Covalently modified surfaces, kits, and methods of preparation and use
Inventors
Lowe, JR., Randall D. • MASTROIANNI, Alexander J. • White, Mark P. • Lavieu, Gregory G. • Beaumont, Kristin G.
Assignees
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Publication Number
US-12280370-B2
Publication Date
2025-04-22
Expiration Date
Abstract
In biosciences and related fields, it can be useful to modify surfaces of apparatuses, devices, and materials that contact biomaterials such as biomolecules and biological micro-objects. Described herein are surface modifying and surface functionalizing reagents, preparation thereof, and methods for modifying surfaces to provide improved or altered performance with biomaterials.
Core Innovation
The invention provides a kit that includes a microfluidic device with an enclosure comprising a base, a cover, and microfluidic circuit material defining a fluidic circuit. At least one inner surface of the base, the cover, and the microfluidic circuit material has a plurality of first covalently bound surface modifications and a plurality of second covalently bound surface modifications, where each modification includes a linking group and a moiety that is a reactive moiety or a surface contact moiety.
The kit also includes a first surface modifying reagent having a structure of Formula XII or Formula XII1. The reagent includes a reaction pair moiety RP1, a surface contact moiety configured to support cell growth, viability, portability, or any combination thereof, and a linker L1 that is a bond or a chain of 1 to 200 non-hydrogen atoms selected from silicon, carbon, nitrogen, oxygen, sulfur and phosphorus atoms, and may further include 0 or 1 coupling groups CG.
The disclosed surface-functionalizing chemistries use covalently modified surfaces and orthogonal reaction pair coupling for biological functionalization and cell-compatible surface modification. Embodiments further include substantially all fluid-facing interior microfluidic surfaces, methods of preparing covalently modified microfluidic device surfaces with first and second distinct covalent modifications, and kit components that include the microfluidic device plus surface modifying reagents.
Claims Coverage
The consolidated claim coverage centers on one independent claim for a kit combining a microfluidic device with first and second covalently bound surface modifications and a first surface modifying reagent defined by Formula XII or Formula XII1. The inventive features are the microfluidic device architecture and the defined reagent with RP1, a cell-supporting surface contact moiety, and a specified linker L1.
Microfluidic device with covalently bound first and second surface modifications
A microfluidic device comprising an enclosure with a base, a cover, and microfluidic circuit material defining a fluidic circuit, where inner surfaces have a plurality of first covalently bound surface modifications with a first linking group and a first moiety, and a plurality of second covalently bound surface modifications with a second linking group and a second moiety.
First surface modifying reagent defined by Formula XII or Formula XII1
A first surface modifying reagent having a structure of Formula XII or Formula XII1 including RP1 as a reaction pair moiety, a surface contact moiety configured to support cell growth, viability, portability, or any combination thereof, and a linker L1 that is a bond or a chain of 1 to 200 non-hydrogen atoms selected from silicon, carbon, nitrogen, oxygen, sulfur, and phosphorus atoms, optionally including 0 or 1 coupling groups CG.
Distinct second Formula XII reagent and constrained reactive moieties
A second surface modifying reagent defined by Formula XII with a second reaction pair moiety RP2, a second surface contact moiety, and a second linker L2, where the second surface modifying reagent is distinct from the first surface modifying reagent. Dependent claims also constrain reactive moieties to selected functional-group classes and selected formula structures.
The claim scope is defined by a kit architecture that combines a microfluidic device with internal first and second covalently bound surface modifications and a Formula XII reagent system. The inventive features combine defined linking groups and moieties with a surface contact moiety configured to support cell growth, viability, or portability, and further refine the reagent and reactive moiety structures.
Stated Advantages
Supports cell growth, viability, portability, or any combination thereof through the surface contact moiety.
Provides covalently bound surface modifications on inner surfaces of the microfluidic device, including the base, cover, and microfluidic circuit material.
Enables multiple surface modification moieties using first and second covalently bound surface modifications.
Enables ionic interactions between modified-surface ions and charged noncovalent proteins in a fluid medium.
Supports cell viability and functional cell growth.
Documented Applications
Support cell growth, viability, portability through microfluidic device surface modifications.
Cell growth/viability in a microfluidic device having covalently modified internal surfaces.
Use of covalently modified microfluidic internal surfaces to enable ionic interactions with charged noncovalent proteins in a fluid medium.
Microfluidic sequestration pens.
Cell adherence and cell-culture utility, including HeLa cell adherence, growth, multiplication, and support on PEG/PLL-type positively charged surfaces and mixed PEG/pol-L-lysine layers.
Regioselective surface modification in a microfluidic channel, including confinement of PEG5K to a microfluidic channel and localization of poly-L-lysine to sequestration pens.
Biological functionalization using streptavidin/biotin to attach fibronectin, and optionally laminin.
Surface functionalization with mixed biotin/PEG surfaces and Streptavidin binding, including thickness measurements versus biotin/SAV ratio.
Generation and evaluation of covalently modified surfaces with physical characterization such as layer thickness and water contact angles, including monolayer uniformity.
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