Antigen-Presenting Synthetic Surfaces, Covalently Functionalized Surfaces, Activated T Cells, and Uses Thereof
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 activate T Lymphocytes.
1 . An antigen-presenting surface for activating a T lymphocyte (T cell), comprising:
a. a plurality of primary activating molecular ligands, wherein each primary activating molecular ligand comprises a major histocompatibility complex (MHC) Class I molecule configured to bind to a T cell receptor (TCR) of the T cell; and,
b. a plurality of co-activating molecular ligands each comprising a TCR co-activating molecule or an adjunct TCR activating molecule,
wherein each of the plurality of primary activating molecular ligands and the plurality of co-activating molecular ligands is specifically bound to the antigen presenting surface, and
wherein the antigen presenting surface is the surface of a bead.
2 . The antigen-presenting surface of claim 1 , wherein the ratio of the TCR co-activating molecules to the adjunct TCR activating molecule is about 20:1 to about 1:20.
3 . The antigen-presenting surface of claim 1 , wherein the plurality of primary activating molecular ligands is disposed upon at least a portion of a surface of the antigen-presenting surface at a density from about 4×10 2 to about 3×10 4 molecules per square micron.
4 . The antigen-presenting surface of claim 1 , further comprising a plurality of surface-blocking molecular ligands.
5 . The antigen-presenting surface of claim 1 , wherein the plurality of co-activating molecular ligands is disposed upon at least a portion of a surface of the antigen-presenting surface at a density from about 5×10 3 to about 2×10 4 molecules per square micron or about 5×10 3 to about 1.5×10 4 molecules per square micron.
6 . The antigen-presenting surface of claim 1 , wherein a ratio of the primary activating molecular ligands to the co-activating molecular ligands present on the antigen-presenting surface is about 1:10 to about 2:1.
7 . The antigen-presenting surface of claim 1 , wherein:
(i) the antigen-presenting surface comprises glass, metal, ceramic, and/or a metal oxide; or
(ii) the antigen-presenting surface comprises a polymeric surface.
8 . The antigen-presenting surface of claim 1 , wherein each of the plurality of primary activating molecular ligands is noncovalently bound to a binding moiety, and further wherein the binding moiety is (i) covalently bound to the antigen-presenting surface or (ii) noncovalently bound to a second binding moiety that is covalently bound to the surface.
9 . The antigen-presenting surface of claim 1 , wherein the MHC molecule comprises an MHC protein sequence and a beta microglobulin.
10 . The antigen-presenting surface of claim 1 , wherein the MHC molecule further comprises a tumor associated antigen.
11 . The antigen-presenting surface of claim 10 , wherein the tumor associated antigen is SLC45A2, TCL1, VCX3A, MART1 or NYESO1.
12 . The antigen-presenting surface of claim 1 , wherein:
(i) the TCR co-activating protein molecule comprises a CD-28 binding protein or a fragment thereof which retains binding ability with CD28; or
(ii) the first TCR co-activating molecule comprises an anti-CD28 antibody or a fragment thereof, wherein the fragment retains binding activity with CD28.
13 . The antigen-presenting surface of claim 1 , wherein:
(i) the adjunct stimulatory molecule comprises a CD2 binding protein or a fragment thereof, wherein the fragment retains binding activity with CD2; or
(ii) the adjunct stimulatory molecule comprises an anti-CD2 antibody or a fragment thereof, wherein the fragment retains binding activity with CD2.
14 . The antigen-presenting surface of claim 4 , wherein each of the plurality of surface-blocking molecular ligands comprises a polyethylene (PEG) moiety, a carboxylic acid moiety, or a combination thereof.
15 . The antigen-presenting surface of claim 1 , wherein the ratio of the TCR co-activating molecules to the adjunct TCR activating molecules of the plurality of co-activating molecular ligands is from about 3:1 to about 1:3.
16 .- 60 . (canceled)
61 . The antigen-presenting surface of claim 1 , wherein the surface comprises a modified surface functionalized with a functionality comprising an azide, a carboxylic acid or active ester thereof, a succinimide ester, a maleimide, a keto, a sulfonyl halide, sulfonic acid, dibenzocyclooctyne, an alkene, or an alkyne.
62 . The antigen-presenting surface of claim 1 , wherein the surface comprises a modified surface functionalized with azide reactive moieties.
63 . The antigen-presenting surface of claim 62 , wherein the azide reactive moieties are selected from an alkyne, tetrabutylammonium azide, and Formula 1:
64 . The antigen-presenting surface of claim 62 , wherein the modified functionalized surface is reacted with a streptavidin moiety or a biotin moiety.
65 . The antigen-presenting surface of claim 64 , wherein the modified functionalized surface is reacted with a polyethylene glycol (PEG) moiety, a carboxylic acid moiety, or a combination thereof.