WO2023114125A1

WO2023114125A1 - Detection device and methods of use thereof

Info

Publication number: WO2023114125A1
Application number: PCT/US2022/052515
Authority: WO
Inventors: Abigail BARNES; Lukasz MENDECKI
Original assignee: Amulet, Inc.
Priority date: 2021-12-13
Filing date: 2022-12-12
Publication date: 2023-06-22

Abstract

A detection device includes a sensor comprising a circuit board, an electropolymerized molecularly imprinted polymer ("MIP") film that includes receptor sites imprinted in the polymer, the receptor sites configured to accept a molecule, part of a molecule, and/or a combination of molecules, and an electropolymerized non-imprinted polymer ("NIP") film. The sensor is configured to detect the presence of the molecule in a sample, when the sensor is exposed to that sample, and when the molecule binds to one or more of the receptor sites on the MIP and the electropolymerized MIP and NIP films include one or more polymerized monomers selected from formulas (I), (II), (III), (IV), (V), (VI), (VII), (VIII), and (IX) or 3, 4-ethylenedi oxythiophene.

Description

DETECTION DEVICE AND METHODS OF USE THEREOF

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63/288,867, filed December 13, 2021, the contents of which are incorporated herein by reference in their entirety.

FIELD

[0002] The present technology is directed to detection devices including electropolymerized molecularly imprinted polymer (“MIP”) films and electropolymerized non-imprinted polymer (“NIP”) films. In particular, the detection devices may be used to detect various molecules, including, e.g., an allergen, a trace molecule of an allergen, a pathogen, a toxin, or a combination of any two or more thereof.

SUMMARY

[0003] In one aspect, a detection device is provided. The device may include a sensor that includes an MIP film comprising receptor sites imprinted in the polymer, wherein the receptor sites are configured to accept a molecule, part of a molecule, and/or a combination of molecules; and an NIP film (z.e., the control). In any embodiment, the sensor may be configured to detect the presence of the molecule upon binding to the one or more receptor sites. In any embodiment, the MIP film and the NIP film may include one or more polymerized monomers selected from the group consisting of 3, 4-ethylenedi oxythiophene or a monomer of formula I, II, III, IV, V, VI, VII, VIII, or IX:

formula IX wherein: A, B, D, E, and G are independently CH or N, provided that at least one of A, B, D, E, and G are N; J is N or S; R¹, R², and R³ are independently H or NH2; R⁴, R⁵, and R⁷ are independently

absent, H, or CH3; R¹⁰ and R¹¹ are independently H or C1-C5 alkylene-NH2; R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ are independently H, NH2, and OH, provided that one of R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ is NH2 or OH; R¹⁵ C1-C5 alkyl; R¹⁶, R¹⁷, and R¹⁸ are independently H, NH2, or SH; and m is 1, 2, 3, 4 or 5. In any embodiment, the molecule may include an allergen, a trace molecule of an allergen, a pathogen, a toxin, or a combination of any two or more thereof.

[0004] In any embodiment, the device may further include one or more electrochemical chips. In some embodiments, the device may include one electrochemical chip comprising the MIP film and the NIP film. In some embodiments, the device may include a first electrochemical chip, wherein the first electrochemical chip comprises the MIP film and/or a second electrochemical chip, wherein the second electrochemical chip comprises the NIP film. In any embodiment, the device may further include a circuit board (e.g., printed circuit board) comprising the first electrochemical chip and the second electrochemical chip.

[0005] In any embodiment, the device may further include a processing device. Commonly, the processing device may be configured to communicatively couple to the sensor and may be configured to determine an electric current and/or potential difference between the electropolymerized MIP film and the electropolymerized NIP film. In any of the above embodiments, the processing device may determine the presence of the molecule when the electric current and/or potential difference of the electropolymerized MIP film is greater than the electric current and/or potential difference of the electropolymerized NIP film. In any of the above embodiments, the processing device may determine the presence of the molecule when the electric current and/or potential difference of the electropolymerized MIP film is lower than the electric current and/or potential difference of the electropolymerized NIP film.

[0006] In any of the above embodiments, the device may also include a body, wherein the body comprises a capsule that encapsulates a solvent; and a chamber for mixing the solvent with a tangible good sample that may include the molecule. In any embodiment, the device may further include a barrier, wherein puncturing the barrier exposes the mixture of solvent and tangible good sample to the electropolymerized MIP film and electropolymerized NIP film.

[0007] In another aspect, a detection device comprises a sensor, wherein the sensor comprises: a circuit board; a molecularly imprinted polymer (MIP) film comprising one or more receptor sites imprinted in the polymer, the one or more receptor sites configured to accept a target molecule; and a non-imprinted polymer (NIP) film; wherein the sensor is configured to detect the presence of the target molecule in a sample, when the sensor is exposed to the sample, upon binding to one or more of the receptor sites; and the MIP film and the NIP film comprise one or more polymerized monomers selected from the group consisting of 3, 4-ethylenedi oxythiophene or a monomer of formula I, II, III, IV, V, VI, VII, VIII, or IX, as defined herein. In any of the embodiments herein, the MIP film may have a thickness of about 1 nm to about 100 nm, such as about 2 nm to about 20 nm. In any of the above embodiments or aspects, the target molecule may be an epitope.

[0008] In a further aspect, a process for preparing a detection device includes depositing on a sensor a template molecule on a portion of a surface of the sensor such that the template molecule forms a self-assembled monolayer on the sensor in a substantially perpendicular orientation to the surface to form a modified sensor; polymerizing a polymer film on the modified sensor forming both imprinted regions where the template molecule is present and non-imprinted regions where the template molecule is absent; and removing the template molecule from the sensor, such that a cavity is formed where the template molecule was present, the cavity being a specific binding pocket for a target molecule of the same as or similar to the template molecule; wherein the polymer film is the electropolymerization product of one or more of 3, 4-ethylenedi oxythiophene, or a monomer of formula I, II, III, IV, V, VI, VII, VIII, or IX:

formula IX

In the formulae, A, B, D, E, and G are independently CH or N, provided that at least one of A, B, D, E, and G are N; J is N or S; R¹, R², and R³ are independently H or NH2; R⁴, R⁵, and R⁷ are independently

is absent, H, or CH3; R¹⁰ and R¹¹ are independently H or C1-C5 alkylene-NH2; R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ are independently H, NH2, and OH, provided that one of R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ is NH2 or OH; R¹⁵ C1-C5 alkyl; R¹⁶, R¹⁷, and R¹⁸ are independently H, NH2, or SH; and m is 1, 2, 3, 4 or 5. In any embodiments herein, the polymer film may have a thickness of about 1 nm to about 100 nm, or about 2 nm to about 20 nm. In some embodiments of the methods, it may include swelling the film to allow release of the template molecule. In any of the above embodiments, the removing of the method includes cleaving a bond between a terminal end of the template molecule and the surface, such that the cavity is of sufficient dimensions and shape to be specific to the target molecule. In any of the above embodiments, the template molecule may include an epitope. In such cases, the cleaving of a bond includes cleaving a molecule to surface bond, or a disulfide bond associated with the epitope to remove a portion of the template molecule.

DETAILED DESCRIPTION

[0009] The present technology provides a fast and portable detection device enabling users to directly sample tangible goods (e.g., foods) for unwanted molecule(s) (e.g., allergens, pathogens, and/or toxins). For example, the device may provide individuals with information about the contents and relative safety of their food and other tangible goods. The presence or absence of the molecule may be detected by capturing a representative sample, such as a liquid or solid goods sample and exposing the sample to the sensor. The sensor may then be connected to a processing device (e.g., Amulet). The sensor includes the MIP with electropolymerized detection (together referred to as an electropolymerized MIP). MIPs are polymer compositions having synthetic cavities, or binding pockets, designed to bind to the molecules. If the molecule is present in the tested sample, binding occurs, i.e. the target molecule or a molecule indicative of the target molecule fills the binding pocket in the MIP, and the processing device then detects a measurable interaction, alerting the user to the presence of the molecule within a short period of time (e.g., seconds). If no binding occurs, the processing device signals that the molecule was not detected.

[0010] The processing device can be configured as a wearable device, or integrated into everyday products that users may keep on their person (e.g., cellular phone, watch, keychain, necklace, etc.). A software application (i.e. “app”), may accompany the processing device, where users may track and upload tests, connect with other users, and store and share important information including, but not limited to, emergency contacts.

[0011] The following terms are used throughout this disclosure, as defined below. [0012] As used herein and in the appended claims, singular articles such as “a” and “an” and “the” and similar referents in the context of describing the elements (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the embodiments and does not pose a limitation on the scope of the claims unless otherwise stated. No language in the specification should be construed as indicating any non-claimed element as essential.

[0013] As used herein, “about” will be understood by persons of ordinary skill in the art and will vary to some extent depending upon the context in which it is used. If there are uses of the term which are not clear to persons of ordinary skill in the art, given the context in which it is used, “about” will mean up to plus or minus 10% of the particular term.

[0014] Unless otherwise indicated, numeric ranges, for instance as in “from 2 to 10,” are inclusive of the numbers defining the range (e.g., 2 and 10).

[0015] Unless otherwise indicated, ratios, percentages, parts, and the like are by weight.

[0016] Alkyl groups include straight chain and branched chain alkyl groups having from 1 to 12 carbon atoms, and typically from 1 to 10 carbons or, in some embodiments, from 1 to 8, 1 to 6, or 1 to 4 carbon atoms. Examples of straight chain alkyl groups include groups such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, secbutyl, tert-butyl, neopentyl, isopentyl, and 2,2-dimethylpropyl groups. Representative substituted alkyl groups may be substituted one or more times with substituents such as those listed above, and include without limitation haloalkyl (e.g., trifluoromethyl), hydroxyalkyl, thioalkyl, aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl, alkoxyalkyl, carboxyalkyl, and the like.

[0017] Aryl groups are cyclic aromatic hydrocarbons that do not contain heteroatoms. Aryl groups herein include monocyclic, bicyclic and tricyclic ring systems. Thus, aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, fluorenyl, phenanthrenyl, anthracenyl, indenyl, indanyl, pentalenyl, and naphthyl groups. In some embodiments, aryl groups contain 6-14 carbons, and in others from 6 to 12 or even 6-10 carbon atoms in the ring portions of the groups. In some embodiments, the aryl groups are phenyl or naphthyl. Although the phrase “aryl groups” includes groups containing fused rings, such as fused aromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, and the like), it does not include aryl groups that have other groups, such as alkyl or halo groups, bonded to one of the ring members. Rather, groups such as tolyl are referred to as substituted aryl groups. Representative substituted aryl groups may be mono-substituted or substituted more than once, e.g., 2, 3, 4, or 5 times. Monosubstituted aryl groups include, but are not limited to, 2-, 3-, 4-, 5-, or 6-substituted phenyl or naphthyl groups, which may be substituted with substituents such as those listed above.

[0018] Groups described herein having two or more points of attachment (i.e., divalent, trivalent, or polyvalent) within the compound of the present technology are designated by use of the suffix, “ene.” For example, divalent alkyl groups are alkylene groups, divalent aryl groups are arylene groups, divalent heteroaryl groups are heteroarylene groups, and so forth. Substituted groups having a single point of attachment to the compound of the present technology are not referred to using the “ene” designation. Thus, e.g., chloroethyl is not referred to herein as chloroethylene.

[0019] The term “hydroxyl” as used herein can refer to -OH or its ionized form, -O“. A “hydroxyalkyl” group is a hydroxyl-substituted alkyl group, such as HO-CH2-.

[0020] The term “amine” (or “amino”) as used herein refers to -NR⁷⁵R⁷⁶ groups, wherein R⁷⁵ and R⁷⁶ are independently hydrogen, or a substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl or heterocyclyl group as defined herein. In some embodiments, the amine is alkylamino, dialkylamino, arylamino, or alkylarylamino. In other embodiments, the amine is NH2, methylamino, dimethylamino, ethylamino, diethylamino, propylamino, isopropylamino, phenylamino, or benzylamino.

[0021] The term “aromatic” as used herein refers to a group in an organic molecule that is cyclic, planar, and has pi bonds in resonance that gives increased stability compared to other geometric or connective arrangements with the same set of atoms.

[0022] The term “complexation energy” refers to the relative strength of the intermolecular interactions between the polymerized monomer and the molecule. For a polymerized monomer with energy E_m, target with energy E_t, and polymerized monomer- molecule complex energy E_mt, all calculated at the same level of theory, the complexation energy E_c is given by:

E_c ⁼ Emt ~ E_m + E_t)

[0023] As used herein “electric potential difference” or “potential difference” or “voltage” refers to the difference in electric potential between two points. If there is no potential difference then there won’t be flow of charge or “electric current” or “current”. Electric potential different and electric current are directly proportional and related by the following equation:

I=AV/R where I is current, AV is electric potential difference, and R is resistance. In any embodiment, electric current and/or potential difference may be directly measured, determined through a mathematical construct (e.g., average or weighted average), or a combination thereof. Commonly, electric current and/or potential difference measurements may be taken by any known electrochemical experiment including, but not limited to, cyclic voltammetry (CV), linear sweep voltammetry, square wave voltammetry, differential pulse voltammetry, amperometry, or a combination thereof. In any embodiment, electric current and/or potential difference may be measured at the maximum, minimum, or average current and/or potential difference between pre-set voltage values and/or between inflection points. These measurements may be taken before and after incubation, continuously, or with some degree of mid-incubation data points. In any embodiment, changes between the preincubation and post-incubation measurements for the MIP and NIP control films indicate the presence or absence of the target molecule. Pre-incubation measurements may be stored in memory as a set value or as bar codes, QR codes, or in flash drives. If measurements are taken in a combined sample/electrolyte incubation solution, continuous/intermediate scans may be taken as well. Many uses can be envisioned for intermediate data points, including checking the veracity of the pre-incubation and post-incubation measurements to enhance test confidence. Other data, such as points of maximum current, average current, an inflection point, and/or at a pre-defined voltage during either the oxidative or reductive phase of a CV may be used to compare pre- and post-incubation results. This data may be used in place of, or to supplement, peak current measurements. In any embodiment, electric currents and/or potential differences may be derived from a single cycle of an electrochemical experiment or an average or weighted average from two or more cycles.

