Implantable device, biosensor device, system and method

Abstract

The present invention relates to biosensor devices, implantable devices and systems, which comprise glass and which use aptamers and in particular are battery-free, as well as methods for manufacturing or for operating the devices / devices, devices and systems.

Description

TECHNICAL FIELD

[0001] The present invention relates to a biosensor device, an implantable apparatus and a system comprising the device and / or the implantable apparatus, and to a method for manufacturing or for operating the apparatus / device, the apparatus and the system. BACKGROUND

[0002] A biosensor is a device that combines a biological component with a physico-chemical detector, or correspondingly a bio-chemical sensor, to detect and analyze specific biological or bio-chemical analytes (hereinafter: biomarkers or biological quantities). It is designed to convert a biological reaction, such as an interaction between a biological molecule and a target analyte, into a measurable signal.

[0003] The biological component of a biosensor can be a variety of biological elements, which include enzymes, antibodies, nucleic acids or whole cells. These components are selected based on their ability to recognize and interact with the target analyte of interest. When the target analyte binds to the biological component, it triggers a specific bio-chemical or bio-physical change.

[0004] The physico-chemical detector (herein also referred to as: biosensor device) is often referred to as a transducer, which converts the biological reaction into a measurable signal. This can be achieved through a variety of mechanisms, such as electrical, optical or electrochemical methods. The transducer generates a signal that is proportional to the concentration or presence of the target analyte, allowing for quantitative or qualitative analysis.

[0005] Biosensors have a wide range of applications in various fields, including healthcare, environmental monitoring, food safety and biotechnology. They can be used to detect and monitor biomarkers for disease diagnosis, measure glucose levels in diabetes management, detect environmental pollutants, or analyze DNA sequences. Biosensors have advantages such as rapid detection, sensitivity, selectivity, portability and real-time monitoring, which make them valuable tools in many industries.

[0006] The present invention particularly relates to an implantable apparatus for sensing at least one biological quantity within an animal or human body.

[0007] In this context, an implantable apparatus is an apparatus that is configured for implantation, in particular subcutaneous implantation, in particular complete implantation into an animal or human body.

[0008] An implantable apparatus for sensing a biological quantity within an animal or human body is also referred to as an implantable biosensor. An implantable biosensor is designed for monitoring and measuring a specific biological quantity, i.e. a biological parameter, within the body.

[0009] Implantable biosensors can be used for a wide range of applications, which include healthcare, biomedical research, veterinary. They can collect data and / or provide real-time data on a variety of biological quantities (i.e. physiological or biochemical parameters), enabling patients, healthcare professionals, and researchers to monitor conditions, diagnose diseases, and optimize treatment strategies.

[0010] Examples of implantable biosensors for sensing biological quantities include implantable glucose sensors, which are used to continuously monitor glucose levels in diabetic patients. They provide accurate and frequent glucose readings, which obviate the need for periodic finger-prick blood tests. Implantable pressure sensors, which are used to measure pressure at different parts of the body. For example, implantable intracranial pressure sensors can monitor pressure in the brain, which helps in diagnosing and managing conditions such as hydrocephalus or traumatic brain injury. Implantable temperature sensors, which are used to measure body temperature from within the body. They can be used for a variety of purposes, which include monitoring core body temperature during surgical procedures or tracking temperature changes in specific tissues. Implantable pH sensors, which measure the acidity or alkalinity of a solution, can be used to monitor pH levels in different body fluids or tissues, providing valuable information for diagnosing and managing conditions such as acidosis or alkalosis. Implantable oxygen sensors, which measure oxygen levels in body tissues or fluids. They can be used to monitor tissue oxygenation in intensive care settings, assess wound healing, or optimize oxygen delivery during surgical procedures.

[0011] There are also wearable and implantable biosensors that are not directly related to healthcare. These types of biosensors are designed for a variety of purposes outside of traditional healthcare. Implantable biosensors can be used to monitor physiological parameters of athletes during training and competition. These sensors can track indicators such as heart rate, body temperature, hydration levels, and lactate levels. The data collected can help athletes optimize their training, prevent overtraining, and improve athletic performance. Another application involves biohacking and quantified self: some people engage in the biohacking or quantified self movement, which involves tracking and optimizing various aspects of their own physiology. Implantable biosensors can be used to monitor parameters such as body temperature, sleep patterns, hormone levels, or even brain activity. This data can provide insights into personal health, habits, and performance optimization. Another application involves industrial monitoring. In industrial settings, implantable biosensors can be used for worker safety and environmental monitoring. For example, sensors implanted in workers can monitor their exposure to toxic substances, radiation levels, or other occupational hazards. This helps ensure a safe working environment and enables early detection of potential health risks. Another application involves animal tracking and research. Implantable biosensors are also used in wildlife and animal research for tracking, monitoring behavior, and collecting physiological data. These sensors can be implanted in animals to track migration patterns, habitat use, reproductive behavior, or monitor specific biological parameters. The data collected helps conservation efforts, understanding animal behavior, and ecological research. While these examples include implantable biosensors for non-healthcare purposes, they still involve monitoring and collecting biological / physiological quantities that include physical parameters directly related to biology, such as radiation exposure, etc. The applications can vary, but the fundamental principle of using biosensors to collect biological data, especially real-time data, remains consistent.

[0012] Implantable biosensors offer the advantage of simplifying monitoring, especially continuous real-time monitoring of biological quantities, which allows for early detection of abnormalities and timely intervention. Due to their small size and biocompatibility, they are designed to minimize tissue damage or discomfort. Additionally, they can utilize wireless communication technology to transmit data to external devices for analysis and interpretation.

[0013] The number of people accessing or using assisted reproductive technology (ART) is far from adequate in the global population. The European Society of Human Reproduction and Embryology estimates that the optimal utilization of in vitro fertilization (IVF) / intracytoplasmic sperm injection (ICSI) is 1,500 couples per million population per year (Andersen AN, et al; European IVF-monitoring programme (EIM), European Society of Human Reproduction and Embryology (ESHRE). Assisted reproductive technology in Europe, 2001. Results generated from European registers by ESHRE. Hum Reprod. 2005 May;20(5):1158-76. doi: 10.1093 / humrep / deh755. Epub 2005 Jan 21. PMID: 15665021.), requiring multiple ART cycles to achieve success. However, a 2011 study found that the total global utilization rate was only 477 cycles per million population, with an estimated 2 million ART cycles resulting in 500,000 babies (Adamson GD, et al, V. International Committee for Monitoring Assisted Reproductive Technology: World Report on Assisted Reproductive Technology, 2018.).

[0014] The low success rate of IVF is believed to be a result of implanting IVF-derived embryos into a non-receptive uterus. One reason for this low success rate can be the high levels of estrogen resulting from the administration of gonadotropins to retrieve multiple oocytes, rendering the uterus refractory. Thus, uterine receptivity, orchestrated by the interplay of progesterone and estrogen, is critical for successful implantation and pregnancy outcome. The results of experiments performed by Ma et al. using a mouse delayed implantation model provide evidence that a very narrow range of estrogen levels is a critical determinant in the transformation of uterine receptivity into a refractory state, which indicates that the uterus is extremely sensitive to estrogen levels associated with implantation (Ma WG, Song H, Das SK, Paria BC, Dey SK. Estrogen is a critical determinant that specifies the duration of the window of uterine receptivity for implantation. Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2963-8. doi: 10.1073 / pnas.0530162100. Epub 2003 Feb 24. PMID: 12601161; PMCID: PMC151449).

[0015] Furthermore, ART has multiple risks for women, such as ovarian hyperstimulation and pregnancy. In Denmark, 1.2% of women suffer from ovarian hyperstimulation syndrome (OHSS), with 75% of patients being hospitalized for more than 24 hours. This makes the industry improve precision in its treatments. A study of 2,700 women showed that individualized ovarian stimulation showed better results. However, reliable and accurate biomarkers are not often collected (Roudebush, W.E., Kivens, W.J., and Mattke, J.M. (2008) “Biomarkers of ovarian reserve,” Biomarker Insights, 2008 Apr 16; 3:259-268. doi: 10.4137 / bmi.s537. PMID: 19578510; PMCID: PMC2688347.). The procedures between clinics are not standardized, with some clinics advocating the use of blood markers for monitoring and others relying entirely on ultrasound scans, with differences still existing between the two.

[0016] Currently, IVF treatment requires women to have blood analysis every two days. This causes a great inconvenience; however, it is necessary in order to obtain more accurate results and efficacy of the treatment program and to adjust the clinical protocol during the treatment to increase the safety of the treatment. Women are also using other techniques to track hormone levels, such as using urine test paper to measure the presence of luteinizing hormone (LH), capturing saliva to measure progesterone levels and taking temperature with a wearable device to prove the ovulation cycle. Depending on the progress of the woman, women are monitored up to 2-3 times a week, sometimes more frequently. However, there are few alternatives to accurately monitor hormone levels and these are not done in real time. The lack of more precise data also limits precision medicine in the optimization of ART cycles and fertility. The inability to know hormone levels in women in real time for long periods of time can be revolutionary for the female population, not only in the ART field, but also in many other issues related to fertility, such as assisted fertilization or detection of specific ovarian conditions. By measuring the levels of estradiol, progesterone and luteinizing hormone in real time; the fertile window can be easily and accurately identified. Measuring estradiol and progesterone are the most reliable markers of ovarian events, as they are both direct markers from the ovary (Blackwell, L., Cooke, D., and Brown, S. (2018) “Self-Monitoring of Fertility Hormones: A New Era for Natural Family Planning?” Linacre Q. 2018 Feb; 85(1): 26-34. doi: 10.1177 / 0024363918756387. Epub 2018 Mar 28. PMID: 29970935; PMCID: PMC6027114).

[0017] For IVF, baseline hormone levels can be used to determine a woman’s natural fertility and to decide on a specific ovarian stimulation method. The technique can also monitor an individual’s hormonal response and determine the appropriate drug dosage and administration date. One example of this is the administration of human chorionic gonadotropin (hCG) on day 6 when serum estradiol levels are consistently elevated (Liu Y, Li J, Zhang W, Guo Y. Association between serum estradiol level on the hCG administration day and neonatal birthweight after IVF-ET among 3659 singleton live births. Sci Rep. 2021 Mar 16;11(1):6084. doi: 10.1038 / s41598-021-85692-7. PMID: 33727635; PMCID: PMC7966761).

[0018] Furthermore, the assessment of LH pulsatility is important for the clinical diagnosis of reproductive disorders, but current methods are hindered by frequent blood sampling and expensive continuous immunochemical analysis. Normal reproductive function is controlled by a highly coordinated hormonal feedback pattern of the entire hypothalamic-pituitary-gonadal (HPG) axis (Barbieri, R. L. The endocrinology of the menstrual cycle. Methods Mol. Biol. 1154, 145-169 (2014) doi: 10.1007 / 978-1-4939-0659-8_7. PMID: 24782009). The pulsatile release of LH, estradiol and progesterone is an essential element for the downstream regulation of sex steroid hormone synthesis and mature oocyte production. Changes in the hormone pulsatile secretion pattern are associated with hypothalamic dysfunction, which leads to many reproductive disorders, including polycystic ovary syndrome (PCOS) (Bachelot, A. et al. Luteinizing hormone pulsatility in patients with major ovarian hyperandrogenism. J. Endocrinol. Invest. 30, 636-646 (2007) doi: 10.1007 / BF03347443. PMID: 17923794), hypothalamic amenorrhea (Touraine, P. et al. Resumption of luteinizing hormone pulsatility and hypogonadotropic hypogonadism after endoscopic ventriculocisternostomy in a hydrocephalic patient. Fertil. Steril. 76, 390-393 (2001)) and delayed or precocious puberty. It is not feasible to measure hormone pulsatility in current clinical practice to determine altered secretion patterns, as this is extremely resource-consuming. Peripheral blood sampling has to be performed every 10 min for at least 8 h and continuous analysis by immunochemical analysis is expensive.However, human studies from professional clinical research groups (Prague, J. K. et al. Neurokinin 3 receptor antagonism as a novel treatment for menopausal hot flushes: a phase 2, randomised, double-blind, placebo controlled trial. Lancet 389, 1809-1820 (2017), doi: 10.1016 / S0140-6736(17)30823-1. Epub 2017 Apr 3. PMID: 28385352; PMCID: PMC5439024. Dhillo, W. S. et al. Kisspeptin-54 stimulates the hypothalamic-pituitary gonadal axis in human males. J. Clin. Endocrinol. Metab. 90, 6609-6615 (2005) Epub 2005 Sep 20. PMID: 16174713.) have shown that the ability to more accurately monitor hormonal concentration fluctuations has potential therapeutic benefits for women undergoing IVF and menopause. There are three major issues currently preventing widespread clinical hormonal concentration curve measurement and its pulsatility analysis: (1) resolution of hormonal concentration curves is limited by sampling protocols and immunochemical assays - there is no method to monitor LH pulsatility in real time. (2) measurement of hormonal concentration curves is cost-limited when using continuous clinical chemistry luminescence immunoassays (£20 per sample, 50 samples are needed for one patient). (3) hormonal pulsatility analysis is challenging as it usually requires advanced algorithms that can properly and effectively account for inherent biological variability, pulse-to-pulse variability, and physiological factors affecting hormone secretion and calculations, including clearance. There is a clear unmet medical need for better translational technologies that enable routine clinical LH pulsatility analysis in patients with reproductive disorders.

