Granted Patent B2

US 12,251,674 · App. 16/928,723 · Granted Mar 18, 2025

Inventors: John J. Rajasekaran (Hillsborough, CA); Vasanth Jayaraman (San Mateo, CA); Tianhao Wang (San Mateo, CA); Kang Bei (San Mateo, CA); Hari Krishnan Krishnamurthy (San Mateo, CA); Punitha Vedantham (San Carlos, CA)

Assignee: Vibrant Holdings, LLC

B01J19/0046C40B50/18G01N33/6842B01J2219/00596B01J2219/00637B01J2219/00711B01J2219/00725

Abstract

Claims (18)

1. A method of attaching a coupling molecule to a substrate, comprising:

obtaining a substrate comprising a plurality of carboxylic acid groups for linking to a coupling molecule;

contacting said substrate with a carboxylic acid activating compound formulation comprising a carbodiimide precursor selected from the group consisting of: 1-(3-(diethylamino)-propyl)-4-(2-methoxyphenyl)-1,4-dihydro-5H-tetrazole-5-thione, 4,5-dihydro-4-(hydroxymethyl)-1-phenyl-1H-tetrazole-5-thione, 1-(3-(dimethylamino) propyl)-4-ethyl-1,4-dihydro-5H-tetrazole-5-thione, 1,4-Bis (2,2-dimethyl-1,3-dioxolan-4-ylmethyl)-1,4-dihydro-5H-tetrazole-5-thione, 4-cyclohexyl-1H-tetrazole-5(4H)-thione, and 1-phenyl-4-(piperidinomethyl)-tetrazole-5(4H)-thione, wherein said carbodiimide precursor converts to a carbodiimide upon exposure to electromagnetic radiation at a defined wavelength;

selectively exposing said carboxylic acid activating compound formulation to the electromagnetic radiation, thereby generating said carbodiimide and activating the carboxylic acid groups to form carbonyl groups on said substrate at a selectively exposed area;

contacting said substrate with a coupling formulation comprising a coupling molecule and a solvent;

coupling the coupling molecule to at least one of said plurality of carbonyl groups at said selectively exposed area.

2. The method of claim 1 , wherein said defined wavelength is 248 nm or 193 nm.

3. The method of claim 1 , wherein said carboxylic acid activating compound formulation or said coupling formulation further comprises a polymer.

4. The method of claim 3 , wherein said polymer is poly (methyl methacrylate), polyvinyl pyrrolidone, or polyvinyl alcohol.

5. The method of claim 1 , wherein said carboxylic acid activating compound formulation and said coupling formulation are present on the substrate when said substrate is exposed to said electromagnetic radiation.

6. The method of claim 1 , wherein said coupling step is performed multiple times at different selectively exposed areas on said substrate.

7. The method of claim 1 , wherein said coupling step has a coupling efficiency of at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 98.5%, or 99%.

8. The method of claim 1 , wherein said coupling molecule is an amino acid, a protein, or a polypeptide.

9. The method of claim 1 , wherein said coupling molecule comprises a protected amine group.

10. The method of claim 9 , wherein said amine group is protected by Fmoc.

11. The method of claim 1 , further comprising repeating said method to produce a desired polypeptide at said at least one carbonyl group.

12. The method of claim 1 , wherein said substrate comprises a porous layer, and optionally wherein said substrate comprises a porous layer comprising a plurality of attachment sites extending in multiple dimensions from the surface of said porous layer within and around said porous layer.

13. The method of claim 1 , wherein said substrate comprises a planar layer comprising a metal and having an upper surface and a lower surface; and a plurality of pillars operatively coupled to the layer in positionally-defined locations, wherein each pillar has a planar surface extended from the layer, wherein the distance between the surface of each pillar and the upper surface of the layer is between 1,000-5,000 angstroms, wherein the surface of each pillar is parallel to the upper surface of the layer, and wherein the plurality of pillars are present at a density of greater than 10,000/cm 2 , and wherein the attachment site is coupled to the upper surface of the pillar.

Assignments (2)

Continuity (11)

References Cited (335)