[0024] The present technology provides a detection device including a sensor, wherein the sensor comprises a circuit board; an electropolymerized MIP film comprising one or more receptor sites imprinted in the polymer, the one or more receptor sites configured to accept a molecule, part of a molecule, and/or a combination of molecules; and an electropolymerized NIP film. The MIP and NIP may be in different regions of the same sensor, with the NIP acting as a control during operation of the detection device. The electropolymerized MIP film and the electropolymerized NIP film includes one or more polymerized monomers selected from the group consisting of 3, 4-ethylenedi oxythiophene or a monomer of formula I, II, III, IV, V, VI, VII, VIII, or IX:

wherein: A, B, D, E, and G are independently CH or N, provided that at least one of A, B, D, E, and G are N; J is N or S; R¹, R², and R³ are independently H or NH2; R⁴, R⁵, and R⁷ are independently

absent, H, or CH3; R¹⁰ and R¹¹ are independently H or C1-C5 alkylene-NH2; R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ are independently H, NH2, and OH, provided that one of R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ is NH2 or OH; R¹⁵ C1-C5 alkyl; R¹⁶, R¹⁷, and R¹⁸ are independently H, NH2, or SH; and m is 1, 2, 3, 4 or 5.

[0025] In any embodiment, the molecule may be in a tangible good. As used herein “tangible goods” or “good” or “goods” or “good sample” refers to physical goods (i.e., goods that can be touched). In any embodiment, the molecule may be in a manufactured good or consumable good. As used herein “manufactured goods” refers to goods that are a result of manufacturing (i.e., taking starting materials and transforming them into a product). Manufactured goods may be produced on a large or small scale. In any embodiment, the molecule may be in a consumable good. As used herein “consumable goods” refers to goods that are intended to be consumed. Manufactured goods may include consumable goods. In any embodiment, the consumable good may include food, drink, personal care (e.g., cosmetic such as skincare or haircare product such as cleanser, moisturizer, shampoo, conditioner, makeup, and/or perfume), or a combination of two or more thereof. A tangible good may include one or more of the molecules. A “molecule” also referred to herein as a “template molecule” or “target molecule” refers to a molecule that can be used to create receptor sites in the polymer to create the electropolymerized MIP film. The molecule may be present in any of a variety of items that may be a target for detecting an allergen, pathogen, and/or toxin. For example, a molecule may be present in the tangible good itself or in an item the tangible good has come into contact. For example, an item that a food has come into contact with (e.g., a serving utensil, a table, etc.) or an item that a skincare product has come into contact (e.g., plastic packaging). Tangible goods that may include a molecule may come in a variety of forms including, but not limited to, a solid, a liquid, a gas, a suspension, an emulsification, and any combinations thereof. Example solid tangible goods include, but are not limited to, a solid food (e.g., a bread, a nut), a plate, a table, a utensil, solid makeup (e.g., eyeshadow or lipstick), and any combinations thereof. Example liquid tangible goods include, but are not limited to, a liquid food, a beverage (e.g., a soda, milk, a juice), a food extract, shampoo, perfume, and any combinations thereof. Examples of suspension tangible goods include, but are not limited to, a tangible good suspended in air (e.g., a composition in particulate form), a tangible good suspended in a solvent (e.g., sprayable hair product), and any combinations thereof. Examples of emulsion tangible goods include, but are not limited to, moisturizer emulsions (e.g., lotion), conditioner emulsions, cleanser emulsions, and any combinations thereof.

[0026] In any embodiment, the molecule in which the sensor is configured to detect may include an allergen, a trace molecule of an allergen, a pathogen, a toxin, or a combination of any two or more thereof. [0027] As used herein “allergen” refers to both allergy and intolerant inducing substances. A true allergy causes an immune system reaction that affects numerous organs in the body and can cause a range of symptoms. In some cases, an allergic reaction can be severe or life-threatening. In comparison, intolerance symptoms are generally less serious and often limited to digestive problems. Nonlimiting examples of intolerances include absence of an enzyme needed to fully digest a consumable (e.g., food or drink), irritable bowel syndrome, sensitivity to an additive, recurring stress or psychological factors, and celiac disease. An example of an absence of an enzyme is lactose intolerance. Irritable bowel syndrome is a chronic condition that may cause cramping, constipation, and/or diarrhea. An example of sensitivity to an additive are sulfites commonly used to preserve food and drinks. Celiac disease has some features of a true food allergy because it involves the immune system, however, symptoms are mostly gastrointestinal, and people with celiac disease are not at risk of anaphylaxis.

[0028] Allergens may include, but are not limited to, animal products, grains (e.g., gluten), vegetables, fruits, dairy products, fish, beverages, legumes, chocolates, synthetic food chemicals (e.g., monosodium glutamate (MSG), artificial sugars such as aspartame), and any combinations of two or more thereof. In one example, an allergen may include a food protein. In any embodiment, the allergen or the trace molecule may be a peanut allergen, tree nut allergen, milk allergen, egg allergen, wheat allergen, soy allergen, meat allergen, fish allergen, shellfish allergen, coconut allergen, or a combination of two or more thereof. In any embodiment, the allergen or the trace molecule may be a nut allergen listed in Table 1. In any embodiment, the allergen or the trace molecule may be a tree nut allergen (e.g., almond, almond paste, or a combination thereof). In any embodiment, the allergen or the trace molecule may be a soy allergen. In any embodiment, the allergen or the trace molecule may include a flavonoid, amygdalin, or a combination thereof. In any embodiment, the flavonoid may include an isoflavonoid, neoflavonoid, or derivatives thereof. In any embodiment, the isoflavonoid or derivative thereof may include isoflavones, isoflavonones, isoflavans, pterocarpans, rotenoids, or combinations of two or more thereof. In any embodiment, the allergen or the trace molecule may include amygdalin, apigenin-6- arabinoside-8-glucoside,apigenin-6-glucoside-8-arabinoside, arachin, biochanin A, catechin gallate, crysoeriol, cyanocobalamin, daidzein, daidzin,5-5'-dehydrodiferulic acid, 5-8'- dehydrodiferulic acid, 5,7-dihydroxychromone, 5,7, dimethoxyisoflavone, ferulic acid, galactose, genistein, genistin, 3 -hydroxybiochanin A, isochlorogenic acid, isoferulic acid, juglone, lactose, lariciresinol, medioresinol, procyanidin B2, procyanidin Cl, resveratrol, resveratrol 3-glucoside, secoisolariciresinol, syringaresinol, syringic acid, trans-sinapic acid.

[0029] As used herein a “trace molecule of an allergen” refers to molecules that are suitable for detecting the presence of an allergen but may not necessarily be allergens themselves. In any embodiment, the trace molecule of the allergen may be an organic molecule or a salt thereof. For example, the trace molecule may be the allergen itself, epitope of an allergen (i.e., the part of an antigen molecule to which an antibody attaches itself), molecule that is commonly present with an allergen, a subunit of an allergen, a derivative of an allergen, or a combination of two or more thereof including a polypeptide, protein, epitope, aptamer, or a combination of any two or more thereof. In some embodiments, the organic molecule may include at least one protein. In some embodiments, the organic molecule may include at least two different proteins. In some embodiments, the organic molecule may include at least one epitope. In some embodiments, the organic molecule may include at least two different epitopes. In some embodiments, the organic molecule may include at least one protein and at least one epitope. In some embodiments, the organic molecule may be selected from lactose, galactose, amygdalin, juglone, biochanin A, resveratrol daidzein, daidzin, genistein, genistin, and a combination of any two or more thereof. In any embodiment, the organic molecule may not include cortisol, an amino acid, theophylline, and/or chlorpyrifos.

[0030] Due to the high frequency of severity of peanut allergies, a peanut-related allergen is used in an exemplary fashion in this disclosure. It is contemplated that other allergens may replace the peanut-related allergen in the example, embodiment, implementation or other aspect of the disclosure. One way to test for the presence of a peanut-related allergen is to test for a peanut protein allergen or an epitope thereof. Examples of a peanut protein include, but are not limited to, arachis hypogaea allergen 1 (Ara Hl), arachis hypogaea allergen 2 (Ara H2), arachis hypogaea allergen 3 (Ara H3), arachis hypogaea allergen 4 (Ara H4), arachis hypogaea allergen 5 (Ara H5), arachis hypogaea allergen 6 (Ara H6), arachis hypogaea allergen 7 (Ara H7), arachis hypogaea allergen 8 (Ara H8), arachis hypogaea allergen 9 (Ara H9), arachis hypogaea allergen 10 (Ara H10), arachis hypogaea allergen 11 (Ara Hl 1), arachis hypogaea allergen 12 (Ara H12), arachis hypogaea allergen 13 (Ara H13), arachis hypogaea allergen 14 (Ara H14), arachis hypogaea allergen 15 (Ara Hl 5), arachis hypogaea allergen 16 (Ara Hl 6), arachis hypogaea allergen 17 (Ara Hl 7), peanut agglutinin (PNA), and combinations of any two or more thereof. In some embodiments, the device may be configured to detect any of peanut proteins provided herein or an epitope thereof including Ara Hl, Ara H2, Ara H3/H4, Ara H6 epitope, or a combination of any two or more thereof. Further specific examples of allergen epitopes are listed in Table 1 below.

Table 1 : Exemplary list of allergen epitopes

[0031] Pathogens may include, but are not limited to, a bacterium, virus, or other microorganism that can cause disease and/or illness. In any embodiment, the pathogen may be a food pathogen and/or a clinical pathogen. Exemplary food pathogens include, but are not limited to, Campylobacter, Cyclospora, Clostridium botulinum, Escherichia coli, Listeria, Salmonella, Staphylococcus aureus, Shigella, Toxoplasma gondii, Vibrio vulnificus, Norovirus, Hepatitis A, or a combination of any two or more thereof. Exemplary clinical pathogens include, but are not limited to, Candida, Chlamydia trachomatis, Neisseria gonorrhoeae, Methicillin-resistant Staphylococcus aureus, Mycobacterium tuberculosis, human papillomavirus (HPV), Hepatitis B, Hepatitis C, Hepatitis D, Hepatitis E, human immunodeficiency virus, influenza, or a combination of any two or more thereof.

[0032] Toxins may include, but are not limited to, herbicides, pesticides, drugs of abuse, or a combination of any two or more thereof. As used herein, herbicides refers to substances that are toxic to plants and commonly used to destroy unwanted vegetation. As used herein, herbicides refers to substances that are toxic to insects or other organisms harmful to cultivated plants or to animals. Exemplary herbicides and/or pesticides include atrazine, azinphos-methyl, bentazone, carbaryl, carbofuran, chlorpyrifos methyl, chlorsulfuron, cyhexatin, diazinon, dimethoate, fenobucarb, glyphosate, hydrazine, imidacloprid, lindane, methyl parathion, paraquat, parathion, permethrin, pirimicard, sulfentrazone, or a combination of any two or more thereof. As used herein, drugs of abuse refers to illegal drugs as well as prescription or over-the-counter drugs that are used for purposes other than those for which they are meant to be used, or in excessive amounts. Exemplary drugs of abuse include an amphetamine or a metabolite thereof (e.g., methamphetamine, 3,4-methylenedioxyamphetamine (MDA), phentermine, ephedrine, and/or pseudoephedrine), cocaine or a metabolite thereof (e.g., benzoylecgonine), a benzodiazepine or a metabolite thereof (e.g., diazepam, temazepam, chlordiazepoxide, nordiazepam, oxazepam, a-hydroxyalprazolam, a-hydroxytriazolam, 7-aminoclonazepam, 7-aminoflunitrazepam, and/or hydroxyethyl-flurazepam), a barbiturate or a metabolite thereof (e.g., mephobarbital. Phenobarbital, butalbital, amobarbital, pentobarbital, and/or secobarbital), a dissociative drug or a metabolite thereof (e.g., ketamine and/or norketamine), an opioid or a metabolite thereof (e.g., fentanyl, norfentanyl, methadone, 2- ethylidene-l,5-dimethyl-3,3- diphenylpyrrolidine (EDDP), buprenorphine, norbuprenorphine, diacetylmorphine (heroin), 6-monoacetylmorphine (6-MAM), morphine, codeine, hydrocodone, hydrocodone, norhydrocodone, dihydrocodeine, hydromorphone, oxycodone, oxymorphone, naloxone, noroxymorphone, and/or noroxycodone), or a combination of any two or more thereof.

[0033] In any embodiment, the molecule may be in salt form (i.e., ionic molecule). In any embodiment, the one or more polymerized monomers may include a monomer of formula I, IV, V, VI, VII, and/or IX having a hydrogen acceptor or hydrogen donor functional group (e.g., -CO2H, OH, and/or NH2). In any embodiment, the MIP and NIP may be at a pH such that the one or more polymerized monomers are capable of ionic interactions (e.g., the pH is at a pH such that the hydrogen acceptor or hydrogen donor functional group is in ionic form). For example, the pH may be at 6 such that the amino group of the monomer of formula IX is positively charged and the molecule is negatively charged resulting in interactions between the positively charged polymerized monomers and the negatively charged molecule.

[0034] In any embodiment, the molecule may be in salt form and the one or more polymerized monomers may include a monomer of formula V. In any embodiment, the monomer of formula V may be doped with F; Br", Cl", NCh’, CICU-, SCU^2-, PCU^3-, or a combination of any two or more thereof.

[0035] In some embodiments, the molecule may have one or more aromatic groups and the one or more polymerized monomers may be any disclosed herein having one or more aromatic groups (e.g., 3, 4-ethylenedi oxythiophene or a compound of formula I, II, V, VI, VII, VIII, or IX).