[0019] In particular, a biosensor device is a device that uses a biological component (such as an enzyme, antibody, or living cell) to detect the presence or concentration of a specific substance (such as a chemical or biological molecule). Biosensors generally include three components: a biological recognition element, a transducer, and a signal processing system.

[0020] A biological recognition element is the part of a biosensor that interacts with the target molecule and generates a signal. For example, an enzyme biosensor can use an enzyme to catalyze a reaction that produces a detectable product, while an antibody biosensor can use an antibody to bind a specific antigen and generate a signal. A transducer is the part of a biosensor that converts the signal generated by the biological recognition element into a measurable output signal, such as an electrical, optical, or acoustic signal. A signal processing system is the part of a biosensor that interprets the output signal and provides information about the concentration or presence of the target molecule. According to a preferred aspect of the present invention, the biological recognition element is an aptamer, in particular a biosensor membrane containing a layer of aptamers.

[0021] In vivo monitoring provides a direct link to the body processes performed at the cellular and tissue level. Implantable biosensors will be able to accurately detect local levels of different biomarkers.

[0022] Implantable devices have found widespread use in the field of contraception, where intrauterine devices have been on the market since the 19th century and have been widely marketed since the 1970s. In addition, a subcutaneous contraceptive implant was introduced in the United States in 2006. Although previous implantable devices have been used in this market, biosensor devices, in particular subcutaneous biosensor devices, have never been used for the treatment of infertility.

[0023] Therefore, there is a need for improved technology that is able to detect, in particular in real time, hormones. SUMMARY

[0024] In one aspect, the present invention relates to a biosensor device for sensing at least one hormone, comprising:

[0025] (a) at least one metal electrode surface; and

[0026] (b) a biosensor membrane comprising at least one aptamer attached to the metal electrode surface, wherein the aptamer:

[0027] (i) is capable of binding at least one hormone; and

[0028] (ii) is modified with one or more functional groups for attaching the at least one aptamer to the metal electrode surface.

[0029] Preferably, the biosensor device is configured for sensing at least one hormone in a body fluid, in particular in interstitial fluid.

[0030] Preferably, the biosensor device is an implantable device configured to be fully implantable into an animal or human body, in particular subcutaneously. Preferably, the biosensor device is a wearable device, in particular configured to be used in contact with the skin of an animal or human.

[0031] Preferably, the at least one hormone is selected from the group consisting of estradiol, luteinizing hormone (LH), progesterone, and any combination thereof.

[0032] Preferably, the biosensor device further comprises a carrier, in particular a biocompatible glass carrier, which is preferably transparent, or preferably non-transparent.

[0033] Preferably, the aptamer is a single-stranded nucleic acid molecule, and / or is preferably a DNA or RNA molecule, which binds specifically with high affinity and specificity.

[0034] Preferably, the aptamer has a length of about 25 to 70 nucleotides, preferably about 30 to about 65 nucleotides.

[0035] Preferably, the functional group for attaching the at least one aptamer to the metal electrode surface is a thiol group.

[0036] Preferably, the functional group for attaching the at least one aptamer to the metal electrode surface is present at the end of the aptamer.

[0037] Preferably, the aptamer is a single-stranded nucleic acid molecule, preferably a DNA molecule, and the functional group for attaching the at least one aptamer to the metal electrode surface is present at the 3’- or 5’-end, preferably the 5’-end.

[0038] Preferably, the aptamer is capable of binding at least one hormone selected from the group consisting of estradiol, luteinizing hormone (LH), and progesterone.

[0039] Preferably, the aptamer is one or more selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4.

[0040] Further, the present application relates to a biosensor device according to the present application for use in diagnostics, in particular for sensing at least one hormone, more particularly for monitoring hormones during assisted reproductive technology (ART), during menopause, in hormonal disorders such as polycystic ovary syndrome (PCOS), in endometriosis, and / or during a course of hormone therapy.

[0041] Further, the present application relates to the use of a biosensor device according to the present application for in vitro diagnostics, in particular for sensing at least one hormone.

[0042] Further, the present application relates to an implantable device for implantation, in particular subcutaneous implantation, into an animal or human body, comprising

[0043] - A biosensor device according to the present invention, and

[0044] - An electronic control device for controlling the biosensor device, particularly for generating, collecting, and preferably encrypting measurement data obtained through sensing by the biosensor device.

[0045] Furthermore, the present invention relates to a system for sensing at least one hormone, comprising:

[0046] - The implantable device according to the invention further includes a communication means for wireless data communication with an external computer device, particularly an NFC (Near Field Communication) device, which is separate from the implantable device, and

[0047] - The external computer device includes a communication device for wireless data communication with an implantable device, particularly an NFC device.

[0048] Furthermore, the present invention relates to a method for operating a biosensor device according to any one of the preceding claims, comprising the following steps:

[0049] (a) Contact the surface of the metal electrode with body fluid, particularly interstitial fluid;

[0050] (b) binding at least one hormone to at least one aptamer on the surface of the metal electrode; and

[0051] (c) The binding of at least one hormone is detected by electrically controlling at least one metal electrode surface, preferably by differential pulse voltammetry (DPV). Detailed Implementation

[0052] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0053] It is worth noting that the use of the undefined article "a (a or an)" means "one or more". Therefore, for example, the term "a hormone" includes both "one" and "more than one" hormones.

[0054] As used herein, the term "comprising" means "including but not limited to". This term is intended to be open-ended to specify the presence of any of the stated features, elements, integers, steps, or components, but does not preclude the addition of one or more other features, elements, integers, steps, components, or combinations thereof. Therefore, the term "comprising" includes the more restrictive terms "consisting of" and "substantially consisting of". In one embodiment, as used throughout this application, particularly in the claims, the term "comprising" may be replaced by the terms "consisting of" or "substantially consisting of".

[0055] According to a preferred aspect, the present invention relates to a biosensor device for sensing at least one hormone, comprising at least one metal electrode surface and a biosensor membrane, the biosensor membrane comprising at least one aptamer attached to the metal electrode surface.

[0056] The biosensor device according to the present invention improves IVF technology and the way treatment is administered today because it enables real-time monitoring. It has also led to innovations in other areas of women's health where frequent and accurate hormone level capture is important, such as polycystic ovary syndrome, endometriosis, and menopause. Applications in these areas will help clinicians make earlier diagnoses and provide more accurate treatments. Furthermore, data collected using the biosensor of the present invention can be used to build better predictive and personalized machine learning algorithms, with improved capabilities and more accurate results that are crucial in clinical practice.

[0057] The biosensing membrane (also known as a "biosensor membrane") of a biosensor device is also referred to as an "aptasensor." The aptasensor is capable of accurately and repeatedly detecting a target analyte, particularly a biomass. The aptasensor is preferably prepared via an electrochemical process using several steps as described in the embodiments herein. These steps preferably include: activating the surface of a metal electrode serving as the working electrode; preferably: adding one or more nanoparticle layers to increase sensitivity; adding a functional sensing layer formed of the aptas; and preferably: finally adding a protective porous biocompatible membrane. When the biosensor comes into contact with even trace amounts of the analyte, the aptasensor binds to it, and the measurement signal is sensed and recorded. Using this method, precise quantities, particularly the amount or concentration of the analyte, or changes in the analyte concentration, can be measured, especially time-dependent changes. Preferably, the aptasensor is calibrated and / or otherwise validated prior to deployment.

[0058] In principle, biosensing of analytes can be achieved using antibodies, aptamers, and protein receptors via electrochemical sensors. The advantage of aptamers is that they can bind to the target analyte in a reproducible manner, while antibody binding to the target is irreversible. Compared to other biosensors using enzymes or antibodies, the chemical simplicity, stability, specificity, and selectivity of aptamers enable continuous in vivo monitoring. Their advantages (including equal or higher affinity and specificity to the target, smaller size, easier modification and immobilization, better stability, and higher reproducibility) allow for the development of the highly specific biosensors of this invention. The inventors have discovered that the use of aptamers provides significant and unexpected advantages in the lifetime of biosensor devices, surpassing other biorecognition elements used for in vivo detection of biomarkers (particularly hormones).

[0059] Integrating aptamers into microelectrodes small enough for implantation presents challenges, such as limited current signals due to low aptamer density on the electrode surface. This, in turn, leads to a low signal-to-noise ratio (SNR), which is exacerbated in embodiments of the invention by limitations of low-power communication devices, particularly NFC devices. To improve SNR, the inventors have developed a strategy involving the use of nanoparticles, particularly gold and / or platinum nanoparticles, carbon nanotubes, or carbon nanoparticles. The use of such nanoparticles significantly increases the total sensing surface area and improves SNR. Incorporating nanoparticles into the sensing layer significantly enhances the sensitivity of electrochemical sensors. These nanoparticles are used for a variety of purposes, such as modifying electrode surfaces, labeling specific target molecules for detection, amplifying signals, and acting as catalysts for ongoing chemical reactions. Furthermore, nanoparticles offer excellent biocompatibility and provide a larger surface area on the electrode, thereby improving electron transfer capabilities, facilitating the immobilization of bioactive substances on the electrode surface, and reducing the detection time. The nanoparticles used can vary in size (in the nanometer range), shape (spherical, cylindrical, planar, etc.), and composition, including noble metal nanomaterials (such as gold and silver), semiconductor materials (such as quantum dots), carbon-based nanomaterials (such as carbon nanotubes and graphene oxide), and composite nanomaterials. Depending on the specific nanomaterial and substrate surface, various methods can be used to deposit nanoparticles, such as dip coating, spin coating, solvent evaporation, chemical vapor deposition, and transfer printing.

[0060] Preferably, the biosensor device is configured to sense at least one hormone in physiological fluids or bodily fluids (e.g., blood, serum, saliva, urine, sweat, tears, mucus and vaginal secretions, uterine wall and menstrual blood) or interstitial fluid, preferably: interstitial fluid.

[0061] As described above, in vivo monitoring provides a direct connection to bodily processes performed at the cellular and tissue levels. Implantable biosensors would be able to accurately detect local levels of various biomarkers. Therefore, in one embodiment, the biosensor device is an implantable device configured to be fully implantable into an animal or human, particularly subcutaneously.

[0062] As used herein, "aptamer" refers to a short, single-stranded ribonucleic acid (RNA), deoxyribonucleic acid (DNA), or xenobiotic nucleic acid (XNA) that binds to a specific target molecule with affinity. High affinity is primarily achieved through complex and diverse secondary and tertiary structures resulting from the intramolecular doubling of the single-stranded nucleic acid, which produces a defined three-dimensional structure. Typically, aptamers are approximately 20 to 100 nucleotides in size. Target molecules can include small molecules, heavy metal ions, larger ligands (e.g., proteins), and even intact cells. Aptamers can also be modified with specific functional groups to enhance their function and / or compatibility with larger engineered molecular systems. Aptamers have applications in a wide range of fields, including but not limited to sensing, therapeutics, reagent production, and engineering. For example, aptamers are used as sensing molecules to bind to specific biomolecules (e.g., hormones) present in body fluids, particularly interstitial fluid.

[0063] In the context of this invention, any aptamer capable of binding to the hormone to be sensed can be used. Those skilled in the art know how to develop aptamers that specifically bind to a given hormone, including but not limited to evolution through exponentially enriched ligand systems (SELEX). SELEX is a multi-step process designed to identify and optimize short, single-stranded DNA or RNA molecules—i.e., aptamers—that can bind to a specific target molecule with high affinity and specificity. SELEX methods are known to those skilled in the art and can vary slightly depending on the specific target, the target molecule, and the resources available to the researcher.

[0064] For example, the SELEX method may include the following steps:

[0065] 1) Target recognition: The first step is to identify the target molecule that the aptamer will bind to. This can be a small molecule, a protein, or even an intact cell.

[0066] 2) Library generation: Synthesizing large libraries of random DNA or RNA sequences, which typically contain billions of different sequences. These sequences serve as the starting point for aptamer selection.

[0067] 3) Selection: Begin with the true SELEX method. Incubate the library of random sequences with the target molecule to allow the aptamer with binding affinity for the target to bind.

[0068] 4) Partitioning: Separating bound aptamers from unbound aptamers. Various techniques (e.g., filtration, affinity chromatography, or magnetic separation) can be used for this purpose and are known to those skilled in the art.

