Functionalized well plate, methods of preparation and use thereof
Inventors
Beemiller, Peter J. • MASTROIANNI, Alexander J. • Lowe, JR., Randall D. • Bronevetsky, Yelena
Assignees
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Publication Number
US-11993766-B2
Publication Date
2024-05-28
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 of wells within a well plate to activate lymphocytes, including but not limited to T lymphocytes, in a controllable and reproducible manner.
Core Innovation
The invention relates to a well plate comprising a surface for presenting an antigen. The surface includes a first region that is an activating moiety-presenting covalently functionalized region with a plurality of specifically bound primary activating molecular ligands and a plurality of specifically bound co-activating molecular ligands. The first region has an area of 2 mm^2 to 35 mm^2.
The described surface architecture includes covalently functionalized regions that present immunologically relevant ligands in a spatially defined region size. Each ligand contains a major histocompatibility complex (MHC) molecule configured to bind a T cell receptor (TCR) on a T cell, and the co-activating molecular ligands provide co-activation for the TCR pathway.
The document also describes a kit for preparing the well plate, where the surface includes a reactive moiety-presenting covalently functionalized region having an area of 2 mm^2 to 35 mm^2. The reactive moiety is selected from an azido moiety, an alkynyl moiety, a biotin moiety, or a streptavidin moiety, and the kit further includes an activating reagent comprising a primary activating reagent and a co-activating reagent.
Further disclosed aspects include use of a second surface region or a surface blocking region containing surface blocking ligands to surround and reduce unwanted attachment and biofouling.
Claims Coverage
The document provides two independent claims covering a functionalized antigen-presenting well plate and a kit for preparing such a well plate. Across these claims, there are two principal groups of inventive features: covalently functionalized activating regions presenting both primary activating ligands and co-activating ligands, and kit elements that pair a reactive moiety-presenting covalently functionalized region with an activating reagent containing primary and co-activating components, each constrained to an activating region area of 2 mm^2 to 35 mm^2.
Activating moiety-presenting covalently functionalized first region presenting primary and co-activating molecular ligands
A well plate comprising a surface for presenting an antigen, wherein the surface has a first region which is an activating moiety-presenting covalently functionalized region comprising a plurality of specifically bound primary activating molecular ligands and a plurality of specifically bound co-activating molecular ligands.
Defined activating region area
The first region has an area of 2 mm^2 to 35 mm^2.
Reactive moiety-presenting covalently functionalized region as a kit component
A kit for preparing a well plate comprising a surface for presenting an antigen, wherein the surface has an activating moiety-presenting covalently functionalized region, comprising a well plate comprising a surface comprising a reactive moiety-presenting covalently functionalized region, wherein the reactive moiety is an azido moiety, an alkynyl moiety, a biotin moiety, or a streptavidin moiety.
Activating reagent with primary activating reagent and co-activating reagent
A kit for preparing a well plate further comprising an activating reagent comprising a primary activating reagent and a co-activating reagent.
Defined reactive/activating region area in the kit
The reactive moiety-presenting covalently functionalized region has an area of 2 mm^2 to 35 mm^2.
Overall, the independent claim coverage centers on covalently functionalized, area-defined activating regions on a well plate that present both primary activating and co-activating molecular ligands, and on a kit that prepares such a plate by combining a reactive moiety-presenting covalently functionalized region with an activating reagent having primary and co-activating reagents.
Stated Advantages
Reduce unwanted attachment and biofouling.
Documented Applications
A well plate comprising a surface for presenting an antigen.
A kit for preparing such a well plate.
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