[0036] In any embodiment, the one or more polymerized monomers may include a monomer of formula I, II, III, IV, V, VI, VII, VIII, or IX. In some embodiments, the one or more polymerized monomers may be a homopolymers (i.e., one of the monomers of I-IX). In other embodiments, the one or more polymerized monomers may be a copolymer (z.e., at least two of the monomers of I-IX or one of the monomers of I-IX plus at least one additional polymerized monomer). In any embodiment, the polymer of the MIP and NIP include the same polymerized monomers.

[0037] In any embodiment, the one or more polymerized monomers may include a monomer of formula I. In some embodiments, R¹ is NHz and R² and R³ are H. In other embodiments, R¹, R², and R³ are H.

[0038] In any embodiment, the one or more polymerized monomers may include a monomer of formula II. In some embodiments, R⁴ is H; A, B, E, and G are CH; and D is N.

[0039] In any embodiment, the one or more polymerized monomers may include a monomer of formula III. In some embodiments,

In any embodiment, the one or more polymerized monomers may include a second monomer. In any embodiment, the one or more polymerized monomers may include a first monomer of formula III and a second monomer of formula III. In some embodiments, R⁵ is

H and R⁶ is H in the first monomer

the second monomer.

[0040] In any embodiment, the one or more polymerized monomers may include a monomer of formula IV. In some embodiments, R⁷ is CH3, R⁸ is OH, and m is 2.

[0041] In any embodiment, the one or more polymerized monomers may include a monomer of formula V. In some embodiments, J is N and R⁹ is H. In some embodiments, J is S and R⁹ is absent. [0042] In any embodiment, the one or more polymerized monomers may include a monomer of formula VI. In some embodiments, R¹⁰ is H and R¹¹ is H. In some embodiments, R¹⁰ is H and R¹¹ is CH2-CH2-NH2.

[0043] In any embodiment, the one or more polymerized monomers may include a monomer of formula VII. In some embodiments, R¹², R¹³, R¹⁹, and R²⁰ are H and R¹⁴ is NH2. In some embodiments, R¹² and R²⁰ are H and R¹³, R¹⁴, and R¹⁹ are OH.

[0044] In any embodiment, the one or more polymerized monomers may include a monomer of formula VIII. In some embodiments, R¹⁵ is C1-C3 alkyl. In some embodiments, R¹⁵ is CH2-CH3. In some embodiments, R¹⁵ is CH3.

[0045] In any embodiment, the one or more polymerized monomers may include 3,4- ethy 1 enedi oxy thi ophene .

[0046] In any embodiment, the one or more polymerized monomers may include a monomer of formula IX. In some embodiments, R¹⁶ is NH2 and R¹⁷ and R¹⁸ are each H. In some embodiments, R¹⁶, R¹⁷, and R¹⁸ are each H. In some embodiments, R¹⁶ is SH and R¹⁷ and R¹⁸ are each H.

[0047] In some embodiments, the molecule may have a molecular weight less than about 1000 g/mol (e.g., the molecule may be a small molecule and/or toxin). In other embodiments, the molecule may be a pathogen having a molecular weight greater than about 1000 g/mol (e.g., E. Coli or Salmonella having a molecular weight of 54.6 kDa and 29.5 kDa, respectively). Any of the one or more polymerized monomers disclosed herein may be used to detect the presence of a small molecule. Preferably, the one or more polymerized monomers disclosed herein that are hydrophobic may be used for detecting a molecule that is hydrophobic and the one or more polymerized monomers disclosed herein that are hydrophilic may be used for detecting a molecule that is hydrophilic. For example, if the molecule is more hydrophilic, then one or more polymerized monomers would be those that are more hydrophilic such that ionic and/or electrostatic interactions are exhibited. For highly hydrophobic molecules, the one or more polymerized monomers would be those that are more hydrophobic (e.g., the one or more polymerized monomers could be any of those disclosed except HEMA, dopamine, or acrylamide). For example, in some embodiments, the molecule is hydrophobic and the one or more polymerized monomers may include a monomer of formula I, II, V, VI, VII, or IX and be hydrophobic.

[0048] In some embodiments, the polymer of the MIP and/or NIP may exclude polymerized monomers selected from 3 -aminophenyl boronic acid, 4-aminophenyl boronic acid, 2-hydroxyphenyl boronic acid, 3-hydroxyphenyl boronic acid, 4-hydroxyphenol boronic acid, pyrrole, polyaniline, thiophene, 3, 4-ethylenedi oxythiophene, phenylene diamine, phenyl boronic acid, p-aminothiophenol, aminophenol, p-phenyl phenylenediamine, o-toluidine, and combinations of any two or more thereof.

[0049] In any embodiment, the one or more polymerized monomers further comprises a cross-linker. In any embodiment, the cross-linker may include ethylene glycol dimethacrylate (EGDMA), trimethyl tripropane triacrylate (TMPTA), glycerol, glutaraldehyde, dimethacrylate, or a combination of any two or more thereof. In any embodiment, the cross-linker(s) may be added during or before polymerization of the one or more polymerized monomers to form the MIP and NIP. EGDMA and glycerol dimethacrylate are difunctional and TMPTA is trifunctional. Each of the functional groups may be reacted with separate polymer chains such that the chains are cross-linked. Other cross-linkers known by those of ordinary skill in the art for surface imprinting, including for conventional surface imprinting, may also be used.

[0050] In any embodiment, the electropolymerized MIP and the molecule may have a complexation energy of about -50 kJ/mol to about -1500 kJ/mol overall (i.e., per MIP). In any embodiment, the electropolymerized MIP and the molecule may have a complexation energy of about -70 kJ/mol to about -1250 kJ/mol overall. In any embodiment, the electropolymerized MIP and the molecule may have a complexation energy of about -100 kJ/mol to about -1000 kJ/mol overall. In any embodiment, the electropolymerized MIP and the molecule may have a complexation energy of about -60 kJ/mol to about -1000 kJ/mol overall including about -80 kJ/mol to about -900 kJ/mol, about -100 kJ/mol to about -800 kJ/mol, about -125 kJ/mol to about -750 kJ/mol, about -150 kJ/mol to about -700 kJ/mol, about -175 kJ/mol to about -650 kJ/mol, about -200 kJ/mol to about -600 kJ/mol, about -250 kJ/mol to about -600 kJ/mol, about -275 kJ/mol to about -550 kJ/mol, about -300 kJ/mol to about -500 kJ/mol, about -100 kJ/mol to about -350 kJ/mol, or about -150 kJ/mol to about -300 kJ/mol. In any embodiment, the electropolymerized MIP and the molecule may have a complexation energy of less than 0 kJ/mol per binding site. In any embodiment, the electropolymerized MIP and the molecule may have a complexation energy of about -10 kJ/mol to about -150 kJ/mol per binding site including about -20 kJ/mol to about -125 kJ/mol per binding site. In any embodiment, the electropolymerized MIP and the molecule may have a complexation energy of about -30 kJ/mol to about -100 kJ/mol per binding site including about -50 kJ/mol to about -100 kJ/mol, about -60 kJ/mol to about -100 kJ/mol, about -70 kJ/mol to about -100 kJ/mol, about -40 kJ/mol to about -90 kJ/mol, about -40 kJ/mol to about -80 kJ/mol, or about -40 kJ/mol to about -70 kJ/mol. In any embodiment, the electropolymerized MIP and the molecule may have a complexation energy of about -10 kJ/mol to about -70 kJ/mol per binding site including about -15 kJ/mol to about -60 kJ/mol or about -20 kJ/mol to about -50 kJ/mol. In any embodiment, the electropolymerized MIP and the molecule may have a complexation energy of about -75 kJ/mol to about -150 kJ/mol per binding site including about -80 kJ/mol to about -150 kJ/mol, about -90 kJ/mol to about -150 kJ/mol, about -100 kJ/mol to about -150 kJ/mol, about -110 kJ/mol to about -150 kJ/mol, about -120 kJ/mol to about -150 kJ/mol, or about -130 kJ/mol to about -150 kJ/mol.

[0051] In any embodiment, the one or more polymerized monomers may be determined by computationally calculating the complexation energy of the one or more polymerized monomers and the molecule.

[0052] Nonlimiting exemplary complexation energy ranges for the one or more polymerized monomers and molecules disclosed herein are provided in Tables 2-6.

Table 2: Complexation energy of exemplary pairings with peanut epitopes NNPFYFPSR, SFNLDEGHALR, NTLEAAFNAEFNEIR, VLLEENAGGEQEER, DLAFPGSGEQVEK, GTGNLELVAVR, QSQLER, CMCEALQQIMENQSDR, RQQWELQGDR, DPYSPS, KRELRNLPQQ, SPDIYNPQAGSLK, SQSENFEYVAFK, RPFYSNAPQEIFIQQGR, WLGLSAEYGNLYR, YDYSIR, or KRELRMLPQQ.

Table 3: Complexation energy of exemplary pairings with egg epitopes AAFGAEVDCSRFPNATD, RFPNATDKEGKDVLV, SIEFGTNISKEHDG, PMNCSSYANT, ITKPNDVYSFSLA, DEDTQAMPFRVTEQ, SGTMSMLVLLPDE, or PDEVSGLEQLESIIN.

Table 4: Complexation energy of exemplary pairings with milk epitopes VKKILDKVGINY, LKDLKGYGGV, RYLGYLEQLLRLKK, ELAYFYPELFRQF, NEINQFYQKFPQYLQYL, KPWIQPKTKVIPY, NAVPITPTLNREQLS, VVVPPFLQPEVMGV, HLPLPLLQSWMH, SFMAIPPKKNQDKTE, PSYGLNYYQQKPV.

Table 5: Complexation energy of exemplary pairings with fish epitope AAGSFDHKKFFKACGLSGKSTDEVK.

Table 6: Complexation energy of exemplary pairings with wheat epitopes VRVPVPQLQP, QEQVPLVQQQ, VQQQQFPGQQ, LALQTLPAMC, QPQQPFPQ, QQSGQGQ, HQQQPIQQQP, or QSRYEAIRAI.

[0053] In any embodiment, the electropolymerized MIP film may further include a tracer. When a tracer is present, following imprinting and removal of the molecule in the MIP, the tracer can permeate the polymer surface to provide a current signal via the tracer’s oxidation or reduction. When the molecule is present and rebound to the MIP, the current signal of the MIP will decrease by closing the pores/cavities and therefore causing a concentrationdependent decrease in the permeation of the tracer.

[0054] In any embodiment, the tracer may be selected from the group consisting of potassium ferricyanide/ferrocyanide, hexaammineruthenium (II)/(III) chloride, ferrocenecarboxylic acid (II)/(III), hydroquinone, ferrocene, methylene blue, iridium (II)/(III) chloride, ascorbic acid, dopamine, and a combination of any two or more thereof. In any embodiment, the molecules disclosed herein for detecting may be the tracer. In any embodiment, the electropolymerized MIP has about 1 nmol to about 500 mmol of the tracer. In any embodiment, the electropolymerized MIP has about 1 pmol to about 500 mmol of the tracer. In any embodiment, the electropolymerized MIP has about 1 pmol to about 1 mmol of the tracer. In any embodiment, the electropolymerized MIP has about 1 mmol to about 1 mmol of the tracer.

[0055] In any embodiment, the electropolymerized MIP film and/or electropolymerized NIP film may include more than one layer of the polymer. In any embodiment, the electropolymerized MIP film and/or electropolymerized NIP film may include two or more layers of the polymer.

[0056] In any embodiment, the electropolymerized MIP film and/or electropolymerized NIP film may have a thickness of about 2 nm to about 100 nm, about 2 nm to about 75 nm, about 2 nm to about 50 nm, about 2 nm to about 40 nm, about 2 nm to about 30 nm, or about 2 nm to about 20 nm.

[0057] In any embodiment, the device may further include one or more electrochemical chips. In some embodiments, the device may include one electrochemical chip comprising the electropolymerized MIP film and the electropolymerized NIP film. In some embodiments, the device may include a first electrochemical chip, wherein the first electrochemical chip comprises the electropolymerized MIP film and/or a second electrochemical chip, wherein the second electrochemical chip comprises the electropolymerized NIP film. In an embodiment, the molecule may be immobilized (e.g., covalently) to the electrochemical chip prior to forming the MIP film.

[0058] In any embodiment, the first electrochemical chip may include a working electrode, a counter electrode, and a reference electrode. In any embodiment, the second electrochemical chip may include a working electrode, a counter electrode, and reference electrode. In any embodiment, the working, counter, and/or reference electrode(s) may include carbon. In any embodiment, the working and/or counter electrodes may include glassy carbon, carbon nanotubes, graphene, gold, platinum, silver, chromium, graphite, carbon black, or a combination of two or more thereof. In any embodiment, the reference electrode material may include be silver (e.g., silver chloride), calomel electrode, standard hydrogen electrode, normal hydrogen electrode, palladium hydrogen electrode, or a combination of two or more thereof. In any embodiment, the working electrode may have a diameter of about 0.1 mm to about 5 mm. In some embodiments, the device may include a single electrochemical chip that includes both the electropolymerized MIP film and the associated electrode and the electropolymerized NIP film and the associated electrode. The single electrochemical chip may be produced by the same methods and components as the first and second electrochemical chips described herein. [0059] In any embodiment, the surface of the working electrode, the counter electrode, and/or the reference electrode may be modified. In any embodiment, the surface of the working electrode may be modified. Modification includes the addition of a conductor(s) and/or a semiconductor(s) to the electrode surface. In any embodiment, the conductor(s) and/or semiconductor s) may include carbon materials, conductive polymers, nanoparticles, or a combination of two or more thereof. Carbon materials may include carbon nanotubes (e.g., single walled and/or multiwalled), fullurenes, graphene, reduced graphene oxide, or combinations of two or more thereof. Conductive polymers may include polyaniline, polypyrrole, polythiophene, poly(3,4-ethylenedi oxy thiophene), poly(o-toluidine), polyacetylene, polyphenylenes, polypyrenes, polyazulenes, polynaphthalenes, polycarbazole, polyindoles, polyazepines, poly(/?-phenylene sulfides), polyfluorenes, or combinations of two or more thereof. Nanoparticles may include spherical nanoparticles, nanowires, nanorods, nanourchins, nanoshells, nanocubes, nanoplates, nanoribbions, or combinations of two or more thereof. Nanoparticles may include metal(s) such as gold, silver, platinum, chromium, palladium, or combinations of two or more thereof. Nonlimiting modes of adding the conductor(s) and/or the semiconductor(s) to the electrode surface include depositing the modifying material by way of physical deposition (e.g., drop cast, spin cast, or screen printed) and/or electrochemical deposition (e.g., electropolymerization of a polymer or reduction of a carbon material). In any embodiment, the surface modification may improve the mechanical, chemical, and/or electronic interface.