[0069] 5) Amplification: The bound aptamers are eluted and amplified using a method called polymerase chain reaction (PCR) or reverse transcription PCR (RT-PCR). This step increases the concentration of the selected aptamers for subsequent rounds of selection.

[0070] 6) Iterative multiple rounds: Repeat steps 3-5 times, typically 8-15 times, to enrich a library of aptamers with high affinity and specificity for the target. The stringency of the selection criteria gradually increases in each round to facilitate the selection of the optimal aptamer.

[0071] 7) Sequencing and Analysis: After the required number of selection rounds, the enriched aptamer library is sequenced to determine the DNA or RNA sequences of the selected aptamers. Bioinformatics analysis is then performed to identify common motifs or structural features among the aptamers.

[0072] 8) Aptamer optimization: Selected aptamer sequences are synthesized and further optimized to improve their binding performance, stability, and specificity. This can be achieved through chemical modification, truncation, or mutagenesis.

[0073] 9) Validation and characterization: Optimized aptamers are tested using a variety of techniques (e.g., surface plasmon resonance, fluorescence assays, or cell-based assays) to confirm their binding affinity, specificity, and functionality. These methods are known to those skilled in the art.

[0074] 10) Applications: Finally, validated aptamers can be used in a wide range of applications, including diagnostics, therapy, biosensing, drug delivery, and biomarker discovery.

[0075] Preferably, the aptamer is a single-stranded nucleic acid molecule, more preferably a DNA molecule. For example, the aptamer is an oligonucleotide.

[0076] Preferably, the aptamer has a length of about 25 to 70 nucleotides, more preferably about 30 to about 65 nucleotides.

[0077] As described herein, aptamers can be modified with functional groups to enhance their functionality. The methods for modifying aptamers with functional groups can vary and generally depend on the specific functional group and / or the attachment site of the functional group on the aptamer. These methods involve fundamental chemistry and are well known in the art. In one embodiment, the aptamer is modified with a functional group for attaching at least one aptamer to a metal electrode surface. In another embodiment, the aptamer is modified with one or more functional groups for attaching at least one aptamer to a metal electrode surface. In principle, any functional group capable of enabling the aptamer to attach to the metal electrode surface can be used, and the functional group can depend on the nature and composition of the metal electrode surface. In a particular embodiment, the functional group is a thiol group (-SH). The thiol group on the aptamer enables the aptamer to attach to the metal electrode surface, particularly a gold electrode surface. This is referred to as a self-assembled monolayer (SAM). In principle, the functional group can be present at any site on the aptamer, provided that the binding activity of the aptamer to the target analyte is not weakened or impaired. The exact location of the functional group can depend on the specific aptamer and the surface of the metal electrode.

[0078] A "self-assembled monolayer" (SAM) is a monolayer of molecules that spontaneously organizes into an ordered pattern on a solid substrate. Unbound by any specific theory, this organization occurs through intermolecular forces, such as van der Waals interactions, hydrogen bonds, or electrostatic interactions. SAMs are typically formed by immersing the substrate in a solution containing the desired molecules, allowing them to adsorb and self-assemble on the surface. SAMs play a crucial role in the crosslinking process by providing a template for the attachment of crosslinking agents or polymers. The functional groups present on the SAM surface can react with crosslinking agents, resulting in the formation of covalent bonds between molecules. This crosslinking process enhances the stability and mechanical properties of the resulting material. SAMs provide a well-defined and controllable surface structure that can be used for crosslinking reactions, enabling the development of advanced materials with customized properties and improved performance.

[0079] Preferably, the functional group for attaching at least one aptamer to the surface of the metal electrode is located at the end of the aptamer. For example, the aptamer is a single-stranded nucleic acid molecule, preferably a DNA molecule. Then, the functional group for attaching at least one aptamer to the surface of the metal electrode can be located at the 3' end and / or the 5' end. Preferably, the aptamer is a single-stranded nucleic acid molecule, preferably a DNA molecule, and the functional group for attaching at least one aptamer to the surface of the metal electrode is located at the 3' end. Preferably, the aptamer is a single-stranded nucleic acid molecule, preferably a DNA molecule, and the functional group for attaching at least one aptamer to the surface of the metal electrode is located at the 5' end. Attachment at the 5' end can be achieved, for example, by replacing the hydroxyl group of the phosphorus group of the 5' terminal nucleotide with a thiol group. Attachment at the 3' end can be achieved, for example, by replacing the hydroxyl group of the ribosome unit of the 3' terminal nucleotide with a thiol group.

[0080] Preferably, the biosensor of the present invention is configured to sense at least one hormone. Therefore, the aptamer is capable of binding to at least one hormone. The hormone to be detected includes, but is not limited to, estradiol, luteinizing hormone (LH), and progesterone. Preferably, the hormone to be detected is selected from the group consisting of: estradiol, luteinizing hormone (LH), progesterone, and any combination thereof. Furthermore, the hormone to be detected can be selected from the group consisting of: estradiol, luteinizing hormone (LH), progesterone, cortisol, FSH, testosterone, serotonin, triiodothyronine (T3), tetraiodothyronine (T4), calcitonin, insulin, melatonin, TSH, hGH, AmH, glucagon, and any combination thereof.

[0081] Therefore, preferably, the aptamer is capable of binding at least one hormone selected from the group consisting of estradiol, luteinizing hormone, and progesterone. For example, a biosensor is used to sense estradiol. Therefore, the biosensor membrane contains at least one aptamer capable of binding estradiol. For example, a biosensor is used to sense luteinizing hormone. Therefore, the biosensor membrane contains at least one aptamer capable of binding luteinizing hormone. For example, a biosensor is used to sense progesterone. Therefore, the biosensor membrane contains at least one aptamer capable of binding progesterone. In another example, a biosensor is used to sense, for example, a combination of estradiol and progesterone. Therefore, the biosensor membrane contains at least one aptamer capable of binding estradiol and at least one aptamer capable of binding progesterone. In another example, a biosensor is used to sense, for example, a combination of estradiol, luteinizing hormone, and progesterone. Therefore, the biosensor membrane contains at least one aptamer capable of binding estradiol, at least one aptamer capable of binding luteinizing hormone, and at least one aptamer capable of binding progesterone. Preferably, the aptamer of the biosensor membrane comprises at least one aptamer to be sensed or a combination of aptamers to be sensed. Preferably, the biosensor membrane comprises a plurality of aptamers that bind to a specific hormone to be sensed. In one embodiment, the plurality of aptamers to be sensed are arranged in a layer on the surface of a metal electrode.

[0082] The amount of one or more individual aptamers capable of binding specific hormones, or the relative amount of aptamers capable of binding different hormones, present on a biosensor membrane is variable and can depend on the desired stability and mechanical properties of the resulting material as well as the desired structure. This enables the development of advanced materials with customized properties and improved performance.

[0083] Preferably, the aptamer is one or more selected from the group consisting of:

[0084] LH aptamers:

[0085] 5'-TATGGTATGCTGTGTGGTATGGGGTGGCGTGCTCT-3' (SEQ ID NO: 1)

[0086] Preferably, the functional group used to attach at least one aptamer to the surface of the metal electrode is a thiol group, and may be present at the 3' end and / or 5' end of SEQ ID NO: 1. In a preferred embodiment, the functional group used to attach at least one aptamer to the surface of the metal electrode is a thiol group, and is present at the 5' end.

[0087] β-estradiol aptamers:

[0088] 5'-TTTTTTTTTTTTTTTGCTTCCAGCTTATTGAATTACACGCAGAGGGTA-3' (SEQ ID NO: 2)

[0089] Preferably, the functional group used to attach at least one aptamer to the surface of the metal electrode is a thiol group, and may be present at the 3' end and / or 5' end of SEQ ID NO: 2. Preferably, the functional group used to attach at least one aptamer to the surface of the metal electrode is a thiol group, and is present at the 5' end.

[0090] 5'-GCGGCTCTGCGCATTCAATTGCTGCGCGCTGAAGCGCGGAAGCTTTTTTTTTTT-3' (SEQ IDNO: 3)

[0091] Preferably, the functional group used to attach at least one aptamer to the surface of the metal electrode is a thiol group, and may be present at the 3' end and / or 5' end of SEQ ID NO: 3. In a preferred embodiment, the functional group used to attach at least one aptamer to the surface of the metal electrode is a thiol group, and is present at the 3' end.

[0092] Progesterone aptamers:

[0093] 5'-GCATCACACACCGATACTCACCCGCCTGATTAACATTAGCC CACCGCCCACCCCCGCTGC-3' (SEQ ID NO: 4)

[0094] Preferably, the functional group used to attach at least one aptamer to the surface of the metal electrode is a thiol group, and may be present at the 3' end and / or 5' end of SEQ ID NO: 4. Preferably, the functional group used to attach at least one aptamer to the surface of the metal electrode is a thiol group, and is present at the 5' end.

[0095] Preferably, the present invention relates to a biosensor device for diagnostic purposes. Preferably, the biosensor device of the present invention is used to sense hormones. For example, the biosensor can be used to determine at least the nature of a hormone and / or the concentration of said hormone. Hormone sensing can be used to monitor hormones during assisted reproductive technology (ART) or menopause, or to monitor the menstrual cycle. Hormone sensing can also be used to study the progression or treatment of hormonal disorders, or to diagnose hormonal disorders such as polycystic ovary syndrome (PCOS) or endometriosis. Hormone sensing can also be useful during sex reassignment, menopause, and hormone therapy during ART.

[0096] Preferably, the present invention relates to in vitro diagnostics, particularly for sensing at least one hormone. For example, in vitro diagnostics can be used to diagnose hormones from any sample derived from a subject (e.g., a human or animal), said sample including but not limited to bodily fluids, particularly blood, serum, urine, interstitial fluid, or saliva. In vitro diagnostics can also be useful in basic research, for example, for calibrating biosensors (see also embodiments herein).

[0097] Furthermore, the present invention relates to a method for operating a biosensor device according to any one of the preceding claims, comprising the following steps:

[0098] (a) Contact the biosensor membrane with body fluids, especially interstitial fluid;

[0099] (b) binding at least one hormone to at least one aptamer of the biosensor membrane; and

[0100] (c) Detecting the binding of the at least one hormone by electrically controlling at least one metal electrode surface.

[0101] Preferably, step (b) is performed under physiological conditions.

[0102] Preferably, step (c) is performed using differential pulse voltammetry (DPV). DPV is an electrochemical technique for signal detection and quantification in a variety of applications. It involves applying a series of voltage pulses to an electrochemical cell and measuring the resulting current response.

[0103] Preferably, the biosensor device includes at least one electrode, the electrode comprising a metal electrode surface supporting a biosensor membrane. Preferably, the biosensor device further includes a working electrode, preferably a reference electrode, and preferably a counter electrode. Preferably, the biosensor device includes an electronic control device for controlling at least one metal electrode surface. Preferably, the electronic control device is configured, and in particular programmed, to perform an electrical measurement of at least one biomass by controlling the at least one metal electrode surface. Preferably, the electrical measurement is a voltammetry method, preferably a DPV (differential pulse voltammetry) measurement. DPV measurement will be explained in more detail below.

[0104] The present invention also relates to a method of treating a subject (e.g., a human or animal) with a hormonal disorder using the biosensor device or implantable device or system of the present invention. Hormone sensing in treatment scenarios can be useful during assisted reproductive technology (ART) or during menopause, or for monitoring disrupted menstrual cycles. Hormone sensing can also be useful in scenarios treating hormonal disorders (e.g., polycystic ovary syndrome (PCOS) or endometriosis). Hormone sensing can also be useful in degenerative scenarios. For example, the biosensor can be used to determine at least the nature of the hormone and / or the concentration of said hormone, and based on the measurement results, hormone replacement therapy can be prescribed for the patient. In one embodiment, the method includes treating a hormonal disorder (e.g., infertility) and includes the following steps:

[0105] (a) Contact the biosensing membrane with the subject’s body fluids, particularly interstitial fluid;

[0106] (b) To bind at least one hormone to at least one aptamer of a biosensor membrane;

[0107] (c) Detecting the binding of the at least one hormone by electrically controlling the surface of at least one metal electrode, particularly by voltammetry, such as DPV;

[0108] (d) Determine the nature and / or concentration of the at least one hormone;

[0109] (e) Treat the subject with progesterone if the sensed progesterone level is ≥ a threshold level, preferably 1.5 ng / mL; and / or treat the subject with hCG if the sensed estradiol level is in the range of about 1,000-2,000 pg / mL.

[0110] Example 1

[0111] The following steps describe a method for preparing aptamers prior to self-assembly coating on the electrode surface:

[0112] 1) The lyophilized powder vials were centrifuged to obtain dried aptamer pellets in the vials (aptamers for thiol modification of estradiol, LH and progesterone obtained from Basepair Bio Company, the sequences of which are shown in SEQ ID NO: 1 to 4).

