Factory-on-a-chip for production of biologically derived medicines/biopharmaceuticals/biologics/biotherapeutics

Patent metadata

Field	Value
Publication number	US-12410390-B2
Application number	US-202318329643-A
Country	US
Kind code	B2
Filing date	Sep 11, 2023
Priority date	Jun 7, 2017
Publication date	Sep 9, 2025
Grant date	Sep 9, 2025

Abstract

The present invention provides for a fully integrated microfluidic system capable of producing single-dose amounts of biotherapeutics at the point-of-care wherein protein production, purification and product harvest are all integrated as a single microfluidic device which is portable and capable of continuous-flow production of biotherapeutics at the microscale using a cell-free reaction system.

First claim

That which is claimed is:

1. A factory-on-a-chip microfluidic device comprising at least three separate units that can be stacked together to form a single unit, wherein the separate units comprise:
(i) a microfluidic bioreactor unit equipped with a continuous collection channel for synthesizing a crude protein in a reaction within the microfluidic bioreactor unit;
(ii) a microfluidic mixer/de-bubbler unit communicatively connected to the microfluidic bioreactor unit to dilute the crude protein and remove any air bubbles during mixing; and
(iii) a microfluidic purification unit, communicatively connected to the microfluidic mixer/de-bubbler unit, for capturing the crude protein and providing a purified protein, wherein the microfluidic purification unit comprises:
three polymethyl methacrylate (PMMA) layers comprising (a) a top layer, (b) a middle layer comprising at least one microcolumn, wherein each microcolumn has an inlet end and an outlet end, and (c) a base plate, wherein each microcolumn comprises chromatography resin, wherein the chromatography resin is retained in the microcolumn by a polytetrafluoroethylene (PTFE) frit positioned at the outlet end of the microcolumn, and

wherein all fluid channels of the device are microfluidic.

2. The factory-on-a-chip microfluidic device according to claim 1 , further comprising at least one process analytical technology (PAT) sensor for monitoring pH, pressure, temperature, dissolved-oxygen, redox conditions, ionic strength, UV-Vis absorbance, fluorescence, light scatter, and/or circular dichroism conditions during the reaction, purification and/or analysis of the crude and/or purified protein, wherein the at least one PAT sensor is communicatively connected to the microfluidic bioreactor unit and/or the at least one PAT sensor is communicatively connected to the microfluidic purification unit.

3. The factory-on-a-chip microfluidic device according to claim 1 , wherein the mixer/de-bubbler comprises a porous membrane to eliminate bubbles.

4. The factory-on-a-chip microfluidic device according to claim 1 , wherein the microfluidic bioreactor unit comprises cell extracts and a reaction mixture for cell-free expression of the crude protein, wherein a DNA or mRNA template is added to the cell extracts and the reaction mixture comprises biological extracts and/or reagents for expression of the crude protein.

5. The factory-on-a-chip microfluidic device according to claim 1 , wherein the microfluidic purification unit comprises 4 to 8 microcolumns.

6. The factory-on-a-chip microfluidic device according to claim 1 , wherein chromatography resin captures the crude protein.

7. The factory-on-a-chip microfluidic device according to claim 1 , wherein the chromatography resin is an immobilized metal affinity resin and an ion exchange resin.

8. The factory-on-a-chip microfluidic device according to claim 1 , wherein the microfluidic bioreactor unit comprises lyophilized cell extracts and reagents for expression of the crude protein.

9. The factory-on-a-chip microfluidic device according to claim 1 , wherein the microfluidic purification unit further comprises accommodations for an elution buffer for harvesting the purified protein.

10. The factory-on-a-chip microfluidic device according to claim 1 , wherein the microfluidic purification unit further comprises a PTFE frit positioned at the inlet end of the microcolumn.

11. The factory-on-a-chip microfluidic device according to claim 1 , wherein the at least one microcolumn of the purification column comprises microscale channels having a volume ranging from about 25-200 μL.

12. The factory-on-a-chip microfluidic device according to claim 1 , wherein the chromatography resin comprises an immobilized metal affinity resin.

13. The factory-on-a-chip microfluidic device according to claim 1 , wherein the chromatography resin comprises an ion exchange resin.

14. A method of preparing a purified therapeutic protein, said method comprising:
synthesizing and purifying the therapeutic protein in a factory-on-a-chip microfluidic device of claim 1 .

15. The method of claim 14 , further comprising administering the purified therapeutic protein to the subject, in a sufficient amount, within 24 hours of purification.

16. The method of claim 15 , wherein the purified therapeutic protein is refrigerated at a temperature above freezing from 0 to 6° C. before administration to the subject.

17. The method of claim 14 , wherein the microfluidic bioreactor unit comprises cell extracts and a reaction mixture for cell-free expression of the crude protein, wherein a DNA or mRNA template is added to the cell extracts and the reaction mixture comprises biological extracts and/or reagents for expression of the crude protein.

18. The method of claim 14 , wherein potency/activity of the purified therapeutic protein is at least 55% or more of the initial activity for at least 3 days at temperature from above 0° C. to about 30° C.

Assignees

• Univ Maryland

Inventors

• Rao Govind
• Kostov Yordan
• Andar Abhay
• Al-Adhami Mustafa
• Deldari Sevda
• Frey Douglas D

Classifications

• C12M41/40

  of pressure · CPC title

• C12M41/28

  of redox potential · CPC title

• C12M41/26

  of pH · CPC title

• C12M41/12

  of temperature (controlling the temperature of chemical or physical processes B01J19/0013, heating or cooling apparatus for laboratory use B01L7/00) · CPC title

• C12M33/14

  with filters, sieves or membranes · CPC title