Aqueous biomolecule coupling on CO2-plasma-activated surfaces

Patent metadata

Field	Value
Publication number	US-11919971-B2
Application number	US-201716302023-A
Country	US
Kind code	B2
Filing date	May 16, 2017
Priority date	May 16, 2016
Publication date	Mar 5, 2024
Grant date	Mar 5, 2024

Abstract

This disclosure provides, e.g., methods for coupling Formation of Surface Carboxylates on PES an entity to a solid substrate. The method can comprise treating the with Exposure Time substrate with a plasma, e.g., a CO2 plasma, to increase its reactivity. The entity can be, e.g., a biological polymer that binds a microbe. Substrates produced by these methods can be used in a variety of applications, including hemodialysis and diagnostic assays.

First claim

What is claimed is:

1. A method of making a substrate having an entity attached thereto, the method comprising:
i) contacting the substrate with a CO 2 plasma to form a modified substrate comprising a plasma-generated-moiety (PGM) comprising a carboxylic acid group, wherein the substrate comprises polysulfone (PS), polyarylethersulfone (PAES) or polyethersulfone (PES); and ii) contacting the entity with the modified substrate under conditions sufficient for attachment of the entity to the modified substrate; wherein step ii) contacting is performed for about 4-16 hours;

thereby making a substrate having the entity attached thereto,
provided that
the entity comprises a lectin; and

provided that:
the modified substrate is not contacted with a crosslinking moiety of derivatized with a crosslinking moiety, prior to attachment of the entity;
the modified substrate is not contacted with an organic solvent prior to attachment of the entity; and
the entity is contacted with the modified substrate under aqueous conditions.

2. The method of claim 1 , wherein the substrate comprises a lumen.

3. The method of claim 1 , wherein the substrate comprises an adhesive or a sealant, and wherein the adhesive or sealant is not contacted with an organic solvent.

4. The method of claim 1 , wherein the substrate comprises a dialysis, ultrafiltration, hemofiltration, hemodiafiltration, or hemoperfusion cartridge.

5. The method of claim 1 , wherein in step (ii), the modified substrate is substantially free of a crosslinking moiety.

6. The method of claim 1 , wherein in step (ii), the modified substrate is substantially free of organic solvent, or wherein the method does not comprise a step of contacting the modified substrate with an organic solvent.

7. The method of claim 1 , wherein the method comprises
contacting the modified substrate, the entity, or both, with an activating moiety to activate a functional group on the modified substrate, wherein the functional group is a carboxylic acid group.

8. The method of claim 1 , wherein
step ii) contacting is performed at a pH of 4.

9. The method of claim 1 , wherein the PGM comprises a carboxylic acid and the entity comprises an amine and wherein a carboxylic acid of the PGM covalently binds with an amine group of the entity.

10. The method of claim 1 , wherein at least 1% of the PGMs comprise a carboxylic acid group.

11. The method of claim 1 , wherein step 1, contacting comprises contacting a plurality of substrates with the plasma in a vacuum chamber of a radio frequency plasma generator.

12. The method of claim 1 , wherein the entity comprises a polypeptide of SEQ ID NO: 4 or at least 80% identical to SEQ ID NO: 4, or a polypeptide of SEQ ID NO: 6 or at least 80% identical to SEQ ID NO: 6.

13. A device comprising a substrate having an entity attached thereto, produced by the method of claim 1 .

14. The method of claim 1 , wherein the plasma consists essentially of CO 2 plasma or consists of CO 2 plasma.

15. The method of claim 3 wherein the PGM comprises an aldehyde group.

16. The method of claim 1 , wherein the plasma treatment step is less than 10 minutes.

17. The method of claim 1 , wherein the plasma is generated by a plasma generator under a radio frequency of about 12 MHz to 15 MHz.

18. The method of claim 17 , wherein the plasma generator comprises electrodes outside a vacuum chamber of the plasma generator.

19. The method of claim 1 , wherein step i) contacting comprises contacting the substrate with the plasma in a vacuum chamber of a radio frequency plasma generator, and the plasma is generated by the plasma generator under a radio frequency of about 12 MHz to 15 MHz.

20. The method of claim 1 , wherein the entities are attached at a density of at least about 1×10 12 molecules per cm 2 .

21. The method of claim 1 , wherein the contacting of step ii) is performed in a solution comprising 2-morpholino-ethane sulfonic acid (MES) buffer.

22. The method of claim 1 , wherein step ii) contacting is performed for about 4-6 hours.

Assignees

• Harvard College

Inventors

• Leslie Daniel Christopher
• Doyle Thomas
• Waterhouse Anna
• Rodas Melissa
• Watters Alexander L
• Super Michael
• Ingber Donald E

Classifications

• C07K17/08 Primary

  the carrier being a synthetic polymer · CPC title

• C08L25/06

  Polystyrene · CPC title

• C08L83/04

  Polysiloxanes · CPC title

• G01N33/54393

  Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding · CPC title

• G01N33/545

  Synthetic resin · CPC title