[0113] 2) Resuspend the aptamer precipitate in nuclease-free water with 10 mM Tris, pH 7.5, 0.1 mM EDTA (0.6057 g Tris, 0.0184 g EDTA, 500 ml DI water; resuspension buffer).

[0114] 3) Add resuspension buffer to the aptamer precipitate to achieve a concentration of 100 µM.

[0115] 4) Prepare folding buffer (1 mM MgCl2, 1x PBS, pH 7.5) and dilute the aptamer solution to 10x working concentration in the folding buffer. Heat the solution to 90-95°C for 5 minutes.

[0116] 5) Allow the solution to cool to room temperature for 15 minutes.

[0117] 6) Prepare reducing DTT buffer (20 mM DTT in 1x Te buffer (10 mM Tris, pH 8; 1 mM EDTA) or 20 mM DTT in Tris-HCl buffer, 1 M dithiothreitol (DTT)). This step should be completed immediately before use.

[0118] 7) Reduce the thiol-modified aptamer with 20 mM DTT in 1x TE buffer, and incubate with an equal volume of folded aptamer (1:1) and aptamer reduction buffer at room temperature (20-25°C) for 1 hour.

[0119] 8) Use a desalting column or buffer exchange column to remove DTT.

[0120] 9) Dilute the aptamer to the final working concentration using a buffer containing 1 mM MgCl2 (in nuclease-free water PBS).

[0121] 10) The aptamer is ready for coating the electrode.

[0122] Example 2

[0123] The following steps describe a method for preparing a functionalized gold electrode for hormone measurement:

[0124] 1) While preparing the aptamer (Example 1), a gold electrode on glass was prepared.

[0125] 2) The electrode was activated in 50 mM sulfuric acid using a three-electrode method (counter electrode, reference Ag / Cl electrode, and gold working electrode) and 45 segments.

[0126] 3) After this step, gently wash the sensor with 10 mM sulfuric acid.

[0127] 4) Record the DPV (differential pulse voltammetry) signal of the gold electrode, and then add the aptamer.

[0128] 5) Add 10 µl of aptamer solution to the gold working electrode to a concentration of 5 mM and incubate for 16 hours to immobilize the aptamer on the gold electrode by self-assembly of a monolayer.

[0129] 6) Then, use sterile PBS to wash away excess aptamers from the surface.

[0130] 7) Add 2 mM MCH (6-mercapto-1-hexanol) to the electrode as a blocking agent and incubate for 1 hour.

[0131] 8) Record the DPV signal during the addition of the hormone stock solution to the ISF (interstitial fluid) (from 1 pg / ml to 1 ug / ml).

[0132] 9) Finally, the peak current of each DPV signal is analyzed to measure sensitivity.

[0133] 10) Repeat the experiment at least three times, and the results are as follows: Figure 11 As shown.

[0134] According to a preferred aspect, the present invention relates to an implantable device / implantable biosensor device.

[0135] Implantable biosensors are at risk of rejection or adverse reactions due to a variety of factors, including the body’s immune response and the materials used in the implant.

[0136] When a foreign object is introduced into the body, the immune system recognizes it as a non-self substance and may initiate an immune response to eliminate or neutralize it. This immune response can manifest as inflammation, swelling, pain, or even immune-mediated implant rejection. The severity of the immune response varies from person to person and can be influenced by factors such as genetics and overall health. When using implantable biosensors, it is necessary to reduce the risk of severe immune responses. Therefore, implantable devices are made of biocompatible materials, meaning they are designed to minimize adverse effects on living tissue. The choice of materials used in implants plays a crucial role in determining their compatibility with the body. Some materials can trigger an immune response or cause adverse reactions more readily than others. For example, certain metals or polymers can cause allergic reactions or release toxic substances upon contact with bodily fluids or tissues.

[0137] Implantation procedures carry the risk of introducing bacteria or other microorganisms into the body, which can lead to infection. Infection can cause local inflammation, tissue damage, and even require implant removal. There is a need for implantable biosensors that minimize the risk of introducing bacteria or other microorganisms into the body.

[0138] The skill and technique of the surgeon performing the implantation procedure also affect the risk of adverse reactions. Improper implant placement, surgical trauma, or inadequate sterilization can increase the likelihood of complications and subsequent rejection. There is a need to provide implantable biosensors that are easy for surgeons to use.

[0139] Implantable devices involve significant financial investment, including the cost of the device itself, surgery, and post-implantation care. Therefore, maximizing the lifespan of implantable devices is crucial for optimizing the cost-effectiveness of treatment. A longer lifespan for implantable devices reduces the need for frequent replacements or repairs, thereby lowering overall healthcare costs.

[0140] Implantable devices are typically designed to provide long-term solutions for medical conditions or disabilities. The longer lifespan of implantable devices means longer periods of improved function, convenience, and quality of life for patients. Avoiding frequent replacements or maintenance procedures minimizes disruption to daily life and reduces the burden on patients and caregivers.

[0141] Implantable procedures inherently carry risks, including potential complications such as infection, bleeding, and tissue damage. Minimizing the need for additional surgeries or modifications due to implant failure or degradation reduces the cumulative risks associated with multiple surgeries. It also reduces the likelihood of additional complications and their associated medical costs.

[0142] Implantable devices often play a vital role in monitoring and managing medical conditions. For example, implantable sensors for glucose monitoring in diabetic patients provide valuable data for treatment adjustments. The lifespan of such devices ensures continuous and reliable data collection, enabling healthcare professionals to make informed decisions about treatment plans.

[0143] Implantable devices should be designed to maintain their functionality and biocompatibility throughout their intended lifespan to minimize the risk of complications, adverse reactions, or device failure. Regular monitoring, follow-up care, and periodic evaluation of implantable devices are necessary to ensure their continued effectiveness and safety.

[0144] Therefore, according to one aspect of the invention, the object of the invention is to overcome one or more of the problems of the above-mentioned implantable biosensors, and in particular to provide an implantable device with sufficient lifespan and providing long-term safety.

[0145] This objective is achieved by an implantable device according to a preferred aspect of the invention, which is an implantable device for sensing at least one biomass in an animal or human body, particularly for implantation, especially subcutaneous implantation, into an animal or human body, comprising a biosensor device for sensing biomass in vivo and an electronic control device for controlling said biosensor device, wherein the implantable device includes an encapsulation device for at least partially encapsulating at least one of the biosensor device and the electronic control device, and wherein said encapsulation device is at least partially made of glass.

[0146] The encapsulation device may be a single component or may include more than one encapsulation component forming the encapsulation device. Using multiple encapsulation components facilitates the assembly of the implantable device. Preferably, the encapsulation device includes two encapsulation components that together encapsulate a chamber for receiving electronic control devices and preferably other functional devices (e.g., communication devices or biosensor devices). Preferably, the encapsulation device specifically includes one, two, three, four, five, or six, or more, encapsulation components that together encapsulate the chamber. Preferably, at least one, two, three, four, five, or more of the encapsulation components comprise glass, or are partially or entirely made of glass, particularly substantially entirely made of glass. The glass is preferably transparent, or preferably opaque.

[0147] Preferably, the packaging device includes at least one wafer substrate comprising glass or being partially or entirely made of glass, wherein "entirely made of glass" includes cases where the wafer is particularly substantially entirely made of glass, meaning that it may include a low proportion of non-glass portions. The glass is preferably partially or completely transparent, and / or preferably partially or completely opaque. Preferably, the glass wafer includes a plurality of vias that are open or preferably filled with a non-glass material, particularly metal, and these vias are particularly cylindrical and may have a height of about 100 to 1000 micrometers, preferably 200 to 800 micrometers, preferably 300 to 700 micrometers, preferably 400 to 600 micrometers, preferably 500 micrometers, a diameter of preferably 20 to 500 micrometers, preferably 40 to 200 micrometers, preferably 50 to 150 micrometers, and a spacing of preferably 1000 to 20 micrometers, preferably 800 to 30 micrometers, preferably 500 to 50 micrometers, or preferably 300 to 50 micrometers. For example, HermeS® glass wafer substrates, sourced from Schott AG in Germany, are glass wafers made essentially of glass. They include hermetically sealed, solid through-glass vias (TGVs). Using such glass, finely pitched vias reliably conduct electrical signals and power into and out of the implantable device because the vias are solidly filled with conductive metal. Generally, a wafer substrate is understood to refer to a solid, planar plate or disk-shaped element used as a substrate for at least one electrode or other sensing element. Preferably, the wafer substrate is an element cut from a larger wafer, particularly a glass wafer.

[0148] Preferably, the wafer substrate contains at least one metal-filled via connecting to a device of an implantable device, particularly to any of the following devices: electronic control devices, biosensor devices (particularly at least one electrode of a biosensor device), or communication devices. Preferably, the wafer substrate contains at least one metal-filled via connecting to at least one sensing element (particularly at least one electrode) of a biosensor device on one side of the wafer substrate and to an electronic control device on the other side of the wafer substrate. Preferably, the wafer substrate contains at least one metal-filled via connecting to a communication device on one side of the wafer substrate and to an electronic control device on the other side of the wafer substrate.

[0149] A wafer substrate is understood as a thin, substantially planar, preferably substantially rectangular or substantially circular sheet of material. The wafer substrate preferably provides a solid and uniform surface, on which various electronic structures, circuits, or electrodes can be constructed, forming part of electronic control devices and / or communication devices.

[0150] The wafer substrate is preferably made of glass. It can also be made of another amorphous or crystalline material, such as a semiconductor material like silicon (Si). Glass wafers are preferred because they offer the desired electrical and mechanical properties, availability, and compatibility with standard photolithography processes. Furthermore, glass provides excellent biocompatibility even without a biocompatible coating; however, a biocompatible coating is also preferred for packaging devices that are partially or entirely composed of glass or non-glass materials, or correspondingly, for the wafer substrate. The coating may be a glass coating.

[0151] The inventors of this invention have discovered that glass can be used to manufacture safe and reliable encapsulation devices, which in particular contain one or more hermetic sealed chambers with a long lifespan.

[0152] The glass used for encapsulation can preferably be any one of bioactive glass, borosilicate glass, aluminosilicate glass, phosphate glass, silica glass, or chalcogenide glass. Bioactive glasses have the ability to bind to living tissue. They typically contain calcium, phosphorus, and silica as their main components. Borosilicate glasses (e.g., Pyrex) have high chemical resistance and thermal stability. Aluminosilicate glasses are a class of glasses containing aluminum and silicon. They have good mechanical strength. Phosphate glasses typically contain phosphorus pentoxide (P₂O₅) as their main component. They exhibit high solubility in aqueous solutions. Silica glasses (e.g., fused silica) are primarily composed of silica (SiO₂). They have good optical transparency, low thermal expansion, and good chemical stability. Chalcogenide glasses are composed of chalcogen elements (including sulfur, selenium, and tellurium). They possess unique optical and electrical properties. Preferably, the glass is any one of Borofloat® 33, AF 32® eco 33, or D 263® Teco, all purchased from Schott AG, Germany. Preferably, the encapsulation device is made of a type of glass material, which allows for easy assembly of the encapsulation device.

[0153] In a preferred aspect, the at least one wafer substrate carries one or more metal electrodes, which in particular form part of a biosensor device and preferably carry a biosensor membrane.

[0154] In a preferred aspect, the at least one wafer substrate carries an electronic control device, which is specifically used to control the measurement process using a biosensor device.

[0155] In a preferred aspect, the at least one wafer substrate carries a communication device, particularly an RFID device, and especially an NFC device.

[0156] In a preferred aspect, a wafer substrate carries two or all of the following devices: an electronic control device, at least one sensing element (particularly at least one electrode) of a biosensor device, and a communication device.

[0157] In a preferred aspect, the packaging device comprises two wafer substrates stacked on top of each other, particularly arranged in parallel, and preferably at a distance to encapsulate a chamber, which may in particular contain one or more of an electronic control device, a biosensor device, and a communication device. The distance may preferably be 200 to 5000 micrometers, more preferably 300 to 3000 micrometers, or more preferably 500 to 2500 micrometers. The sides of the chamber may be closed by rod-like members, which may be integral with either of the wafer substrates or may be separate components; preferably, the rod-like members are particularly joined to either of the substrate wafers by welding or adhesive bonding.

[0158] In a preferred aspect, at least two glass components of the encapsulation device, particularly two glass wafer substrates, are preferably arranged in a stacked manner and joined together by femtosecond laser splicing. Surprisingly, this technique has been found to ensure complete airtightness even under pressure conditions and during long-term test use of the implantable device, thereby protecting the electronic equipment and internal chambers from exposure to the aforementioned physiological fluids.

