Advantages
Increased expression as shown by experimental outcomes described in the document, including SDS-PAGE and Western blotting results.
Improved scale/compactness relative to a prior dialysis-by-large-volume approach, as stated in the document.
Successful removal of an affinity tag (histidine tag) as described in the document.
Documented Applications
Automated cell-free protein production using dialysis tubes, small-molecule expression solution supply, periodic mixing via a pipette array, and magnetic-field purification with magnetic beads.
Production of affinity-tagged target proteins and affinity-tag-free pure target protein, including affinity-tag removal via protease cleavage followed by protease removal steps as described in the document.
Interested in licensing this patent?