[0060] In any embodiment, the device may further include a circuit board (e.g., printed circuit board) comprising the first electrochemical chip and the second electrochemical chip. In any embodiment, the circuit board may include the first electrochemical chip and the second electrochemical chip. In any embodiment, the electrochemical chips may be produced in any known manner including screen printing, inkjet printing, vapor deposition, lithography, or subtractive methods. In any embodiment, the electrochemical chips may be produced using gold, carbon screen-printed electrodes, or gold electrodes prepared by electrodeposition. In any embodiment, the circuit board may be width and thickness to fit a standard interface (e.g., SD, MicroSD, USB, or USB-C). Non-limiting examples of PCB material include FR4, bakelite, glass, plastic, rubber, cellulose, and the like. In any embodiment, the circuit board may be about 1 cm² in area and have an interdigit spacing of 300 pm. Any circuit board known to those of skill in the art may be used in the present technology. Non-limiting illustrative examples can be found in PCT/US2019/058833 (herein incorporated by reference).

[0061] In any embodiment, the device may further include a substrate. In any embodiment, the circuit board that includes the electrochemical chips may further include a substrate. In any embodiment, the substrate may have been or may be exposed to the sample. In any embodiment, the substrate may include glass, plastic, paper, quartz, alumina, mica, silicon, a III-IV semiconductor compound, or combinations of two or more thereof. In any embodiment, the substrate may include copper on a PCB material in an interdigitated pattern. In some embodiments, the copper may be laminated on one or both sides of the PCB material.

[0062] In any embodiment, the device may include a body that includes a capsule that encapsulates a solvent and chamber configured to receive the tangible good sample. In any embodiment, the device may further include a substrate with a tangible good sample on the surface configured for insertion into the chamber. In any embodiment, the chamber may also provide an area for mixing the solvent with the tangible good sample. In any embodiment, the body may at least partially surround the sensor, capsule, and chamber. In any embodiment, the body may at least partially surround the sensor, capsule, chamber, and substrate. In any of the above embodiments, the device further comprises a recess configured to house the sensor. In any embodiment, the body may be a multi-use body. In any embodiment, the body may be a one-time use body. In any embodiment, the body may be disposable, recyclable, and/or compostable. In any embodiment, the body may be recyclable. A typical disposable body may contain multiple sensors, including one or more first electropolymerized chips and/or one or more second electropolymerized chips. In any embodiment, the sensor may include one or more additional electropolymerized chips that include an MIP of another molecule different from the molecule of the first electropolymerized chip. [0063] In any embodiment, after exposing the tangible good sample to the substrate, the substrate may be exposed to the sensor. Exposure may be direct or the substrate may first be exposed to a liquid solvent that in turn solubilizes, extracts, mixes, and/or encourage selective binding of the potential molecule from the tangible good sample. Alternatively, the solvent may be used to reduce the solubility of a molecule, altering the equilibrium between being dissolved in the solvent and bound to the MIP. In any embodiment, the solvent(s) may be stored in compartments, capsules, or pouches inside a disposable unit. In any embodiment, the body may include a capsule that encapsulates the liquid solvent. In any embodiment, the solvent may include water, aqueous buffer, an electrolyte solution, an organic solvent (e.g., ethanol), or a combination of two or more thereof. In any embodiment, the solvent may include an aqueous buffer. In any embodiment, the aqueous buffer may include a mild alkaline buffer solution (pH -9-11 carbonate/ bicarbonate). In any embodiment, the solvent may include an electrolyte solution (e.g., potassium chloride solution). In any embodiment, the device may include a chamber for mixing the solvent with the tangible good sample. In any embodiment, the chamber may be configured to exhibit mixing the solvent with the tangible good sample through agitation (e.g., physical agitation such as shaking, stirring, and/or grinding) to produce a substantially homogenized mixture. In any embodiment, release of the solvent may prevent reopening of the body and/or prevent release of the solvent and tangible good sample from the body. In any embodiment, the sample may be incubated with the solvent (e.g., from about 1 second to 30 minutes, from about 2 seconds to 10 minutes, or from about 5 seconds to 5 minutes). Incubation to allow for molecule binding and electrochemical probing may be separate events, or they may happen simultaneously. Under simultaneous conditions, the solvent may further include an appropriate redox probe electrolyte (e.g., K4Fe(CN)e / K3Fe(CN)e and/or Ru(NH3)eC13 / Ru(NH3)6Ch). Under separate events, after incubation the sensor may be moved to an appropriate redox probe electrolyte (e.g., K4Fe(CN)e / K3Fe(CN)e and/or Ru(NH3)eC13 / Ru(NH3)6Ch). In particular, when the molecule is a non-redox active molecules (e.g., epitopes and proteins), testing for presence or absence of the molecule includes the solvent containing an appropriate redox probe electrolyte (i.e., simultaneous events) or after incubation moving the sensor to an appropriate redox probe electrolyte (i.e., separate events). When the molecule is a redox active molecule including an appropriate redox probe electrolyte is optional. In any embodiment, the sample may or may not undergo purification steps such as filtration or dialysis.

[0064] In any of the above embodiments, the mixture of the tangible good sample and solvent may be exposed to the electropolymerized MIP film and electropolymerized NIP film. In any of the above embodiments, the mixture of the tangible good sample and solvent may be exposed to the electropolymerized MIP film and the associated electrode and the electropolymerized NIP film and the associated electrode. In any embodiment, the mixture of the tangible good sample and solvent may be exposed to the electropolymerized MIP film and electropolymerized NIP film through puncturing a barrier. After the electropolymerized MIP film and electropolymerized NIP film are exposed to the mixture of the tangible good sample and solvent, the printed circuit board (comprising the electropolymerized MIP film and electropolymerized NIP film) terminating in a connector (e.g., MicroSD) may be inserted into the Amulet processing device. The printed circuit board (including the electropolymerized MIP film and electropolymerized NIP film) and the connector may be physically arranged in any way known to those of ordinary skill in the art such that the mixture of the tangible good sample and solvent may be exposed to the electropolymerized MIP film and electropolymerized NIP film and the connector is available for insertion into the Amulet processing device.

[0065] In any embodiment, the sensor may be stored in a dry compartment within the disposable or in a compartment containing a solvent. In any embodiment, additional chemicals may also be mixed with the sample to modulate the solubility of the tracer molecule. Such chemicals include buffers, salts, and surfactants. In any embodiment, the chemicals may be stored in the same chamber as the sensor or in a separate chamber.

[0066] In any embodiment, the processing device is configured to communicatively couple to the sensor. In some embodiments, the processing device comprises circuitry configured to determine presence of the molecule. In another embodiment, to determine presence of the molecule the processing device is configured to compare an electric current of the MIP to the electric current of the NIP. In further embodiments, the processing device is configured to determine an electric current difference between an electric current of the MIP and an electric current of the NIP; compare the electric current difference to a threshold difference. In further embodiments, the processing device is configured to determine that the molecule is present when the electric current difference is greater than the threshold difference. In further embodiments, the processing device determines that the molecule is present when the electric current of the MIP is greater than the electric current of the NIP. In yet further embodiments, the processing device determines that the molecule is present when the electric current of the MIP is less than the electric current of the NIP.

[0067] In any embodiment, the device may further include a re-usable reader or a processing device (the “Amulet”). Commonly, the processing device may be configured to communicatively couple to the sensor and may be configured to determine an electric current and/or potential difference between the electropolymerized MIP film and the electropolymerized NIP film (e.g., may include multimeter/ potentiostat/ microprocessor/ physical memory). In any of the above embodiments, the processing device may determine the presence of the molecule when the electric current and/or potential difference of the electropolymerized MIP film is greater than the electric current and/or potential difference of the electropolymerized NIP film. In any of the above embodiments, the processing device may determine the presence of the molecule when the electric current of the electropolymerized MIP film is greater than the electric current of the electropolymerized NIP film. In any of the above embodiments, the processing device may determine the presence of the molecule when the electric potential difference of the electropolymerized MIP film is greater than the electric potential difference of the electropolymerized NIP film. In any of the above embodiments, the processing device may determine the presence of the molecule when the electric current and/or potential difference of the electropolymerized MIP film is lower than the electric current and/or potential difference of the electropolymerized NIP film. In any of the above embodiments, the processing device may determine the presence of the molecule when the electric current of the electropolymerized MIP film is lower than the electric current of the electropolymerized NIP film. In any of the above embodiments, the processing device may determine the presence of the molecule when the electric potential difference of the electropolymerized MIP film is lower than the electric potential difference of the electropolymerized NIP film. In any embodiment, the electric current and/or potential difference may be determined by cyclic voltammetry (CV), linear sweep voltammetry, square wave voltammetry, differential pulse voltammetry, amperometry, or a combination of two or more thereof. In any embodiment, the electric current and/or potential difference may be determined by cyclic voltammetry (CV). In any embodiment, the processing device may determine the presence of the molecule when the resistance of the MIP film is lower than the resistance of the NIP film. In any embodiment, the processing device may determine the presence of the molecule when the resistance of the MIP film is higher than the resistance of the NIP film.

[0068] The relative measurements of each chip are used to determine whether the molecule is present. Readings for each chip may be taken once, multiple times, or continuously. In some embodiments, the device may further comprise a processing device, wherein the processing device is configured to communicatively couple to the sensor, wherein the processing device is configured to determine an electric current difference between an electric current of the MIP film and an electric current of the NIP film. In further embodiments, the processing device determines the presence of the first molecule when the electric current of the electric current of the MIP film is greater than the electric current of the NIP film. In another embodiment, the processing device determines the presence of the first molecule when the electric current of the electric current of the MIP film is less than the electric current of the NIP film.

[0069] In any embodiment, the processing device communicatively couples to the sensor via a plurality of contacts of the sensor and via a plurality of contacts of the processing device. In further embodiments, the processing device is a wearable. In another embodiment, the processing device communicatively couples to the sensor via a wireless signal. In further embodiments, the wireless signal comprises a radio and/or infrared frequency signal. In yet further embodiments, the processing device is a computer, telephone, watch, and/or mobile device.

[0070] In another aspect, the present technology provides a method of making the detection device described herein. MIPs and NIPs may be manufactured by methods known to those of skill in the art including those provided in U.S. Patent No. 9,846,137, which is herein incorporated by reference. In any embodiment, the method may include providing a conductive electrode, depositing a polymer in the presence of the molecule by electropolymerization to form the electropolymerized MIP film, and depositing the polymer in the absence of the molecule by electropolymerization to form the electropolymerized NIP film. In any embodiment, the depositing the polymer on a first electrochemical chip in the presence of the molecule provides the first electropolymerized chip and/or the depositing the polymer on a second electrochemical chip in the absence of the molecule provides the second electropolymerized chip. The polymer may be any polymer described herein. The molecule is in a tangible good and may be any molecule described herein. In any embodiment, the first and second electropolymerized chips may take any reasonable size and pattern for measuring the electric current of the MIP and NIP films. In any embodiment, the electropolymerized chips may be used for a 2-point electric current measurement, a 4- point electric current measurement, or more complex electrochemical measurements as described herein (e.g., CV, linear sweep voltammetry, square wave voltammetry, etc.).

[0071] The methods include forming the polymerized film on the sensor that has been modified with the molecule, such that after polymerization, the template molecule is amenable to being extracted from the film leaving behind a cavity that is specific to the molecule that was extracted. The molecules may be either near the surface of the polymer film, or attached to the surface of the sensor in a substantially perpendicular arrangement to the surface of the sensor, such that once the polymer film is formed, the molecule may be removed or cleaved from its anchor to the surface, thus leaving the cavity. Then, during operation, when a target molecule of the same, or substantially the same, identity as the molecule that was removed is present, it may fit into and bind into the cavity with the sensor positively identifying its presence in the sample. The removal of the molecule can include cleaving it from the surface (i.e. from the sensor substrate) or by cleaning an bond within the molecular structure extending to the surface (i.e. a cleavable S-S bond within an epitope that may be used as the template molecule). Thus, the thickness of the electropolymerized polymer film is controlled such that it is of a sufficient thickness, but not so thick as to prevent extraction of the template molecule. In some embodiments, this may be from 1 nm to 100 nm, from 1 nm to 50 nm, or from 2 nm to 10 nm, or from 2 nm to 4 nm. In other embodiments, this may be from 1 nm to 10 pm, from 100 nm to 10 pm, or from 100 nm to 5 pm, or from 1 pm to 5 pm.