[0159] In a preferred aspect, the packaging device comprises a stack of two, three, four or more wafer substrates, which are preferably arranged at a distance to encapsulate the chamber.

[0160] The wafer substrate may include a flange portion that forms a groove-shaped substrate member. The hollow portion of such a groove can serve as a chamber for receiving or covering any of electronic control devices, biosensor devices, and communication devices.

[0161] The encapsulation device can be in capsule form during assembly. This facilitates implantation of the implantable device using, for example, a syringe.

[0162] The encapsulation device can be assembled in the form of a box, and in particular a cuboid with quadrilateral faces, preferably a rectangular prism. The edges and / or corners can be rounded, particularly with a corner radius of 0.05 to 5 mm, preferably 0.1 to 3 mm.

[0163] In a preferred aspect, the packaging device includes a main wafer substrate and a slotted top cover wafer substrate. The top cover wafer substrate may cover one side of the main wafer substrate and define a cavity between one side of the main wafer substrate and a hollow space defined by the slotted top cover wafer substrate.

[0164] Preferably, the packaging device includes a slotted bottom cover wafer substrate. The bottom cover wafer substrate may cover the other side of the main wafer substrate and define a cavity between the other side of the main wafer substrate and the hollow space defined by the slotted bottom cover wafer substrate.

[0165] The packaging device (particularly at least one wafer substrate, particularly the at least one grooved wafer substrate) may include one or more holes. These holes may be configured to allow aqueous solutions (particularly bodily fluids) to enter a chamber covered by the at least one wafer substrate. A wafer substrate suitable for forming such a porous surface is CoralPor® porous glass, commercially available from Schott AG, Germany. The hole geometry may be cylindrical, with a hole height preferably 200 to 800 μm, preferably 300 to 600 μm, preferably 360 to 580 μm, a hole diameter preferably 10 to 500 μm, preferably 10 to 300 μm, a preferred hole size of 20 to 150 μm, preferably 32-80 μm, and a preferred porosity of 40-80%, preferably 60-70%, preferably 60-75%.

[0166] Preferably, the encapsulation device completely encapsulates the electronic control device, or the encapsulation device completely encapsulates the electronic control device and the communication device, or the encapsulation device completely encapsulates the electronic control device, the communication device and the biosensor device, wherein the complete encapsulation ensures that the completely encapsulated electronic control device and / or communication device do not come into contact with bodily fluids when the implantable device is implanted in a human or animal, and / or, in particular, ensures that the biosensor device can come into contact with bodily fluids when the implantable device is implanted in a human or animal.

[0167] Typically, the encapsulation device may be composed partially or entirely of at least one non-glass material (e.g., a polymer). In another example, a biosensor device can be encapsulated by embedding it in a sealing compound, which may be biocompatible or may include a biocompatible coating.

[0168] The electronic control device preferably includes one or more of the following components: a microcontroller, particularly a low-power or ultra-low-power microcontroller; one or more integrated circuits (ICs), particularly an ASIC (Application-Specific Integrated Circuit); one or more electronic amplifiers for amplifying electrical measurement signals, particularly current, received from the biosensor device; a light source, preferably an LED, for visually transmitting information to a user of the implantable device, or another output device for transmitting signals, such as acoustic signals, to the user. The electronic control device may be part of the biosensor device or may be connected to the biosensor device.

[0169] An ASIC is a type of integrated circuit (IC) designed and manufactured to perform a specific set of functions in a particular application or system. Unlike general-purpose ICs designed to perform multiple functions, ASICs are customized and optimized for specific applications, offering high performance, lower power consumption, and typically lower cost compared to other solutions. However, the integrated circuit can also be a general-purpose IC programmed to, for example, implement a specific set of functions.

[0170] This particular set of functions may include: controlling one or more electrodes of a biosensor device to perform a measurement process, particularly by measuring the current or impedance between at least two electrodes of the biosensor device when a potential change is applied to at least two electrodes. The measurement is preferably DPV. This particular set of functions may include: receiving measurement signals from the biosensor device and deriving measurement data therefrom. This particular set of functions may include: transmitting measurement data to an external data processing device (e.g., a mobile phone, tablet PC, PC) via a communication device. This particular set of functions may include: receiving electrical energy from radio waves arriving at the implantable device from the outside via a communication device (particularly an RFID device, particularly an NFC device), and preferably using said electrical energy to power the measurement process.

[0171] Preferably, the electronic control device is programmed to implement the specific set of functions.

[0172] DPV stands for Differential Pulse Voltammetry, an electrochemical technique used for signal detection and quantification in a variety of applications. It involves applying a series of voltage pulses to an electrochemical cell and measuring the resulting current response. A preferred electrochemical cell setup for DPV is as follows: a working electrode, a reference electrode, and a counter electrode are placed within the cell. The DPV technique applies a series of voltage pulses to the working electrode. Each pulse typically consists of a step potential, a holding potential, and a pulse width. The step potential provides the driving force for the electrochemical reaction, and the holding potential allows the system time to reach equilibrium before the next pulse. Current measurement: After each voltage pulse, the resulting current response is measured using a potentiostat. This current response corresponds to the electrochemical reaction occurring at the working electrode. The obtained current response is typically plotted against the applied voltage pulses. The resulting DPV curve shows a characteristic peak shape that can provide information about analyte concentration, redox reactions, and other electrochemical processes occurring in the system with the target analyte, thereby facilitating the electrochemical reaction.

[0173] The electronic control device may include at least one program code for implementing a specific set of functions, and the program code may also contain one or more of the following functions; the electronic control device is preferably programmed to implement one or more of the following functions:

[0174] • Process measurement data obtained from the biosensor device, the data source of which is the measurement process performed by the biosensor device;

[0175] • Use encryption algorithms, especially lightweight encryption algorithms, to encrypt measurement data;

[0176] • Establish a data connection between the at least one biosensor device, or correspondingly, the communication device of the at least one implantable device and the communication device of the external computer device, for example by using a data communication protocol, such as a known communication protocol (e.g., TCP / IP), or a custom communication protocol;

[0177] • Data is exchanged between the at least one biosensor device, or correspondingly, the communication device of the at least one implantable device and the communication device of an external computer device, for example by using a data communication protocol, such as a known communication protocol (e.g., TCP / IP), or a custom communication protocol;

[0178] • Data received from the at least one biosensor device, or correspondingly, the communication device of the at least one implantable device, is stored in a data memory, which may be part of or connected to the biosensor device, or correspondingly, the implantable device.

[0179] The communication device is preferably a programmable device, and in particular, is preferably programmed to receive measurement data from an electronic control device in encrypted form and to transmit the measurement data to an external computer device via a wireless connection (in particular via RFID, preferably via NFC or WLAN).

[0180] RFID stands for Radio Frequency Identification. An RFID device is a system or apparatus that uses radio waves to wirelessly exchange data using radio frequency electromagnetic fields. RFID systems for data exchange typically consist of two main components: an RFID transponder and an RFID reader, which can be part of an external computer device, particularly a mobile phone. An RFID transponder is a small electronic device containing a microchip and an antenna. The microchip stores measurement information / data generated by a biosensor device. The antenna enables communication with the RFID reader using radio waves. RFID transponders are preferably passive, but active is also preferable. Passive RFID transponders have no internal power source; instead, they rely on energy emitted by the RFID reader to power themselves. When the reader emits radio waves, the transponder's antenna captures the energy and uses it to activate the microchip, preferably providing power for the measurement process, and transmitting the stored measurement data back to the reader. Active RFID transponders have their own internal power source (usually a battery), which can be part of an implantable device. This allows them to transmit signals continuously or on demand, providing longer reading distances and the ability to store more data. Active transponders are typically used in applications requiring long-distance or real-time tracking. An RFID reader is a device that transmits radio waves and receives signals from RFID transponders. It consists of an antenna, a transceiver for transmitting and receiving radio signals, and a decoder for interpreting data received from the tags. The reader communicates wirelessly with the tags and can read multiple tags within its range simultaneously. When an RFID transponder enters the range of an RFID reader, the reader's radio waves activate the tag. The transponder responds by sending its stored data back to the reader, which then decodes and processes the information. The reader can be connected to a computer or backend system for further processing and utilization of the data collected from the transponders.

[0181] NFC stands for Near Field Communication. NFC devices are devices equipped with NFC technology, enabling them to communicate wirelessly with other NFC-enabled devices or transponders at close range. NFC is a short-range wireless communication technology that operates at a frequency of 13.56 MHz. NFC devices typically include smartphones, tablets, smartwatches, and other electronic devices with built-in NFC chips. These devices can interact with each other by bringing them close together or tapping them.

[0182] NFC enables contactless communication between devices. Simply place two NFC-enabled devices together or tap them, and they can establish a connection and exchange data. NFC allows the transfer of various types of data between devices, such as measurement data. This data exchange can be initiated by the user or triggered by a specific application or service. NFC technology provides a convenient and secure method for short-range wireless communication and data transfer.

[0183] The communication device is preferably an RFID device, and more preferably an NFC device.

[0184] The biosensor device includes at least one sensing element, particularly at least one electrode, which may support a biosensor membrane, such as a membrane containing an aptamer, for measuring the binding of a biomarker (preferably a hormone) to the aptamer during measurement, or correspondingly, the surface concentration of the biomarker on the electrode. The biosensor membrane is configured to contact a bodily fluid for measuring biomass within the fluid. Preferably, the electrode includes a reference electrode and one or more working electrodes, one of which supports the biosensor membrane. The biosensor device may be a stand-alone device. Preferably, the biosensor device may include electronic control devices and / or communication devices. Except for the encapsulation device, the biosensor device can be understood to include the same components as an implantable device. Without an encapsulation device, the biosensor device can be used as an in vitro solution or in vitro in humans or animals, for example, in contact with the skin tissue of the body.

[0185] The biosensor device may be a passive device powered by energy received via a communication device from an external computer device for interrogating the biosensor device. The external computer device (e.g., a mobile phone) may include a communication device capable of transmitting the radio waves to interrogate the biosensor device. The communication device is typically capable of wireless data exchange with at least one biosensor device. In particular, the biosensor device and / or implantable device is a battery-free device / device.

[0186] However, it is also preferred that the biosensor device includes a battery for providing electrical power for operating the measurement process using the biosensor device, and / or for providing electrical power for exchanging data with at least one external computer device. In this context, "external" means that the computer device is separate from both the biosensor device and the implantable device. When the implantable device is implanted into a human or animal, the implantable device is separated from the external computer device.

[0187] The at least one sensing element may be an electrode. In this case, an electrical measurement method (particularly voltammetry or capacitance measurement) is used to measure the at least one biomass.

[0188] The at least one sensing element may be an optical sensing element, which may include a radiation source (especially a light source) and / or a radiation sensor (e.g., a photodetector). In this case, the at least one biomass is measured using optical measurement methods, particularly measurements sensitive to changes in the refractive index of the test material, or measurements sensitive to changes in the amplitude and / or radiation spectrum (spectrum) of the sensing radiation (light) emitted by the radiation source. The optical sensing element may be a photochemical sensing element capable of detecting the presence or concentration change of at least one chemical or biological compound attached to the measurement surface of the photochemical sensing element, the chemical or biological compound arising from contact between the measurement surface and the liquid (especially bodily fluid) to be measured.

[0189] The at least one electrode can be made of gold. It can additionally support at least one or more layers of metal nanoparticles, particularly commercially available gold nanoparticles. Therefore, the measuring surface area of ​​the biosensor membrane is increased – the surface can then be non-planar, but rather surrounding the nanoparticles. A platinum nanoparticle layer can be provided on top of the gold nanoparticle layer. The inventors have unexpectedly discovered that the combined layer of gold and platinum nanoparticles effectively increases the lifetime of the biosensor device, thereby increasing the lifetime of implantable devices. The gold nanoparticles are preferably attached to one or more electrodes, particularly the working electrode. The platinum nanoparticles are preferably attached to the gold nanoparticle layer.

[0190] The present invention also relates to a system for sensing at least one biomass in an animal or human body, the system comprising:

[0191] • At least one biosensor device according to the invention, or correspondingly, at least one implantable device according to the invention.

[0192] • At least one external computer device, which is separate from the at least one biosensor device, or correspondingly the at least one implantable device, the at least one external computer device including a communication device, particularly an RFID, NFC or WLAN device, for wireless exchange of communication device data (particularly measurement data) with the at least one biosensor device, or correspondingly the at least one implantable device.

[0193] The system may further include program code executable by the external computer device, the program code being configured to implement data exchange between the communication device of the at least one biosensor device, or correspondingly, the communication device of the at least one implantable device, and the communication device of the external computer device. In particular, the external computer device may be programmed (especially by using the program code) to implement one or more of the following functions:

[0194] • Establish a data connection between the at least one biosensor device, or correspondingly, the communication device of the at least one implantable device and the communication device of the external computer device, for example by using a data communication protocol, such as a known communication protocol (e.g., TCP / IP), or a custom communication protocol;

[0195] • Data is exchanged between the at least one biosensor device, or correspondingly, the communication device of the at least one implantable device and the communication device of the external computer device, for example by using a data communication protocol, such as a known communication protocol (e.g., TCP / IP), or a custom communication protocol;

[0196] • Decrypt encrypted data, particularly encrypted measurement data received from the at least one biosensor device, or correspondingly, the communication device of the at least one implantable device;

[0197] • Process the data, particularly decrypted data, and especially evaluate the data to obtain measurement values ​​from the data;

[0198] • Data received from the at least one biosensor device, or correspondingly, the communication device of the at least one implantable device, is stored in a data memory, which may be part of or connected to the external computer device.

[0199] It is worth noting that the method for fabricating gold electrodes on a glass wafer substrate is an invention independent of other inventive aspects of the implantable device and the biosensor device. The method includes the following steps:

[0200] - Provide glass wafer substrates;

[0201] - Deposit an electrode structure made of an adhesion-promoting layer to improve gold adhesion on the glass wafer substrate. The adhesion-promoting layer is preferably made of NiCr or chromium, particularly by vapor deposition using a NiCr or Ni source, and especially by performing a photolithography process.

[0202] - Deposit gold on the electrode structure, particularly using vapor deposition with gold atoms.

[0203] Preferably, the method for fabricating gold electrodes on a glass wafer includes any of the following steps:

[0204] • Remove any contaminants from the glass wafer substrate.

[0205] • Use a suitable photoresist to create electrode patterns on the glass wafer substrate.

[0206] • Create a photomask with the desired electrode pattern. This mask will be used to expose the photoresist and create the desired pattern.

[0207] • Prepare the developer solution according to the manufacturer's instructions.

[0208] • Use a suitable NiCr or Ni or gold source for vapor deposition, such as a (gold) evaporator.

[0209] • The glass wafer substrate is immersed in a cleaning solution to remove surface contaminants.

[0210] • Rinse the glass wafer substrate thoroughly with distilled water and allow it to dry.

[0211] • Apply photoresist uniformly to the surface of the glass wafer substrate.

[0212] • Use a spin coater or other suitable method to ensure uniform coverage and a smooth surface.

[0213] • Dry the photoresist according to the manufacturer's instructions.

[0214] • Place the photomask on the glass wafer substrate.

[0215] • Expose the glass wafer substrate to UV light according to the manufacturer's instructions for the photoresist.

[0216] • The exposure will cause the photoresist to become soluble in the exposed area, while remaining insoluble in the mask area.

[0217] • Immerse the exposed coverslip in the developer solution according to the manufacturer's instructions.

[0218] • The developer will remove the soluble photoresist, thereby exposing the electrode pattern.

[0219] • Place the prepared glass wafer substrate with exposed electrode patterns in a vacuum chamber together with a NiCr or Ni or gold source.

[0220] • A vacuum is created in the chamber, and gold is evaporated from a NiCr or Ni or gold source.

[0221] • NiCr or Ni or gold atoms will condense onto the surface of the glass wafer substrate to form a thin NiCr or Ni or gold layer covering the electrode pattern.

[0222] • Depending on specific requirements, post-processing steps, such as annealing or surface finishing, may be required to enhance the adhesion and performance of NiCr, Ni, or gold electrodes.

[0223] By following these steps, NiCr or Ni or gold electrodes can be applied to the glass wafer substrate by photolithography using vapor deposition of gold atoms.

[0224] Each electrode features a complex layered structure to ensure sufficient electrical contact with the biosensor. Each layer contributes to the performance of a single-atom layer bound to the biosensor to generate an electrical signal. Nanolayers are deposited on a wafer to maximize electrical contact with the aforementioned biosensor. This process is performed using atomic layer deposition methods to achieve crucial contact with the adhesion layer.

[0225] In a preferred manufacturing step, a mask / etching preparation is performed on the back / outer side of the wafer: a biocompatible glass wafer with a cut diameter of approximately 100 mm (up to 300 mm) is prepared and cleaned in a preparation solution to improve the adhesion of the first deposited layer.

[0226] During the process steps, a wafer is prepared using a positive mask and a negative mask to allow for the application of a thin-film metallization layer. Advantageously, a glass adhesive layer is used for bonding and provides adequate tensile strength (50 N) to allow for the metallization of the additional layer on the prepared surface.

[0227] The wafer is now ready for additional metallization, either by using electroplating to increase the layer height to 50,000 nm, or by activating the Au surface to allow the biosensor to attach to the electrode.

[0228] Biosensor devices can be fabricated in three stages.

[0229] 1) Activation of the gold layer deposited on the glass is accomplished by connecting the working electrode (gold) to a potentiostat and performing continuous cyclic voltammetry (CV) in 50 mM sulfuric acid. Upon completion of CV, the electrode is cleaned with ethanol and DI water.

[0230] 2) Deposit gold nanoparticles onto the gold layer on top of the glass. This stage is performed to increase the surface area of ​​the gold surface. And finally:

[0231] 3) Activate the biosensor membrane, which includes a self-assembled monolayer (SAM) of the aptamer, which is responsible for permanently attaching the aptamer to the gold platform.

[0232] The biosensor membrane may contain molecules for binding biomarkers from body fluids. These molecules may be aptamers, or correspondingly, antibodies. The hormone is preferably any one of estradiol, LH, or progesterone.

[0233] The present invention is also defined by a method for operating an implantable device or biosensor device according to the invention, the method comprising the following steps:

[0234] a. Contact the biosensor membrane with body fluids;

[0235] b. To bind biomarkers (especially hormones) to biosensor membranes;

[0236] c. Detecting the binding of hormones to a biosensor membrane by electrically controlling at least one metal electrode surface.

[0237] This invention particularly relates to an implantable device for implantation (especially subcutaneous implantation) into an animal or human body, comprising a biosensor device for sensing biomass in vivo (especially the biosensor device according to claim), and an electronic control device for controlling said biosensor device. The invention relates to the following corresponding preferred aspects of said implantable device, wherein the implantable device according to each aspect is claimable by patent claims, or wherein the implantable device according to a combination of multiple aspects is claimable by patent claims:

[0238] According to aspect 1, the implantable device includes

[0239] A packaging device, which is described herein in many respects, is particularly used for at least partially encapsulating at least one of the biosensor device and the electronic control device, wherein the packaging device is at least partially made of glass.

[0240] According to aspect 2, the implantable device, specifically configured according to aspect 1, further includes a communication device, particularly an NFC device, which is connected to an electronic control device for wireless data communication with an external device.

[0241] According to aspect 3, the implantable device, specifically configured according to aspect 1 or aspect 2, is also defined as

[0242] The encapsulation device completely encapsulates the electronic control device, or

[0243] The encapsulation device completely encapsulates the electronic control device and the communication device, or

[0244] The encapsulation device completely encapsulates the electronic control device, the communication device, and the biosensor device.

[0245] The complete encapsulation ensures that, when the implantable device is implanted in a human or animal body, the fully encapsulated electronic control device and / or communication device do not come into contact with bodily fluids, and / or in particular, when the implantable device is implanted in a human or animal body, the biosensor device may come into contact with bodily fluids.

[0246] According to aspect 4, the implantable device, which is configured particularly according to aspect 1 and preferably according to any one of aspects 2 to 3, is also defined as wherein the packaging device comprises at least one wafer substrate, the wafer substrate being substantially made of glass.

[0247] According to aspect 5, the implantable device, which is configured particularly according to aspect 4 and preferably according to any one of aspects 2 to 3, is also defined as

[0248] The packaging device includes at least one wafer substrate, the wafer substrate carrying one or more electrodes on one side of the wafer substrate, and preferably carrying at least one electronic component, particularly the electronic control device, on the other side of the wafer substrate, wherein preferably, the one or more electrodes are connected to the at least one electronic component.

[0249] The present invention particularly relates to a method for operating an implantable device according to any one of aspects 1 to 5, or correspondingly, its biosensor device, the method comprising the following steps:

[0250] a. Contact the biosensor membrane of the biosensor device with body fluid;

[0251] b. To bind a biomarker, preferably a hormone, to the biosensor membrane;

[0252] c. Detecting the binding or surface concentration of hormones on the biosensor membrane by electrically controlling the surface of at least one metal electrode.

[0253] According to aspect 6, the present invention relates to a system for sensing at least one biomass in an animal or human body, the system comprising:

[0254] • At least one biosensor device according to the invention, or correspondingly, at least one implantable device according to any one of aspects 1 to 5, or any other embodiment described herein;

[0255] • At least one external computer device, which is separate from the at least one biosensor device, or correspondingly the at least one implantable device, the at least one external computer device including a communication device, particularly an RFID, NFC or WLAN device, for wirelessly exchanging data, particularly measurement data, with the communication device of the at least one biosensor device, or correspondingly the at least one implantable device.

[0256] According to aspect 7, the present invention relates to program code executed by an external computer device, the program code being configured to implement the at least one biosensor device, or correspondingly, data exchange between a communication device of the at least one implantable device and a communication device of the external computer device. In particular, the external computer device can be programmed to implement one or more of the following functions using the program code:

[0257] • Establish a data connection between the at least one biosensor device, or correspondingly, the communication device of the at least one implantable device and the communication device of the external computer device, for example by using a data communication protocol, such as a known communication protocol (e.g., TCP / IP), or a custom communication protocol;

[0258] • Data is exchanged between the at least one biosensor device, or correspondingly, the communication device of the at least one implantable device and the communication device of the external computer device, for example by using a data communication protocol, such as a known communication protocol (e.g., TCP / IP), or a custom communication protocol;

[0259] • Decrypt encrypted data, particularly encrypted measurement data received from the communication device of the at least one biosensor device, or correspondingly, the communication device of the at least one implantable device;

[0260] • Process the data, particularly decrypted data, and especially evaluate the data to obtain measurement values ​​from the data;

[0261] • Data received from the at least one biosensor device, or correspondingly, the communication device of the at least one implantable device, is stored in a data memory, which may be part of or connected to the external computer device.

[0262] According to aspect 8, the present invention relates to a method for manufacturing a biosensor device and / or implantable device according to the present invention, comprising the following steps:

[0263] • At least one metal electrode surface, particularly an electrode, is prepared on at least one surface of at least one wafer substrate, wherein the wafer substrate is preferably made of or contains gold, and the wafer substrate is preferably a glass wafer substrate;

[0264] • Prepare the biosensor membrane on the surface of the at least one metal electrode;

[0265] • Optionally: Electronic control devices are arranged on the at least one wafer substrate;

[0266] • Optionally: Use the at least one wafer substrate to fabricate a packaging device for encapsulating the biosensor device, or correspondingly, the chamber of the implantable device.

[0267] According to aspect 8, the method described according to aspect 8 includes the following steps:

[0268] • Provide multiple wafers (two or more) containing multiple wafer substrates for parallel fabrication of multiple implantable devices, wherein the multiple wafers are preferably arranged in a stacked manner;

[0269] • Welding the plurality of wafer substrates, particularly at splice points or regions, preferably to create at least one chamber, such that the resulting stack of welded glass wafer substrates, particularly the at least one chamber, encapsulates the biosensor device, or correspondingly at least one component of the implantable device: electrodes, electronic control devices, particularly microcontrollers, one or more amplifiers, RFID (preferably NFC) chipsets; antennas.

[0270] • Optionally: Cut at least one or more stacked and soldered wafer substrates from the stacked and soldered wafers, the wafer substrates specifically carrying and / or encapsulating one or more of the components.

[0271] Currently available implantable sensor devices rely on batteries to power their operation for a limited time, after which the batteries need to be replaced or wirelessly recharged. In either case, there are several disadvantages to the patient's health and well-being. Removing the battery requires additional implantation surgery, which involves all the associated health risks and potential surgical site infections. Recharging the battery takes time, which inconveniently restricts the patient's movement or body posture during wireless power transmission.

[0272] Therefore, according to this particular aspect of the invention, the object of the invention is to provide a biosensor device, or correspondingly, an implantable device, which avoids the aforementioned disadvantages, such as long charging time or additional implantation surgery.

[0273] The problem is solved by the biosensor device according to implementation 1 and the method according to implementation 15. Furthermore, other embodiments are described by implementations 2 through 14 and 16, and other preferred configurations are described throughout the patent application. As disclosed herein, any of the mentioned implementations of the biosensor device can be used alone or in combination with one or more other implementations and with any aspect of the invention.