[0072] In general, MIP films are synthesized by combining functional monomers/polymers with a molecule to provide a pre-polymerization solution, submerging an electrochemical chip in the pre-polymerization solution, and connecting the chip to a potentiostat. In any embodiment, the pre-polymerization solution may include a solvent (e. ., water, ethanol acetonitrile, acetone, tetrahydrofuran, dimethylsulfoxide, dimethylformamide, N-methylpyrollidone, N,N-dimethylacetamide, or a combination of two or more thereof). In any embodiment, the pre-polymerization solution may include a buffer (e.g., acetate buffers, carbonate buffers, citrate buffers, phosphate buffers, or a combination of two or more thereof). In any embodiment, the pre- polymerization solution may include an electrolyte (e.g., FeCh, KC1, tetraalkylammonium salts, LiClOr, LiTFMS, or a combination of two or more thereof. In any embodiment, the molecule may have a concentration ranging from nanomolar to millimolar. In any embodiment, the pre-polymerization solution may be prepared at room temperature, but may be performed at higher or lower temperatures. In any embodiment, the pre-polymerization solution is prepared at least 5 minutes to 1 hour prior to electropolymerization to allow enough time for complexation between the monomer/polymer and the molecule.

[0073] In any embodiment, the potential of the working electrode may be cycled through a range of voltages which causes a film to polymerize onto the electrode surface. For example, potentiostat cycles may range from about -2 V to about 2 V (including 0 to about 1 V), about 1-100 times (including about 10-30 times), at various rates such as about 1 mV/s to about 1 V/s (including about 40 mV/s to about 60 mV/s). In any embodiment, a single chip may be polymerized at a time, or multiple chips may be connected in parallel and coated as a batch.

[0074] After a series of cycles, the molecule may be removed from the polymer. In any embodiment, removal of the molecule from the MIP film may be achieved by using a solvent, surfactant, buffer, electrochemistry, or a combination thereof. For example, the molecule may be removed by rinsing it away, over-oxidizing, or electrochemical stripping (anodic or cathodic desorption by applying and holding the electrode at either positive or negative potential, respectively). For example, some molecules can be removed from the electrode by holding the electrode at -1.1V vs Ag/AgCl for 30 sec. Removal of the molecule leaves behind an MIP film with empty molecular cavities. In any embodiment, the solvent may be any solvent capable of dissolving the molecule but not the polymer film (e. ., methanol, ethanol, acetonitrile, THF, DMF, DMSO, etc ). In any embodiment, an appropriate surfactant (anionic, cationic, or neutral) may be added. Anionic surfactants include, but are not limited to, alkylbenzene sulfonates, fatty acid soaps, dialkyl sulfosuccinate, alkyl ether sulfates, sulfated alkanolamides, alkyl sulfates, alpha olefin sulfonates, lignosulfonates, organophosphorous surfactants, and/or sarcosides. Nonionic surfactants include, but are not limited to, ethoxylated linear alcohols, ethoxylated alkyl phenols, ethoxylated thiols, acid ethoxylated fatty acids (polyethoxy-esters), glycerol esters, esters of hexitols and cyclic anhydrohexitols, ethoxylated amines, imidazoles, and/or tertiary amine oxides. Cationic surfactants include, but are not limited to, fatty amines, their salts and quaternary derivatives, linear diamines, amide, ester and ether amines, oxy and ethoxy amines, and/or alkanol amides. Buffers include, but are not limited to, phosphate, carbonate, acetate, and/or citrate buffers. In any embodiment, the potential at the working electrode may be used to help remove the molecule from the MIP film. For example, cycling between -IV to IV to extract the molecule from the polymer film. In any embodiment, if the molecule is a protein, the protein may be denatured and rinsed away from the polymer.

[0075] By combining molecules with polymers, a cavity remains in the polymer after removing the molecules. The cavities complement the molecule in size, shape, and chemical functionality. The cavities form the receptor sites for the molecules. Thus, the MIPs are solid or gel-phase polymers which were synthesized or deposited in the presence of a molecule. NIPs are synthesized with the same processes as MIPs but without the molecules.

[0076] The selective binding capabilities of the MIPs can be measured by incubating them in a solution of the tracer molecule and measuring how much binding occurs. In any embodiment, binding may be measured by cyclic voltammetry (CV), linear sweep voltammetry, square wave voltammetry, differential pulse voltammetry, amperometry, or a combination of two or more thereof. Binding behavior of the MIPs is compared with the NIPs. Methods of detecting binding in such systems include direct measurement of the film to observe the incorporation of bound tracer molecule. For lab-based challenge or standardization testing, the remaining tracer molecule in solution may also be used to indirectly measure binding. These measurements may be taken before and after incubation, continuously, or with some degree of mid-incubation data points. In any embodiment, changes between the pre-incubation and post-incubation measurements for the MIP and NIP control films indicate the presence or absence of the target molecule.

[0077] Electropolymerized chips may include additional components. For example, as disclosed herein the surface of the working electrode, the counter electrode, and/or the reference electrode may be modified.

[0078] The present technology provides a convenient method to detect molecule(s) such as allergens, pathogens, and/or toxins in a tangible good. In any embodiment, the present disclosure provides a method for detecting the molecule(s) using the detection device described herein, comprising exposing the sensor to the tangible good.

[0079] In any embodiment, the method of detecting the molecule, further includes: a) exposing the substrate to the tangible good; and b) inserting the substrate into the chamber. In any embodiment, upon inserting the substrate into the chamber, the substrate may puncture a capsule filled with solvent. Hence, in any embodiment the method comprising the steps of, the inserting the substrate into the chamber may puncture the capsule and release the liquid into the chamber. In any embodiment, the method may include agitating the device. In another embodiment, the agitating may include shaking, stirring, and/or grinding. In any embodiment, the agitating may continue until a substantially homogenous mixture is formed. [0080] After the tangible good sample has been inserted into the device, and the sample has been mixed with a solvent, the user may puncture a barrier separating the mixed sample and the sensor comprising the electropolymerized MIP film and electropolymerized NIP film. Then, the user inserts the exposed portion of the sensor (i.e., connector) into the processing device. Hence, in some embodiments, the method further comprises inserting the portion of the sensor outside of the body of the device into the processing device. Finally, the user can read the result of the processing device.

[0081] To enable convenient detection of the above listed molecules, the technology provided herein may be in the form of a wearable detection device.

[0082] The present technology, thus generally described, will be understood more readily by reference to the following examples, which are provided by way of illustration and are not intended to be limiting of the present invention.

EXAMPLES

[0083] The examples herein are provided to illustrate advantages of the present technology and to further assist a person of ordinary skill in the art with preparing or using the compositions of the present technology. The examples herein are also presented to more fully illustrate the preferred aspects of the present technology. The examples should in no way be construed as limiting the scope of the present technology, as defined by the appended claims. The examples can include or incorporate any of the variations, aspects, or aspects of the present technology described above.

[0084] Example 1 : Computational Complexation Energy. A pre-polymerization complex of the imprinted molecules with the one or more polymerized monomers disclosed herein was computationally optimized using the following method:

[0085] The structures of a given polymerized monomer and molecule were optimized separately using density functional theory (DFT) with a Becke, 3 -parameter, Lee- Yang-Parr (B3LYP) hybrid functional and a 6-31+G(d,p) Pople basis set. The total energy of the polymerized monomer-molecule complex was determined for different complex conformations by varying the internal rotational conformations of the monomer and molecule, the monomer-molecule intermolecular distance, and the intermolecular angles and dihedral angles using DFT with a B3LYP, PBE-type, or Minnesota exchange functional with a range of Pople basis sets from 3-21+G(d,p) to 6-311+G(d,p). More specifically, after selecting a basis set and level of theory, the electronic energies of both the monomer and the molecule were calculated separately. The sum of these two electronic energies was used as the baseline, representing a system with no complexation energy. Because the monomer and molecule may form complexes of different energies in different orientations, the following procedure was applied: The monomer was oriented at a fixed angle relative to a functional group on the molecule. An optimization was performed, keeping all molecular coordinates fixed except: a) the monomer-molecule distance; b) the bond angle and dihedral angle formed between the atoms of the monomer and a fixed atom on the monomer; c) the angles and dihedral angles between the atoms in the molecule functional group used to orient the monomer.

[0086] This process was repeated for a range of monomer-molecule angles from 60° to 120°, and with varying function groups of the molecule. The energies resulting from these calculations were tabulated, and used to probe for local energy minima. Starting near a local minima, an optimization was performed for which the monomer-molecule angle was allowed to vary, yielding a local minimum energy. The energy of the separate monomer and molecule (calculated at the same level of theory) (z.e., no-complexation energy) was subtracted from the local minimum energy to determine the energy of a single monomer- molecule interaction. The energies from all such interactions were added to yield the complexation energy (i.e., the change in electronic energy due to complexation). A more negative (greater magnitude) complexation energy corresponded to a stronger interaction between the monomer and molecule. A counterpoise correction was applied to account for the basis set superposition error.

[0087] To account for the energy of solvation and its effect on complexation, this process was repeated for a range of dielectric constants representing distinct solvents in the COSMO solvation model from 0 (gas-phase) to 78.4 (water). The complexation energy values were compared for various monomers disclosed herein and across different monomer to molecule ratios (1 : 1 to 4: 1). The monomers that produced more negative complexation energies typically yielded more selective/specific MIPs.

[0088] Example 2: 2-Aminophenol-based electropolymerized MIP and NIP sensor chips. Aminophenol and a peanut epitope were found to have a calculated complexation energy of -141 kJ/mol. Gold or carbon screen-printed electrodes (SPEs) were activated with 0.5 M H2SO4 (applying 5 - 15 cyclic voltammetric scans; potential range: -0.5 V to +1.5 V, scan rate: 100 mV s^-1 until a stable signal was achieved). Following activation, 10 pL of a peanut epitope solution (100 ppb -150 ppm in 1X-5X PBS; pH = 7.4) was placed on the working electrode of the SPE and left to incubate for 15 min - 24 h. The SPE was then gently rinsed with a 0.1 M phosphate buffer or phosphate-buffered saline (PBS) (pH 7.0- 8.0) and subsequently dried. A 100 pL of a 7-15 mM 2-aminophenol solution was placed on the SPE and electropolymerization was performed using cyclic voltammetry (1 - 10 scans; potential range: 0 V to +0.9 V, scan rate: 10-100 mV s^-1). The polymerization solution was then replaced with 100 pL of 0.1 M phosphate buffer (pH 7.4), and an additional 5 cyclic voltammetric scans, using the same parameters as before, were applied to remove unpolymerized monomer residue and to stabilize the polymer. The epitope was removed from the imprinted polymer by subjecting the MIP-coated electrodes to water for 24h. A control electrode (NIP) was prepared using the same procedure as for the MIP in the absence of the epitope.

[0089] For non-redox active molecules (e.g., epitopes and proteins), the target molecule rebinding was confirmed by measuring the degree of response (current intensity) of fabricated films in a buffer solution containing 1-10 ppm of the target molecule and a redox couple (5 mM; potassium ferricyanide/ferrocyanide, hexaammineruthenium (II)/(III) chloride, ferrocenecarboxylic acid (II)/(III), hydroquinone, ferrocene, or methylene blue, etc.) across the -0.6 V - 0.6 V potential range. For the redox marker to show redox activity, it approaches the electrode to facilitate charge transfer. The nonconductive polymeric matrix prevents the couple from approaching the electrode, inhibiting the redox reaction. In an imprinted film, unoccupied molecule binding sites allow charge transfer, whereas occupied binding sites do not. Therefore, the current response of the electrode to the redox probe is inversely related to the amount of molecule bound. Following this methods, the MIP successfully detected the presence of the peanut epitope after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope in IX PBS for five minutes.

[0090] Example 3: 2-Vinylpyridine-based electropolymerized MIP and NIP sensor chips. Gold or carbon screen-printed electrodes (SPEs) were activated, gently washed, and dried following the method of Example 2. Next, an epitope monomer solution was prepared by dissolving 1.0-5.0 mg of peanut epitope and 300-600 mg of 4-vinylpyridine (4- VP) in 3 mL of IX PBS solution (pH 7.0-8.0) containing 0.2 M potassium bromide (KBr). The SPEs were placed in the epitope monomer solution for 5-10 minutes followed by placing 100 pL of the epitope monomer solution on the SPE and preforming electropolymerization using cyclic voltammetry (5 - 15 scans; potential range: -1.0 V to 1.2 V, scan rate: 50 mV s^-1). The epitope was removed from the imprinted polymer by washing the MIP-coated electrode with a 0.1 M NaOH for two hours and left to dry for 24h at room temperature. A control electrode (NIP) was prepared using the same procedure as for the MIP in the absence of the epitope. Following the method in Example 2, the MIP successfully detected the presence of the peanut epitope after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope in IX PBS for five minutes.

[0091] Example 4: Acrylamide-based electropolymerized MIP and NIP sensor chips. Acrylamide and an epitope were found to have a calculated complexation energy of -104 kJ/mol. Gold or carbon screen-printed electrodes (SPEs) were activated, gently washed, and dried following the method of Example 1. Next, an epitope monomer solution was prepared by dissolving 5-10 mg of peanut epitope, 50-100 mg of N,N’- methylenebis(acrylamide), and 5-15 mg of acrylamide in 9 mL of IX PBS buffer (pH 7.4). Separately, a IX PBS solution containing 10-15 mg/mL ammonium persulfate (APS) and a IX PBS solution containing 20% Tetramethylethylenediamine (TEMED) were prepared. 1 mL of the APS solution was added into the epitope monomer solution followed by 5-15 minutes stirring. 10-50 pL of the TEMED solution was then added and the final solution was allowed to stir for an additional 1-5 minutes. The SPEs were placed in the final epitope monomer solution for 1-5 minutes followed by electropolymerization using cyclic voltammetry (10 - 20 scans; potential range: -0.4 V to 1.4 V, scan rate: 50-100 mV s^-1).

The epitope was removed from the imprinted polymer by washing the MIP-coated electrode with a 0.05-0.15 M NaOH for 1-5 hours, rinsed with distilled water, and stored in IX PBS solution. A control electrode (NIP) was prepared using the same procedure as for the MIP in the absence of the epitope. Following the method in Example 2, the MIP successfully detected the presence of the peanut epitope in a 1 ppm (1 mg/L) solution of the epitope in IX PBS for five minutes.