[0274] According to embodiment 1 of the invention, the biosensor device is configured to be implanted in a human or animal body, particularly subcutaneously. Such an implantable biosensor device is also referred to as an "implantable device" and may include an encapsulation device for at least partially encapsulating at least one of the biosensor device and an electronic control device. According to embodiment 1, the biosensor device (1i) or implantable device (1i) includes at least one metal electrode surface, preferably a portion of an electrode, particularly an electrochemical sensing electrode, for detecting voltage equivalent data for quantifying biomass, and

[0275] Electronic control device (2i),

[0276] The electronic control device (2i) includes

[0277] • At least one amplifier (3i), and

[0278] • A microcontroller (5i) for digitizing voltage equivalent data into digital measurement data, and

[0279] • A communication device (6i), particularly an NFC device (6ii), wherein the NFC device (6ii) is configured to transmit digital measurement data of the electrochemical sensor to an external device (28i) (uplink) and receive specific data (100i) from the external device (128) (downlink), and is also configured to be powered by an external computer device (108) so that the biosensor device (101) is fully operational.

[0280] Preferably, the microcontroller (5i) is programmed to encrypt the digitized data into encrypted data, and wherein the encrypted data is sent to the (near-field) communication device (6i), and wherein the encrypted data is further transmitted to an external computer device (8i) via a near-field antenna (7i).

[0281] Preferably, the biosensor device includes an electrochemical sensor for sensing at least one biological / biochemical analyte (biomass) and electronic circuitry for monitoring body hormone levels, particularly hormone levels associated with a woman's reproductive cycle or during in-vitro fertilization, wherein the electronic circuitry includes at least one amplifier (4i) for converting and amplifying a current derived from the analyte into voltage equivalent data relative to a reference voltage level; and at least one amplifier for generating the reference voltage level; and, preferably, an ultra-low power microcontroller (5i) for digitizing the voltage equivalent data into digital data; and, preferably, includes a (near-field) antenna. The near-field communication (NFC) device is configured to transmit digital data of an electrochemical sensor to an external computer device (8i; uplink) and receive specific data from the external computer device (downlink), and is further preferably configured to be powered by the external computer device so that the biosensor device is fully operational, wherein the microcontroller (5i) is preferably programmed to encrypt the digital data into encrypted data, wherein the encrypted data is sent to the communication device, and wherein the encrypted data is further transmitted to the external computer device via a (near-field) antenna (7i).

[0282] In this context, an electrochemical sensor is understood as a device that measures and detects chemical substances by converting chemical reactions occurring on an electrode into electrical signals. It utilizes electrochemical principles to sense and quantify multiple analytes or target substances in a sample (i.e., at an implantation site in a human or animal). The biosensor device according to the invention preferably generally comprises an electrochemical sensor, which includes at least one electrode, or correspondingly, at least one metal electrode surface.

[0283] To measure analyte concentration (also known as analyte measurement or analyte sensing), the sensor preferably includes a reference electrode. The reference electrode provides a stable potential against which the analyte reaction is compared, thereby ensuring accurate measurement. By monitoring changes in current or potential generated by the analyte reaction, the sensor can determine, and in particular, the concentration of the target substance in the sample.

[0284] In the context of this article, biomass or bio / biochemical analytes are understood to be, for example, hormones in the human or animal body, particularly hormone levels associated with the female reproductive cycle or during in-vitro fertilization.

[0285] In the context of this paper, the amplifier used to convert and amplify the current derived from the analyte into voltage equivalent data is understood as a current-to-voltage converter, such as a transimpedance amplifier, which is implemented almost entirely using one or more operational amplifiers. The voltage equivalent data corresponds to the voltage equivalent level of the data generated through a resistor.

[0286] The electrochemical sensor is connected to an electronic circuit via a working electrode. Therefore, the electronic circuit includes at least one working electrode for electrically connecting the electrochemical sensor to the electronic circuit, particularly for connecting it to the amplifier, especially the transimpedance amplifier, for converting the current provided by the electrochemical sensor into a voltage, i.e., into a voltage equivalent value, also known as voltage equivalent data, based on the current data that serves as data from the electrochemical sensor.

[0287] In the context of this paper, the reference voltage corresponds to the voltage reference level generated by an operational amplifier combined with a voltage divider circuit topology, and subsequently buffered by the amplifier.

[0288] In the context of this paper, ultra-low power microcontrollers are understood as microcontrollers that enable data to be processed with the minimum amount of system power required.

[0289] In the context of this article, Near Field Communication (NFC) devices are understood as NFC chip tags, also known as NFC tags or simply NFC chips, which are small electronic devices containing an integrated circuit (IC) with an NFC antenna (i.e., a near-field antenna). The tag is designed to wirelessly store and transmit data using NFC technology. NFC chip tags are understood as passive devices, meaning they do not have their own power source. Instead, they rely on power provided by, for example, an external device. When an external device is very close to the NFC tag, particularly within a few centimeters, the external device generates an electromagnetic field that induces a small current in the tag's near-field antenna. This current powers the NFC chip, enabling it to communicate with the external device.

[0290] The NFC chip tag can be embedded or attached to a printed circuit board and / or a flexible circuit board. The data stored in the NFC tag can vary from simple text or URLs to more complex information, such as biosensor device details, product information, or commands for specific operations.

[0291] The NFC chip tag is versatile and can be programmed and reprogrammed multiple times, making it highly flexible in its applications. It can be read and written by NFC-enabled devices such as smartphones, tablets, or dedicated NFC readers. By tapping an NFC-enabled device (e.g., an external device to a mobile device) or bringing it close to the NFC tag, a user can read information stored in the tag or initiate specific operations based on the tag's programming.

[0292] The biosensor device is fully operational when it is able to convert current from an electrochemical sensor to an operational amplifier into a corresponding voltage and pass it to an ultra-low power microcontroller for digitization and encryption, send it to an NFC device, and transfer it from the NFC device to an external device that decrypts the transmitted encrypted data. The biosensor device is not fully operational when it is merely powered by an external device at NFC frequencies without authorization (e.g., authorization to decrypt).

[0293] The ultra-low power microcontroller is programmed to encrypt digitized data into encrypted data, and the encrypted data is sent to a near-field communication device, and the encrypted data is further transmitted to an external device.

[0294] To send encrypted data from the microcontroller to the NFC device, an Integrated Circuit Bus (I2C) interface is used. The I2C interface is a widely used serial communication protocol that allows multiple devices to communicate with each other via a shared bus. Although I2C and NFC are different protocols, they can be used in combination. For example, an NFC-enabled device can be connected to a microcontroller or other peripherals using the I2C interface. This allows the NFC device to communicate with the microcontroller, which can then control other components or perform additional processing based on the NFC data. In such a setup, the NFC device handles NFC-specific communication, while the I2C interface facilitates communication between the NFC device and the microcontroller. In this case, the microcontroller also includes an encryption algorithm and is therefore programmed to encrypt the provided digitized data. The encrypted data is then sent to the NFC via the I2C interface. This feature has the effect that the ultra-low power microcontroller will not start any operation until it receives a valid command sequence from an authorized external device (e.g., an NFC-enabled mobile phone running the corresponding application), while the power required to generate the voltage level (i.e., the acquired voltage level) needed to run the biosensor device can be generated by any NFC-enabled external device or any external device with the same radio frequency (RF) as the NFC frequency without authorization.

[0295] While the I2C communication protocol specifications for baud rate and byte format are generic, the data packets (i.e., byte streams) exchanged between the ultra-low power microcontroller and the NFC chip tag are customized. This means that some transmission bytes, for example, that the microcontroller needs to perform, are encoded, such as for acquiring voltage equivalent data and data transfer to the NFC, or for control parameters used to operate the biosensor device itself. Preferably, the byte stream or data packet generated for NFC transmission includes a byte field occupied by a unique ID number for the implantable device, plus encrypted data obtained from the biosensor / temperature channel, control parameters, and a security password for data decoding, resulting in a total data payload of 1kB in each NFC transmission (downlink / uplink).

[0296] Because the biosensor device is preferably battery-free, it is not affected by the aforementioned drawbacks, thus remaining inside the body (i.e., subcutaneously), its duration is not limited by the stored energy level, and only requires the presence of an NFC-enabled mobile phone in the vicinity for each cycle of device operation. From the patient's perspective, this is not too inconvenient, as readings can be collected intermittently rather than continuously for the device's intended application (i.e., monitoring of body hormones), thereby reducing the impact of the reading procedure on the patient's normal daily life. Furthermore, the implantable electronic circuitry described herein does not actively stimulate subcutaneous body tissue through any physical conversion mechanism (which could be electrical, optical, or ultrasonic) like other commercial devices, making its use in vivo safer and more likely to be approved by regulatory agencies worldwide.

[0297] In preferred implementation 2, the external computer device is an NFC-enabled portable PC, tablet PC, or mobile phone, and is programmed to decrypt the transmitted encrypted data. This feature ensures that no other mobile device is authorized to interact with the biosensor device. Furthermore, this makes data reading particularly easy for patients, who often carry their mobile devices with them, thus eliminating the need for additional devices. Additionally, for example, the data can be stored on the mobile device and easily sent to a doctor for evaluation.

[0298] In a preferred embodiment 3 according to the invention, the decrypted data is visualized on the display of an external computer device, particularly a touchscreen.

[0299] Visualization of transmitted data allows patients to quickly obtain an overview of relevant hormone concentrations, for example, without the need for additional technical equipment.

[0300] In a preferred embodiment 4 according to the invention, the external device is located at a distance of less than 2 cm or 2 cm from the biosensor device for data transmission, i.e., establishing an uplink or downlink. Such a distance is particularly suitable for subcutaneously implanted biosensor devices. The uplink / downlink and power supply lines can operate within a maximum gap distance of 2 cm between the implantable device and the external communication device (especially an NFC source) without affecting the electronic performance specifications (i.e., nominal acquisition voltage level and transmission quality) set for the implantable device.

[0301] In a preferred embodiment 5 according to the invention, the digitization sampling rate comprises a rate of 24 samples per second and a resolution of 10 bits, or a rate of 24 samples per second and a resolution of 10 bits. Such values ​​are useful because, for data collected from biosensors / temperature, the resolution associated with the number of bits will set the accuracy limit for the implantable device, while the number of samples per second will affect the size of the data packet (payload) for each NFC transmission, which is preferably set to a limit of 1 kB.

[0302] In a preferred embodiment 6 according to the invention, the digitized data is temporarily stored in the microcontroller. For data encoding, the data and intermediate results derived from the logical encryption operation (XOR) are temporarily stored in the microcontroller's limited RAM memory before the processed encrypted data is included in the NFC data packet for transmission.

[0303] In a preferred embodiment 7 according to the invention, the reference voltage level is generated during sensing of at least one biochemical analyte. This feature enables higher measurement accuracy because it allows for taking into account variations in the reference voltage level that may occur for any reason. By fine-tuning the reference voltage level, different chemical processes more relevant to the target analyte (i.e., the selection of specific chemical functional groups and / or the improved geometric alignment of specific molecular structures) are initiated, thereby obtaining better accuracy of the measured signal, and this does not require increasing the resolution (number of bits) of the digital electronic equipment.

[0304] In a preferred embodiment 8 according to the invention, the electronic circuit includes at least three working electrodes configured to sense at least three analytes, particularly different analytes, for the electrochemical sensor. In other preferred embodiments of the invention, the electronic sensor includes two, four, five, or even more working electrodes configured to sense one, two, three, four, five, or even more analytes. This advantageously allows for the measurement of the same analyte using multiple working electrodes, thereby increasing the accuracy of the biosensor device.

[0305] In a preferred embodiment 9 according to the invention, the sensing of the chemical analytes is performed simultaneously, such that the current generated simultaneously by the electrochemical sensor is processed in parallel by electronic circuitry. This feature advantageously enables the simultaneous monitoring of the concentration of each different analyte, for example, by visualizing them on a mobile phone display.

[0306] In a preferred embodiment 10 according to the invention, the digitized data also includes the temperature of the biosensor device. Information about the temperature level is provided via an uplink communication channel because the microcontroller advantageously includes an internal temperature indicator built into the die, thereby allowing, for example, further or more detailed analysis of the provided patient-specific data.

[0307] In a preferred embodiment 11 according to the invention, the specific data may include user parameters, including data for setting the ID number of the biosensor device, the operating status of the biosensor device, or control data for the sensing process of the biosensor device. This feature also allows for the personalization of the biosensor device, particularly for patient-specific implantable sensors.

[0308] In a preferred embodiment 12 according to the invention, the biosensor device includes an optical indicator, particularly a light-emitting diode (LED), configured to optically indicate the operating status of the biosensor device to the user. For example, when the NFC interface of a mobile phone is successfully powered, a small embedded LED provides visual feedback on the operation performed on the implantable side. This, for example, makes it easier to troubleshoot the biosensor device without having to remove the implanted biosensor device.

[0309] In a preferred embodiment 13 according to the invention, the biosensor device includes a rigid and / or flexible printed circuit board (PCB). Specifically, the flexible board can be used to avoid mechanical stress in the biosensor device structure and / or to achieve a particular advantageous shape for the biosensor device. However, the component can also be mounted directly on a wafer substrate, particularly without using a PCB.