[0092] Example 5: Resorcinol -based electropolymerized MIP and NIP sensor chips. Resorcinol and an epitope were found to have a calculated complexation energy of -132 kJ/mol. Gold or carbon screen-printed electrodes (SPEs) were activated, gently washed, and dried following the method of Example 1. Following activation, 10 pL of a peanut or gluten epitope solution (100 ppb -150 ppm in 1X-5X PBS; pH = 7.4) was placed on the working electrode of the SPE and left to incubate for 15 min - 24 h. The SPE was then gently rinsed with a 0.1 M phosphate buffer or phosphate-buffered saline (PBS) (pH 7.0- 8.0) and subsequently dried. A 100 pL of a 5-15 mM resorcinol solution was placed on the SPE and electropolymerization was performed using cyclic voltammetry (1 - 15 scans; potential range: 0 V to +0.9 V, scan rate: 10-100 mV s^-1). The polymerization solution was then replaced with 100 pL of 0.1 M phosphate buffer (pH 7.4), and an additional 5 cyclic voltammetric scans, using the same parameters as before, were applied to remove unpolymerized monomer residue and to stabilize the polymer. The epitope was removed from the imprinted polymer by subjecting the MIP-coated electrodes to water for 24h. A control electrode (NIP) was prepared using the same procedure as for the MIP in the absence of the epitope. Following the method in Example 2, the MIP successfully detected the presence of the peanut and gluten epitopes after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitopes in IX PBS for five minutes.

[0093] Example 6: Hydroxy ethylmethacrylate-based MIP and NIP sensor chips.

Hydroxy ethylmethacrylate and an epitope were found to have a calculated complexation energy of -121 kJ/mol. Gold or carbon screen-printed electrodes (SPEs) were activated, gently washed, and dried following the method of Example 1. Next, a polymer solution was prepared by dissolving 5-10 mg of peanut epitope, 1-10 mg of geni stein, or 50-100 pg of lactose and 15-30 mg of poly(hydroxyethylmethacrylate) in 1 mL of ethanol. 1 pL of the polymer solution was drop-cast onto the SPE and left to dry at room temperature for 30 minutes. A control electrode (NIP) was prepared using the same procedure as for the MIP in the absence of the epitope. Following the method in Example 2, the MIP successfully detected the presence of the peanut epitope, genistein, and lactose after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope, genistein, or lactose in IX PBS for five minutes.

[0094] Example 7: Cross-linked hydroxy ethylmethacrylate-based electropolymerized MIP and NIP sensor chips. Gold or carbon screen-printed electrodes (SPEs) were activated, gently washed, and dried following the method of Example 1. Next, an monomer solution was prepared by mixing 15-25 mg hydroxy ethylmethacylate (HEMA) monomer, a 3 - 5 mg peanut epitope with 200-400 mg of a cross-linkers (polyethylene glycol) methyl ether methacrylate, ethylene glycol dimethacrylate (EGDMA), trimethyl tripropane triacrylate (TMPTA), or glycerol dimethacrylate) in 9 ml of a 0.1 M phosphate buffer, IX PBS, or water. 5-10 mg of peanut epitope, 1 ml of 13 mg/ml ammonium persulfate in IX PBS, and 50 pL of 5% Tetramethylethylenediamine (TEMED) in IX PBS (10 mL) were added to the monomer solution to provide an epitope monomer solution. The SPEs were placed in the epitope monomer solution for 5 minutes followed by electropolymerization using cyclic voltammetry (10 - 20 scans; potential range: -0.4 V to 1.4 V, scan rate: 50-100 mV s^-1).

The epitope was removed from the imprinted polymer by washing the MIP-coated electrode with a 0.05-0.20 M NaOH for 1-5 hours, rinsed with distilled water, and stored in IX PBS solution. A control electrode (NIP) was prepared using the same procedure as for the MIP in the absence of the epitope. Following the method in Example 2, the MIP successfully detected the presence of the peanut epitope after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope in IX PBS for five minutes.

[0095] Example 8: Over-oxidized polypyrrole-based electropolymerized MIP and NIP sensor chips. A monomer solution was prepared by adding 50-75 mg of pyrrole and 1-5 mg of milk epitope or casein into a beaker containing 10 ml of 0.1M KC1 solution. The resulting solution was stirred for 5 min. Polypyrrole based MIP films were grown electrochemically on the screen-printed electrodes (SPEs) using either cyclic voltammetry (potential range: 0 V to 1.0 V) or multistep amperometry (0 V for 0.5 - 2.0 sec followed by 0.6 V to 1.0 V for 0.5 - 2.0 sec) for 10 -100 cycles / pulses. The polypyrrole MIP coated electrodes were then over-oxidized in IX PBS solution by applying a potential pulse of 0.9 V for 180 sec. The electrodes were then washed in 1 :2 lOmM NaOH and ethanol solution for 3 hours to remove the epitope. The NIP (control) was prepared following the same protocol without any epitope added into the mixture. Following the method in Example 2, the MIP successfully detected the presence of the milk epitope and casein after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope or casein in IX PBS for five minutes.

[0096] Alternatively, over-oxidized polypyrrole-based electropolymerized MIP and NIP sensor chips were prepared by activating the SPEs following the procedure in Example 2, followed by incubating for 2 h in 50-200 ppm epitope or casein solution in IX PBS. These electrodes were then transferred to a beaker containing 10 ml of 0.2 M potassium phosphate dibasic, 50-75 mg of pyrrole, and 1-5 mg of epitope or casein. Electropolymerization was performed by applying a constant potential of 0.6 V - 0.9 V for 180-300 seconds. The polymer films were then washed in 10-25 mM NaOH for 3 hours to remove the epitope or casein. NIP films were prepared using the same methodology in the absence of epitope or casein. Following the method in Example 2, the MIP successfully detected the presence of the milk epitope and casein after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope or casein in IX PBS for five minutes.

[0097] Example 9: Dopamine-based electropolymerized MIP and NIP sensor chips. Prior to electropolymerization, the SPEs were pretreated using the procedure described above (applying 5 - 15 cyclic voltammetric scans; potential range: -0.5 V to +1.5 V, scan rate: 100 mV s-1 in 0.5 M H2SO4 until a stable signal was achieved). The resulting electrodes were then incubated for 1-6 h in 1-200 ppm peanut or gluten epitope solution in IX PBS or 0.1 M phosphate buffer, and washed with water and IX PBS. The monomer solution, containing 5 mM dopamine dissolved in IX PBS (pH = 7.4), was degassed for one hour prior to the polymerization.

[0098] Electropolymerization was performed by drop-casting 100 pl of the monomer solution on the pretreated electrodes, and cycling the potential between -0.5 V to +0.5 V for 5 to 30 cycles at 10 -100 mV/sec. The resulting polymer films were washed with water and then rinsed with PBS to remove any unreacted monomer from the surface. The epitope was extracted from the imprinted polymer film by cycling the potential from -0.1 V to 0.9 V for 1-10 cycles at 100 mV/sec in IX PBS. NIP films were prepared using the same methodology in the absence of epitope. Following the method in Example 2, the MIP successfully detected the presence of the epitope after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope in IX PBS for five minutes.

[0099] Example 10: Aminobenzoic acid-based electropolymerized MIP and NIP sensor chips. Prior to the electropolymerization, the SPEs were pretreated using the procedure described above (applying 5 - 15 cyclic voltammetric scans; potential range: -0.5 V to +1.5 V, scan rate: 100 mV s-1 in 0.5 M H2SO4 until a stable signal was achieved). Cleaned electrodes used for MIP polymerization were subsequently incubated for 1-6 h in 1-200 ppm epitope solution in IX PBS or 0.1 M phosphate buffer, and then washed with water and IX PBS. Electropolymerization was performed by drop-casting 100 pl of the monomer solution (5 mM aminobenzoic acid in 0. IM PB (pH = 7.4)) on the pretreated electrodes, and cycling the potential between -0.5 V to +1.5 V for 5 to 20 cycles at 10 -100 mV/sec. The resulting polymer films were washed with water and then rinsed with IX PBS (pH = 7.4) to remove any unreacted monomer from the surface. The epitope was extracted from the imprinted polymer film by cycling the potential from -0.6 V to 1.4 V for 1-10 cycles at 100 mV/sec in IX PBS or 0. IM PB. NIP films were prepared using the same methodology in the absence of epitope. Following the method in Example 2, the MIP successfully detected the presence of the epitope after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope in IX PBS for five minutes.

[0100] Example 11 : Scopol etin-based electropolymerized MIP and NIP sensor chips. Before modification, the gold electrode was subjected to potential cycling in 0.5 M H2SO4 for 10 cycles (potential range: -0.5 V- 1.5 V; scan rate: 100 mV/sec). The resulting electrodes were rinsed with deionized water and PBS solution (IX, pH = 7.4), and then incubated in 1 ppm of epitope solution for one hour. The epitope modified electrode was dipped into the 0.1 M PB or IX PBS solution (pH = 7.4) containing 0.1 - 5 mM scopoletin as monomer. Electropolymerization was used to form molecular imprinted film by cyclic voltammetry from -0.2 V to 0.7 V for 10 cycles to obtain rigid, uniform, and compact MIP film. The template was eluted by subjecting the polymer film to either an alkaline (0.1 M NaOH), or acidic wash (0.1 M H2SO4). Following the method in Example 2, the MIP successfully detected the presence of the epitope after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope in IX PBS for five minutes.

[0101] Example 12: PEDOT-based electropolymerized MIP and NIP sensor chips. The imprinted polymer was synthesized on the previously pretreated electrodes (as described above), by electropolymerizing a 5 mM solution of 3, 4-ethylenedi oxythiophene (EDOT) prepared in 0.1 M PB or IX PBS buffer on the epitope modified electrode. Polymerization was carried out by CV, between -0.2 V and +0.90 V, at 100 mV/sec, for 5-10 cycles. Following the method in Example 2, the MIP successfully detected the presence of the epitope after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope in IX PBS for five minutes.

[0102] Example 13: O-phenylenediamine-based electropolymerized MIP and NIP sensor chips. A pretreated gold electrode was incubated for 30-60 minutes in a phosphate buffer solution containing 40-100 mg of o-phenylenediamine and 1-10 mg of peanut epitope, genistein, daidzein, lactose, quercetin, tryptophan, or biochanin A. Afterwards, electropolymerization was performed on the electrode through cyclic voltammetry for 5-20 cycles by varying applied potential from 0 V to 1.0 V at scan rate of 100 mV/sec. The electrode was then washed with 0.1 M NaOH for 30-120 min to remove the molecule. It was then rinsed with distilled water and stored in IX PBSuntil further use. Following the method in Example 2 for the epitope, the MIP successfully detected the presence of the epitope after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope in IX PBS for five minutes.

[0103] Following the method in Example 2 for the epitope, the MIP successfully detected the presence of the epitope after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope in IX PBS for five minutes. For genistein, daidzein, lactose, quercetin, tryptophan, and biochanin A (i.e., redox-active molecules), the faradaic current was measured based on the direct electron transfer between genistein, daidzein, lactose, quercetin, tryptophan, or biochanin A and the underlying electrode. [0104] Example 14: Copolymer-based electropolymerized MIP and NIP sensor chips Imprinted films consisting of two or more different polymer subunits were prepared by dissolving each monomer (combination of monomers listed above) at optimized concentration in the same flask. Prior to electropolymerization, the electrodes were then incubated for 1-6 h in 1-200 ppm milk, peanut, or gluten epitope solution in IX PBS or 0.1 M phosphate buffer, and washed with water and IX PBS. Electropolymerization was conducted following any of the above methods. The milk, peanut, or gluten epitope was removed from the copolymer film by either washing the MIP-coated electrodes in acidic, neutral, or alkaline conditions for 1-5 hours under rigorous stirring (700-1200 rpm), or by cycling the potential in the 0.1 M phosphate buffer or IX PBS buffer (pH = 7.4) from -0.2 V - 0.9 V for 1-10 cycles. Following the method in Example 2, the MIP successfully detected the presence of the milk, peanut, and gluten epitope after incubation of the electrode in a 1 ppm (1 mg/L) solution of the epitope in IX PBS for five minutes.

[0105] Example 15. Preparation of epitope-imprinted polymer sensors. All sensors were prepared using either gold or carbon screen-printed electrodes (SPEs). In some instances, carbon-based SPEs were used that had been modified with carboxyl or amine functionality on the surface. For these electrodes, the electrodes were used as-is.

[0106] Step 1. A. Electrode grafting on carbon SPEs (amine-grafting). A total of 400 pL of 0.1 sodium nitrite solution in 0.5 M HC1 (final concentration: 2 mM) was added to a 20 mL solution containing 2 mM phenylenediamine (4-aminobenzylamine, 2- aminobenzylamine, 4-(2-aminoethyl)benzylamine, N-methyl-l,2-benzylamine, or N,N- dimethyl-p-benzylamine) and 0.5 M HC1 under stirring. The mixture was stirred for 5-10 min prior to electrochemical grafting.

[0107] Electrochemical reduction of in situ generated aminophenyl monodiazonium cations was performed by placing 100 pL of the solution onto the electrode and applying constant potential (-0.3 V to -1.0 V range) for 5 to 360 seconds. The resulting electrodes were rinsed with ultra-pure water for at least 5 seconds and then stored in either 0.1 M PB, 0.1 M PBS buffer solution, or ultra-pure water until further use. [0108] B. Electrode grafting on carbon SPEs (carboxyl-grafting). A solution of 5 mL of 4 mM 4-aminobenzoic acid or 4-amino-2-methylbenzoic acid in 0.5 M HC1 was added directly into 5 mL of sodium nitrite solution (4 mM) in 0.5 M HC1 and stirred for 10 min (final concentration - 2 mM). Electrografting was performed via cyclic voltammetry by sweeping the potential from +0.2 V to -0.6 V at 10 - 100 mV/sec for 1-3 cycles. The fabricated electrodes were rinsed with ultra-pure water for at least 5 seconds to remove any unbound reactants and then stored in either 0.1 M PB, 0.1 M PBS buffer solution or ultra- pure water until further use.

[0109] Gold electrode functionalization with amines or carboxyl groups. SPEs were pretreated by applying 5-15 cyclic voltammetric scans over a potential range of -0.5 V to +1.5 V, at a scan rate: 100 mV s^-1 in 0.5 M H2SO4, until a stable signal was achieved. The resulting electrodes were then incubated for 1-24 hours in cysteamine, 3 -mercaptopropionic acid (100 ppb - 1000 ppm of the reagent in IX PBS, 0.1 M phosphate buffer or water), or aminothiophenol (100 ppb - 1000 ppm in ethanol), and washed with ultrapure water, buffer, or ethanol. The electrodes were then stored in water prior to surface functional group activation.

[0110] Step 2. Activation of electrografted surfaces and epitope incubation. Glutaraldehyde step and epitope incubation for aminobezyl amine functionalized electrodes. 100 pL of 50% glutaraldehyde (GA) solution (in water) was added to 900 pL of 0.1 M PB or PBS buffer (pH is from 6-9) and stirred together in the dark for 1 min. Then, 30 pL of the resulting 5% GA solution was drop-cast onto the working electrode and left on a shaker (under gentle shaking) for 0.5-3 hours in the dark. Subsequently, the resulting electrodes were rinsed with the same buffer as used for GA incubation. A 10 pL sample of a cleavable epitope solution (10-1000 ppm of the epitope in 0.1 M PB or 0.1 M PBS buffer at pH ranging from 7.0 - 8.5) was drop-cast directly onto the electrodes and left to incubate for 1- 3 hours. After incubation, the epitope-modified electrodes were rinsed with ultra-pure water to remove any physiosorbed epitope from the surface. In some instances, the unreacted GA groups were deactivated / capped by immersing the electrodes in either a 0.1 M ethanolamine solution (in 0.1 M PB or PBS buffer; pH = 7 - 8.5) or 0.1 M Tris buffer solution (pH = 8.5) for 1 hour. Each electrode was then rinsed thoroughly with ultra-pure water prior to electropolymerization.

[0111] EDC/NHS activation step and epitope incubation for aminobezoic acid functionalized electrodes. The carboxyl groups on aminobenzoic acid-modified carbon electrodes were then activated through drop-casting 100 pL of A-(3 -di methyl ami nopropyl )- TV'-ethylcarbodiimide hydrochloride (EDC) (50 - 400 mM) and A-hydroxysuccinimide (NHS) or A-hydroxysulfosuccinimide sodium salt (NHS salt) (10-400 mM) in 0.1M MES buffer at pH 5.5. After 1 h, the activated electrodes were rinsed with 0.1 M MES buffer (pH = 5.5) or ultra-pure water and dried under air. Subsequently, 10 L of a cleavable epitope (10-1000 ppm of the epitope in 0.1 M PB or 0.1 M PBS buffer at pH 7.0-8.5) solution was drop-cast directly onto the electrodes. The electrodes were left to incubate for 1-3 hours. After incubation, the epitope-modified electrodes were rinsed with ultra-pure water to remove any physiosorbed epitope from the surface. In some instances, the unreacted EDC/NHS groups were deactivated / capped by immersing the electrodes in either a 0.1 M ethanolamine solution (in 0.1 M PB or PBS buffer; pH = 7 - 8.5) or 0.1 M Tris buffer solution (pH = 8.5) for 1 h. Following this step, each electrode was rinsed thoroughly with ultra-pure water prior to the electropolymerization.

[0112] Step 3. Imprinted polymer formation. Electropolymerization of imprinted polymers was performed. Non-imprinted polymers are prepared using the same methodology in the absence of the epitope or with the “non-cleavable” version of the same epitope. For example, if the imprinted polymer was prepared using a generically described KC-CRRRRRRRRRR epitope (- refers to a cleavable intermolecular disulfide bond between two different peptides) then for the NIP preparation a KCCMRRRRRRR polypeptide having a peptide bond may be used instead of the disulfide bridge.

[0113] Example 16. Dopamine synthesis. The monomer solution, containing 5 mM dopamine dissolved in IX PBS or 0.1 M PB (pH = 7.4) buffer solution, was degassed with nitrogen for one hour prior to the polymerization. Electropolymerization was then performed by drop-casting 100 pl of the monomer solution on the epitope-functionalized electrodes, and cycling the potential between -0.65 V to +0.65 V for 5 to 30 cycles at 10 - 100 mV/sec. The resulting polymer films were washed with ultra-pure water and then rinsed with 0.1 PB or IX PBS (pH = 7.4) to remove any unreacted monomer from the surface.

[0114] Example 17. m-Phenylenediamine (mPD) synthesis. Electropolymerization was performed by drop-casting 100 pl of the monomer solution (5 - 10 mM mPD in 0. IM PB or PBS (pH = 7.4) on the electrodes, and applying constant potential of 0.6 V for 1 - 60 seconds. The resulting polymer films were washed with water and then rinsed with IX PBS or 0.1 M PB (pH = 7.4) to remove any unreacted monomer from the surface.

[0115] Norepinephrine or dopamine synthesis (air oxidation): a 1-5 mg/ml solution of norepinephrine or dopamine in either 0.1 M PB, PBS buffers (pH = 7 - 10) or 0.1M Tris buffer (pH = 8.5) was drop-casted onto the epitope-functionalized working electrodes immediately after the preparation. The electrodes were kept in dark under humid environment (to avoid liquid evaporation from the electrode surface) for 1-36 hours. After polymer synthesis, the films were rinsed with ultra-pure water.

[0116] The epitope was extracted from the imprinted polymer film by breaking the intermolecular disulfide bond connecting the two peptide molecules with a reducing agent. Specifically, the polymer-film coated electrodes were placed in a 1-50 mM solution of dithiothreitol (DTT), 2-mercaptoethanol, tris (2-carboxyethyl) phosphine hydrochloride (TCEP), sodium borohydride, thioglycolic acid, or di thioerythritol dissolved in 0.1 M PB or IX PBS (pH range from 7.0 to 8.5). Because of higher stability of TCEP to degradation under different pH, additional reducing solutions may include 1-50 mM TCEP in ultra-pure water, 0.1 M HC1, acetate buffer, carbonate buffer, HEPES, MES and Tris buffers. Alternatively, disulfide bonds reductions can be carried out by placing zinc dust / nanoparticles in 1-5% of acetic acid solution. The reduction time ranges from 1 min-24 hours. In some instances, the template extraction is performed at elevated temperature (20°C to 80°C under stirring). Following disulfide bond reduction, the electrodes can be subjected to additional wash to remove the residual epitope from the film. These washes may include: buffer solution (e.g., PB, PBS, Tris) with an added cationic, zwitterionic or anionic surfactant (0.1 to 5% by weight); treatment with basic (sodium hydroxide) or acidic solutions (acetic, formic, oxalic, hydrochloric, sulfuric acids).

[0117] Example 18. Probe Detection. The same detection protocol and principles as above may be applied. For the measurement of electroinactive analytes by MIP based sensors, redox-active labels / “tracers” can be used to generate an electrochemical signal - a redox couple with a well-established oxidation / reduction potential. The removal of the template molecule (e.g., epitope, protein) generates pathways in the tight MIP layer, which allow the permeation of the redox marker to the electrode surface to provide a current signal via its oxidation or reduction. Rebinding of the target will decrease the current signal by closing these pores / cavities and subsequently the pathways to the electrode, therefore causing a concentration-dependent decrease in the permeation of the redox marker. This methodology typically applies for insulating MIPs that constrain the redox reaction of the marker probe at the electrode surface.

[0118] Example 19. A MIP-coated electrode was placed in 0.1 M PBS (pH 7.0 - 8.0) solution containing different concentrations of the epitope (10 ppb - 100 ppm) and 5 mM of the redox probe, which was either potassium ferricyanide/ferrocyanide, hexaammineruthenium (II)/(III) chloride, ferrocenecarboxylic acid (II)/(III), hydroquinone, ferrocene, or methylene blue. Electrochemical measurements were performed with either cyclic voltammetry (1-10 scans; scan rate: 10 mV s^-1 - 1 V s^-1) or differential pulse voltammetry (DPV).

[0119] Example 20. Post-synthetic modifications. An issue in producing biosensors is the prevention of non-specific binding for accurate detection. Non-specific binding occurs when an interfering molecule adsorbs to the surface of a sensor, resulting in high background signal or sensor fouling. To reduce non-specific binding, physical methods, chemical methods, or both may be used. Physical methods of antifouling examined include the use of blocking agents such as bovine serum albumin, casein, gelatin, or ovalbumin (0.1 - 5% by weight in 0.1 M or IX PBS solution). Surface blocking against non-specific binding is carried out by drop-casting of a blocking solution directly on a surface of MIP - coated electrodes for 5 min to 3 hours. The electrodes are then thoroughly rinsed with the buffer solution or ultra-pure water prior to further use.

[0120] In addition, chemical blocking methods may be based on direct functionalization with poly(dopamine) and poly(norepinephrine) through grafting MIPs surface with aminoterminated or thiol -terminated molecules via Michael addition and Schiff base reactions. For example, fouling-resistant surfaces are prepared by covalently attaching thiol- terminated methoxy -poly(ethylene glycol) or ethanolamine either before or after template extraction. To achieve thiol -terminated methoxy-poly(ethylene glycol) functionalization of poly(dopamine) film, MIP or NIP-coated electrodes are placed in a 10 - 100 mM solution of the reactant in 0.1 M PB or IX PBS (pH = 7-9) for 1-3 hour under stirring (T = 20-80 °C). Following incubation, the resulting electrodes are rinsed with ultra-pure water or buffer solution prior to the electrochemical characterization.

[0121] Epitope selection. To form a cleavable disulfide bond between two peptide molecules, each peptide must contain a thiol -based moiety at either N- or C-terminus. For this purpose, epitopes that contain a terminal cysteine may be used, or a cysteine amino acid may be added to the peptide during synthesis. After coupling, the resulting polypeptide consisted of the targeted epitope and a “linker” peptide. The “linker” peptide is covalently attached to the electrode surface via EDC/NHS or glutaraldehyde chemistries. The linker molecule can be a peptide that contains suitable functional groups for surface attachment using methods described above (primary amines, lysine amino acids, etc.) together with a thiol-based molecule needed to form an intermolecular disulfide bond. The simplest molecule of this kind can be a KC (lysine-cysteine) peptide. However, the length of the peptide can be further extended by incorporating more amino acids between the terminal amino acids, if required. Alternatively, the linker peptide can be replaced with cysteamine.

[0122] Other chemistries (e.g., click chemistry) can be used for coupling epitopes with amine- or carboxyl-functionalized electrodes.

[0123] While certain embodiments have been illustrated and described, a person with ordinary skill in the art, after reading the foregoing specification, can effect changes, substitutions of equivalents, and other types of alterations to the compositions of the present technology as set forth herein. Each aspect and embodiment described above can also have included or incorporated therewith such variations or aspects as disclosed in regard to any or all of the other aspects and embodiments.

[0124] The present technology is also not to be limited in terms of the particular aspects described herein, which are intended as single illustrations of individual aspects of the present technology. Many modifications and variations of this present technology can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods within the scope of the present technology, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. It is to be understood that this present technology is not limited to particular methods, reagents, compounds, or compositions, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to be limiting. Thus, it is intended that the specification be considered as exemplary only with the breadth, scope, and spirit of the present technology indicated only by the appended claims, definitions therein and any equivalents thereof.

[0125] The embodiments, illustratively described herein, may suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. Thus, for example, the terms “comprising,” “including,” “containing,” etc. shall be read expansively and without limitation. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the claimed technology. Additionally, the phrase “consisting essentially of’ will be understood to include those elements specifically recited and those additional elements that do not materially affect the basic and novel characteristics of the claimed technology. The phrase “consisting of’ excludes any element not specified. [0126] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the invention. This includes the generic description of the invention with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein.

[0127] As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a nonlimiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like, include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member.

[0128] All publications, patent applications, issued patents, and other documents (for examplejournals, articles and/or textbooks) referred to in this specification are herein incorporated by reference as if each individual publication, patent application, issued patent, or other document was specifically and individually indicated to be incorporated by reference in its entirety. Definitions that are contained in text incorporated by reference are excluded to the extent that they contradict definitions in this disclosure.

[0129] Other embodiments are set forth in the following claims, along with the full scope of equivalents to which such claims are entitled.

Claims

WHAT IS CLAIMED IS:

1. A detection device comprising a sensor, wherein the sensor comprises: a circuit board; an electropolymerized molecularly imprinted polymer (MIP) film comprising one or more receptor sites imprinted in the polymer, the one or more receptor sites configured to accept an epitope, molecule, part of a molecule, and/or a combination of molecules; and an electropolymerized non-imprinted polymer (NIP) film; wherein: the sensor is configured to detect the presence of the molecule upon binding to one or more of the receptor sites; and the electropolymerized MIP film and the electropolymerized NIP film comprise one or more polymerized monomers selected from the group consisting of 3,4- ethylenedi oxy thiophene or a monomer of formula I, II, III, IV, V, VI, VII, VIII, or IX:

53

wherein:

A, B, D, E, and G are independently CH or N, provided that at least one of A, B, D, E, and G are N;

J is N or S;

R¹, R², and R³ are independently H or NH2;

R⁴, R⁵, and R⁷ are independently H or CH3;

R⁸ is OH or NH2;

R⁹ is absent, H, or CH3;

R¹⁰ and R¹¹ are independently H or C1-C5 alkylene-NH2;

54 R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ are independently H, NH2, or OH, provided that: at least one of R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ is NH2 or OH, or one of R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ is NH2 or OH;

R¹⁵ C1-C5 alkyl;

R¹⁶, R¹⁷, and R¹⁸ are independently H, NH2, or SH; and m is 1, 2, 3, 4 or 5. device of claim 1, wherein the molecule comprises an allergen, a trace molecule of an allergen, a pathogen, a toxin, or a combination of any two or more thereof. device of claim 2, wherein the allergen and/or the trace molecule of the allergen comprises an organic molecule or a salt thereof. device of claim 3, wherein the organic molecule is lactose, galactose, amygdalin, juglone, biochanin A, resveratrol daidzein, daidzin, genistein, or genistin. device of claim 3, wherein the organic molecule comprises a polypeptide, protein, epitope, aptamer, or a combination of any two or more thereof. device of claim 5, wherein the organic molecule comprises at least one protein. device of claim 5 or claim 6, wherein the organic molecule comprises at least two different proteins. device of claim 5, wherein the organic molecule comprises at least one epitope. device of claim 8, wherein the organic molecule comprises at least two different epitopes. e device of any one of claims 5-9, wherein the organic molecule comprises at least one protein and at least one epitope. e device of any one of claims 5-10, wherein the epitope comprises an Ara Hl epitope, Ara H2 epitope, Ara H3/H4 epitope, Ara H6 epitope, or a combination of any two or more thereof.

55 device of any one of claims 3-11, wherein the organic molecule is not cortisol, an amino acid, theophylline, and/or chlorpyrifos. device of any one of claims 2-12, wherein the pathogen comprises a food pathogen, a clinical pathogen, or a combination of any two or more thereof. device of claim 13, wherein the food pathogen comprises Campylobacter,

Cyclospora, Clostridium botulinum, Escherichia coli, Listeria, Salmonella, Staphylococcus aureus, Shigella, Toxoplasma gondii, Vibrio vulnificus, Norovirus, Hepatitis A, or a combination of any two or more thereof. device of claim 13 or claim 14, wherein the clinical pathogen comprises Candida,

Chlamydia trachomatis, Neisseria gonorrhoeae, Methicillin-resistant Staphylococcus aureus, Mycobacterium tuberculosis, human papillomavirus (HPV), Hepatitis B, Hepatitis C, Hepatitis D, Hepatitis E, human immunodeficiency virus, influenza, or a combination of any two or more thereof. device of any one of claims 1-15, wherein the toxin comprises a herbicide, a pesticide, a drug of abuse, or a combination of any two or more thereof. device of claim 16, wherein the herbicide or the pesticide comprises atrazine, azinphos-methyl, bentazone, carbaryl, carbofuran, chlorpyrifos methyl, chlorsulfuron, cyhexatin, diazinon, dimethoate, fenobucarb, glyphosate, hydrazine, imidacloprid, lindane, methyl parathion, paraquat, parathion, permethrin, pirimicard, sulfentrazone, or a combination of any two or more thereof. device of claim 16 or claim 17, wherein the drug of abuse comprises an amphetamine, cocaine, a benzodiazepine, a barbiturate, a dissociative drug, an opioid, a salt thereof, a metabolite thereof, or a combination of any two or more thereof. device of any one of claims 1-18, wherein the one or more polymerized monomers comprise a monomer of formula I.

56 device of claim 19, wherein R¹ is NH2, R² is H, and R³ is H. device of claim 19, wherein R¹, R², and R³ are H. device of any one of claims 1-21, wherein the one or more polymerized monomers comprise a monomer of formula II. device of claim 22, wherein R⁴ is H; A, B, E, and G are CH; and D is N. device of any one of claims 1-23, wherein the one or more polymerized monomers comprise a first monomer and a second monomer of formula III. device of claim 24, wherein R⁵ is H and R⁶ is H in the first monomer and R⁵ is H

the second monomer. device of any one of claims 1-25, wherein the one or more polymerized monomers comprise a monomer of formula IV. device of claim 26, wherein R⁷ is CH3, R⁸ is OH, and m is 2. device of any one of claims 1-27, wherein the one or more polymerized monomers comprise a monomer of formula V. device of claim 28, wherein J is N and R⁹ is H. device of claim 28, wherein J is S and R⁹ is absent. device of any one of claims 1-30, wherein the one or more polymerized monomers comprise a monomer of formula VI. device of claim 31, wherein R¹⁰ is H and R¹¹ is H or CH2-CH2-NH2. device of any one of claims 1-32, wherein the one or more polymerized monomers comprise a monomer of formula VII. device of claim 33, wherein R¹², R¹³, R¹⁹, and R²⁰ are H and R¹⁴ is NH2. device of claim 33, wherein R¹² and R²⁰ are H and R¹³, R¹⁴, and R¹⁹ are OH. device of any one of claims 1-35, wherein the one or more polymerized monomers comprise a monomer of formula VIII. device of claim 36, wherein R¹⁵ is CH3. device of any one of claims 1-37, wherein the one or more polymerized monomers comprise 3, 4-ethylenedi oxythiophene. device of any one of claims 1-38, wherein the one or more polymerized monomers comprise a monomer of formula IX. device of claim 39, wherein R¹⁶ is NH2 and R¹⁷ and R¹⁸ are each H. device of claim 39, wherein R¹⁶, R¹⁷, and R¹⁸ are each H. device of claim 39, wherein R¹⁶ is SH, R¹⁷ is H, and R¹⁸ is H. device of any one of claims 1-42, wherein the one or more polymerized monomers further comprises a cross-linker. device of claim 43, wherein the cross-linker comprises ethylene glycol dimethacrylate (EGDMA), trimethyl tripropane triacrylate (TMPTA), glycerol dimethacrylate, or a combination of any two or more thereof. device of any one of claims 1-44, wherein the electropolymerized MIP and the molecule have a complexation energy of less than 0 kJ/mol per binding site. device of any one of claims 1-45, wherein the electropolymerized MIP and the molecule have a complexation energy about -10 kJ/mol to about -150 kJ/mol per binding site. device of any one of claims 1-46, wherein the molecule is in salt form; and the one or more polymerized monomers comprise a monomer of formula V doped with F", Br", Cl", NCh", C1O₄", SO₄ ²’, PO4^3-, or a combination of any two or more thereof. device of any one of claims 1-47, wherein the molecule has a molecular weight less than about 1000 g/mol. device of any one of claims 1-48, wherein the molecule is hydrophobic and the one or more polymerized monomers comprise a monomer of formula I, II, V, VI, VII, or IX. device of any one of claims 1-49, wherein the molecule has one or more aromatic groups and the one or more polymerized monomers comprise 3,4- ethylenedi oxy thiophene or a compound of formula I, II, V, VI, VII, VIII, or IX. device of any one of claims 1-50, wherein the electropolymerized MIP and/or the electropolymerized NIP further comprise a tracer. device of claim 51, wherein the tracer is selected from the group consisting of potassium ferricyanide/ferrocyanide, hexaammineruthenium (II)/(III) chloride, ferrocenecarboxylic acid (II)/(III), hydroquinone, ferrocene, methylene blue, iridium (II)/(III) chloride, ascorbic acid, dopamine, and a combination of any two or more thereof. device of claim 51 or claim 52, wherein the electropolymerized MIP and/or the electropolymerized NIP has about 1 nmol to about 500 mmol of the tracer. device of any one of claims 51-53, wherein the electropolymerized MIP and/or the electropolymerized NIP has about 1 pmol to about 500 mmol of the tracer. device of any one of claims 1-54 further comprising an electrochemical chip, wherein the electrochemical chip comprises the electropolymerized MIP and the electropolymerized NIP.

59 device of any one of claims 1-55 further comprising a first electrochemical chip and a second electrochemical chip, wherein the first electrochemical chip comprises the electropolymerized MIP and the second electrochemical chip comprises the electropolymerized NIP. device of claim 55 or 56, the circuit board comprises the electrochemical chip or the first electrochemical chip and the second electrochemical chip. device of any one of claims 1-57 further comprising a processing device, wherein the processing device is configured to communicatively couple to the sensor, wherein the processing device is configured to determine an electric current difference and/or electric potential difference between the electropolymerized MIP film and the electropolymerized NIP film. device of claim 58, wherein the processing device determines the presence of the molecule when the electric current and/or potential difference of the electropolymerized MIP film is greater than the electric current and/or potential difference of the electropolymerized NIP film. device of claim 58 or claim 59, wherein the electric current and/or potential difference is determined by cyclic voltammetry (CV), linear sweep voltammetry, square wave voltammetry, differential pulse voltammetry, amperometry, or a combination of any two or more thereof. device of any one of claims 1-60 further comprising a chamber, wherein the chamber comprises a capsule that encapsulates a solvent and the chamber provides a space for mixing the solvent with a tangible good. device of claim 61, wherein solvent is water, a buffer, a saline solution, an organic solvent, or a combination of any two or more thereof. device of claim 64 or claim 65 further comprising a barrier separating the chamber and the sensor.

60 device of claim 63, wherein puncturing the barrier exposes the chamber comprising the solvent and tangible good to the sensor. device of any one of claims 1-64, wherein the sensor is configured to for insertion into a processing device. device of any one of claims 1-65, wherein the electropolymerized MIP film has a thickness of about 2 nm to about 100 nm. device of any one of claims 1-66, wherein the electropolymerized MIP film has a thickness of about 2 nm to about 20 nm. device of any one of claims 1-67, wherein the molecule is in a tangible good. device of claim 68, wherein the tangible good comprises food, drink, cosmetic, or a combination of any two or more thereof. ethod of making the detection device of any one of claims 1-69, the method comprising: providing a conductive electrode; depositing a polymer comprising the one or more polymerized monomers in the presence of the molecule by electropolymerization to form the electropolymerized MIP film; and depositing the polymer in the absence of the molecule by electropolymerization to form the electropolymerized NIP film. method of claim 70, wherein the depositing the polymer in the presence of the molecule is on an electrochemical chip. method of claim 71, wherein the depositing the polymer in the absence of the molecule is on the same or different electrochemical chip. method of any one of claims 71 or 72, wherein the molecule is immobilized to the electrochemical chip surface prior to depositing the polymer comprising the one or

61 more polymerized monomers in the presence of the molecule by electropolymerization to form the electropolymerized MIP film. detection device of any one of claims 1-69 or the method of any one of claims 70- 73, wherein the one or more polymerized monomers are determined by computationally calculating complexation energy of the one or more polymerized monomers and the molecule. detection device or the method of claim 74, wherein the computationally calculated complexation energy is less than zero. etection device comprising a sensor, wherein the sensor comprises: a circuit board; an electropolymerized molecularly imprinted polymer (MIP) film comprising one or more receptor sites imprinted in the polymer, the one or more receptor sites configured to accept a target molecule; and an electropolymerized non-imprinted polymer (NIP) film; wherein: the sensor is configured to detect the presence of the target molecule in a sample, when the sensor is exposed to the sample, upon binding to one or more of the receptor sites; and the electropolymerized MIP film and the electropolymerized NIP film comprise one or more polymerized monomers selected from the group consisting of 3,4- ethylenedi oxy thiophene or a monomer of formula I, II, III, IV, V, VI, VII, VIII, or IX:

wherein:

J is N or S;

R¹, R², and R³ are independently H or NH2;

R⁴, R⁵, and R⁷ are independently H or CH3;

R⁸ is OH or NH2;

R⁹ is absent, H, or CH3;

R¹⁰ and R¹¹ are independently H or C1-C5 alkylene-NH2;

63 R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ are independently H, NH2, or OH, provided that: at least one of R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ is NH2 or OH, or one of R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ is NH2 or OH;

R¹⁵ C1-C5 alkyl;

R¹⁶, R¹⁷, and R¹⁸ are independently H, NH2, or SH; and m is 1, 2, 3, 4 or 5. device of claim 76, wherein the electropolymerized MIP film has a thickness of about 1 nm to about 100 nm. device of claim 77, wherein the electropolymerized MIP film has a thickness of about 2 nm to about 20 nm. device of any one of claims 76-78, wherein the target molecule is an epitope. rocess for preparing a detection device, the process comprising: depositing on a sensor a template molecule on a portion of a surface of the sensor such that the template molecule forms a self-assembled monolayer on the sensor in a substantially perpendicular orientation to the surface to form a modified sensor; electropolymerizing a polymer film on the modified sensor forming both imprinted regions where the template molecule is present and non-imprinted regions where the template molecule is absent; and removing the template molecule from the sensor, such that a cavity is formed where the template molecule was present, the cavity being a specific binding pocket for a target molecule of the same as or similar to the template molecule; wherein the polymer film is the electropolymerization product of one or more of 3,4- ethylenedioxythiophene, or a monomer of formula I, II, III, IV, V, VI, VII, VIII, or IX:

64

wherein:

J is N or S;

R¹, R², and R³ are independently H or NH2;

R⁴, R⁵, and R⁷ are independently H or CH3;

65

R⁸ is OH or NH2;

R⁹ is absent, H, or CH3;

R¹⁰ and R¹¹ are independently H or C1-C5 alkylene-NH2;

R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ are independently H, NH2, or OH, provided that: at least one of R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ is NH2 or OH, or one of R¹², R¹³, R¹⁴, R¹⁹, and R²⁰ is NH2 or OH;

R¹⁵ C1-C5 alkyl;

R¹⁶, R¹⁷, and R¹⁸ are independently H, NH2, or SH; and m is 1, 2, 3, 4 or 5. method of claim 80, wherein the polymer film has a thickness of about 1 nm to about 100 nm. method of claim 81, wherein the polymer film has a thickness of about 2 nm to about 20 nm. method of any one of claims 80-82, wherein the removing comprises swelling the film. method of any one of claims 80-83, wherein the removing comprises cleaving a bond between a terminal end of the template molecule and the surface, such that the cavity is of sufficient dimensions to be specific to the target molecule. method of any one of claims 80-84, wherein the template molecule comprises an epitope.

66