[0310] In a preferred embodiment 14 according to the invention, the (near-field) antenna is wound around a rigid PCB with N turns, wherein N is preferably selected from the range of 15 to 25 turns, more preferably 18 to 22 turns, or N=20, and / or wherein the antenna comprises enameled copper with a thickness of 0.15 mm. The internal NFC antenna allows AC radio frequency waves generated by, for example, a mobile phone to be continuously captured by the NFC antenna, which is wound around the outer boundary of the circuit board and converted by the NFC chip tag itself into an equivalent DC level (referred to as the acquisition voltage level), which can then be used for the rest of the embedded electronics.

[0311] In implementation 15 of the present invention, this problem is also solved by a method of monitoring and analyzing substances (particularly the levels of bodily hormones in the human or animal body, and especially hormone levels related to the female reproductive cycle or in vitro fertilization), the method comprising the following steps:

[0312] - Provides a biosensor device, preferably comprising an electrochemical sensor for sensing at least one chemical analyte; and

[0313] - Provide an electronic control device, particularly an electronic circuit, wherein the electronic control device includes:

[0314] At least one amplifier, specifically configured to convert and amplify the current derived from the analyte into voltage equivalent data relative to a reference voltage level, and

[0315] Preferably: at least one amplifier for generating a reference voltage level, and

[0316] Preferably, an ultra-low power microcontroller is used to digitize the voltage equivalent data into digital data, and

[0317] A communication device, particularly a near-field communication (NFC) device including, for example, a near-field antenna, wherein the communication device is configured to transmit digital data of the biosensor device or electrochemical sensor to an external computer device (uplink) and receive specific data from the external computer device (downlink), and is further preferably configured to be powered by the external computer device, thereby enabling the biosensor device to operate fully.

[0318] Preferably, the microcontroller is programmed to encrypt the digitized data into encrypted data, and the encrypted data is sent to the communication device, such as an NFC device, and the encrypted data is further transmitted to the external computer device.

[0319] - Optionally: connecting the electrochemical sensor to an electronic sensor device, including at least one working electrode; and

[0320] - Preferably: the biosensor device is powered by an external computer device, preferably using wireless connections of these devices, particularly using radio waves; and

[0321] - Optionally, the transmitted and decrypted data can be visualized on the monitor of an external computer device.

[0322] In a preferred implementation 16 of the biosensor device (particularly the electronic control device, especially the electronic circuitry), the acquisition and amplification of ion currents are simultaneously generated by a total of three chemical analytes (or species) contained in a physiological solution and / or liquid, subsequently digitized within an ultra-low power microcontroller, and finally transmitted via a near-field communication (NFC) interface to an external mobile phone located at a distance (preferably less than 2 cm) from the electronic circuitry. The same mobile phone is responsible for supplying the necessary power (energy harvesting) to the implantable electronic device via a radio frequency field (13.56 MHz = standard) generated by NFC. This enables the electronic device to operate at full capacity to perform the aforementioned electrochemical harvesting task, as it does not rely on any form of stored internal energy or battery. The implantable electronic circuitry is intended for monitoring body hormone levels directly related to a woman's reproductive cycle under normal physiological conditions or during in vitro fertilization (IVF) treatment.

[0323] Amperometric measurement is a type of electrochemical analysis that involves measuring the current flowing through a system. Specifically, it involves detecting and quantifying substances based on changes in current generated by the electrochemical reaction of a target analyte or chemical substance at an electrode. In an amperometric measurement, an electrode is immersed in an electrolyte solution containing the analyte. A potential is applied to the electrode, which initiates an electrochemical reaction involving the analyte. As the reaction proceeds, electrons transfer between the electrode and the analyte, producing a measurable current. This current is proportional to the concentration of the analyte in the solution. Attached Figure Description

[0324] Other preferred configurations of the implantable device, biosensor device, and system of the present invention, as well as the method of the present invention, can be derived from the following description of exemplary embodiments in conjunction with the accompanying drawings and description. Unless otherwise stated or clearly indicated by the context, the same components of the exemplary embodiments are characterized by substantially the same reference numerals. Specifically:

[0325] Figure 1 A schematic cross-sectional view of an implantable device according to an embodiment of the present invention is shown.

[0326] Figure 2a It shows Figure 1 A schematic perspective view of an implantable device.

[0327] Figure 2b A top view of the top surface of the wafer substrate carrying the electrode of an implantable device according to an embodiment of the present invention is shown.

[0328] Figure 2c A top view of the top surface of a wafer substrate carrying another electrode in an implantable device according to another embodiment of the present invention is shown.

[0329] Figure 3 A top view is shown of a portion of a circular wafer containing multiple electrode assemblies, from which this portion is cut for fabricating multiple such... Figure 2b The wafer substrates for carrying electrodes shown are each used in an implantable device according to an embodiment of the present invention.

[0330] Figure 4a A perspective side view of an implantable device according to an embodiment of the present invention is shown, the implantable device being made of a top wafer substrate, a middle wafer substrate and a bottom wafer substrate.

[0331] Figure 4b It shows that according to Figure 4a A perspective side view of the intermediate wafer substrate of an implantable device, wherein the intermediate wafer substrate is shown inverted.

[0332] Figure 4c It shows that according to Figure 4a A perspective side view of an implantable device, in which the implantable device is shown upside down.

[0333] Figure 5 The left side shows the basis Figure 4a A perspective view of the intermediate wafer substrate of the implantable device, and a collection of five vertically bound diagrams shown on the right side of the page, from top to bottom: According to Figure 4a A top view of the first (top) wafer substrate of the implantable device, and a cross-sectional view taken along line AA; according to Figure 4a A top view of the second (intermediate) wafer substrate of the implantable device, cut along length L and perpendicularly through... Figure 4a A cross-sectional view of the center of the top surface of the wafer substrate of the implantable device, and a cross-sectional view taken along a line extending from the via 32; according to Figure 4a A bottom view of the second (intermediate) wafer substrate of the implantable device, according to Figure 4a A bottom view of the bottom side of the third (bottom) wafer substrate of the implantable device, and a cross-sectional view taken along line BB.

[0334] Figure 6 A system according to the invention is shown according to an embodiment, which uses an implantable device according to the invention and a biosensor device according to the invention, respectively.

[0335] Figure 7 A schematic diagram of an embodiment of the electronic circuitry of a biosensor device according to an embodiment of the present invention is shown.

[0336] Figure 8 The working principle of a biosensor device according to an embodiment of the present invention is shown.

[0337] Figure 9 The layout of the rigid PCB back side of a biosensor device according to one embodiment of the present invention is schematically shown.

[0338] Figure 10 The layout of the rigid PCB front side of a biosensor device according to one embodiment of the present invention is schematically shown.

[0339] Figure 11 Three graphs are shown showing measurement data obtained using a biosensor device according to an embodiment of the invention.

[0340] Figure 1 A schematic cross-sectional view of an implantable device 1 according to an embodiment of the present invention is shown. The implantable device 1 includes a biosensor device 2 and a packaging device 3. The biosensor device 2 includes all the functional components necessary for the implantable device to function properly, except for the packaging device 3, which is specifically designed to provide long-term operation of the biosensor device within living tissue and to provide biocompatibility to prevent or reduce the risk of rejection or adverse reactions due to various factors, including the body's immune response or the release of harmful substances due to degradation of the implantable device. In this case, the outer layer of the packaging device is made substantially entirely of glass, except for electrodes and conductive traces (not shown) extending on the surface of a top wafer substrate 7a. The top wafer substrate 7a is part of the biosensor device 2 and additionally forms part of the packaging device 3. Specifically, the packaging device is made substantially entirely of glass, and the electrodes are disposed on the top surface 3a of the packaging device 3. The packaging device 3 is made substantially entirely of glass, particularly Borofloat® from Schott AG, Germany. The top glass wafer 7a, the middle glass wafer substrate 7b, and optionally the bottom glass wafer substrate 7c include through-holes 12 filled with a metallic conductive material (e.g., tungsten or NiFe). Such glass wafer substrates can be cut from a larger wafer, which can be commercially available from HermeS® wafers manufactured by Schott AG in Germany. The three glass wafer substrates 7a, 7b, and 7c are connected here by glass welds 8, which hermetically seal the chambers 9a and 9b inside the packaged device.

[0341] Electrodes 4a, 4b, and 4c are sensing elements of the biosensor device 2 and are configured to perform voltammetric measurements, particularly DPV, when an implantable device is implanted into tissue. The electrodes include: a counter electrode 4b, a planar gold electrode, attached to the top surface 3a of the glass wafer substrate 7a; a reference electrode 4c, also a planar gold electrode, attached to the top surface 3a of the glass wafer substrate 7a; and a working electrode 4a, also a planar gold electrode with a functionalized surface attached to the top surface 3a of the glass wafer substrate 7a. The functionalized surface is simplified as a stack of layers 10. However, the actual three-dimensional atomic surface structure (not shown) is more complex. Alternatively, other numbers of electrodes may be provided to allow for electrical measurements; preferably, four electrodes may be provided.

[0342] Furthermore, two or more electrodes can typically be provided on different surfaces of the implantable device, packaging device, or biosensor device, particularly on any wafer substrate. For example, one or more electrodes can be provided on a first (outer) surface 3a of the packaging device, and one or more electrodes can be provided on a second (outer) surface 3b of the packaging device, which may be opposite to the first surface 3a.

[0343] The stack of layers 10 is produced by the layer-by-layer, or correspondingly, stepwise deposition of different nanoparticles or molecules on a previously existing surface. The stack of layers 10 is also referred to as a biosensor membrane 10.

[0344] The biosensor membrane 10 includes a layer 10a of gold nanoparticles deposited on the planar gold surface of electrode 4a. While the gold nanoparticle layer 10a can typically be omitted, it is advantageous for improving the detection signal. The gold nanoparticles have a diameter of approximately 60 nm, while other particle sizes in the range of 10 nm to 200 nm or 30 nm to 100 nm are also preferred. The effect of using a gold layer is that the entire gold surface interacting with the body fluid is generally larger than the planar surface of electrode 4a. Therefore, the surface provided by the nanoparticles and available for binding aptamers to electrode 4a is significantly expanded. Consequently, the number of binding sites for binding biomarkers (hormones or other targets in body fluids) is significantly increased, and signal quality, particularly the signal-to-noise ratio of the measurement, is improved.

[0345] In this configuration, an additional layer 10b of nanoparticles is provided on top of the gold nanoparticle layer 10a. The additional layer 10b of metal nanoparticles (which in this case are platinum nanoparticles) was found to improve the lifetime and reliability of the biosensor membrane 10.

[0346] On top of nanoparticle layers 10a and 10b, aptamer 10c is deposited and bonded to the nanoparticles to obtain biosensor membrane 10. Thiolized aptamers tend to form self-assembled monolayers (SAMs) on the metal surfaces of the nanoparticles, which helps improve the binding stability and permanent adhesion of the aptamers to the gold / platinum platform.

[0347] The biosensor is prepared in three steps:

[0348] 1) Activating the gold layer deposited on the glass is accomplished by connecting the working electrode (gold) to a potentiostat and applying continuous cyclic voltammetry (CV) in an acid solution; this optional step of activating the gold electrode typically includes a cleaning or conditioning process to remove any contaminants or oxide layers, thereby ensuring better electrical contact and performance.

[0349] 2) Deposit gold nanoparticles on a gold layer on top of the glass, and optionally, provide platinum nanoparticles;

[0350] 3) Provide an adaptor layer.

[0351] The electronic control device 11 here includes a printed circuit board 11a, which carries an integrated circuit 11b for controlling electrodes 4a, 4b, 4c and correspondingly for controlling the electrical measurement of at least one biomass. The electronic control device 11, particularly traces (not shown) on the printed circuit board 11a leading to the integrated circuit 11b, is connected via wires 13 to vias 12 leading to the electrodes. Furthermore, the electronic control device 11 here includes a communication device 11c, which in this case is an RFID device, particularly an NFC chipset. The NFC chipset is connected to the integrated circuit 11b via traces (not shown) on the PCB 11a. Furthermore, in this case, the NFC chipset is connected to an antenna 15 via vias 12 on the PCB 11a and wires. The antenna 15 can be mounted anywhere on the biosensor 2 or, correspondingly, on the implantable device 1, preferably inside the package 3. Preferably, the antenna 15 is mounted on a wafer substrate or a portion of the package 3, preferably on the same wafer substrate that also carries the electronic control device 11.

[0352] Electrodes 4a, 4b, and 4c are connected to the electronic control device via through-hole 12. The advantage of using a substrate wafer with through-hole 12 is that the biosensor device 2 can be arranged more compactly, and therefore the implantable device 1 can also be arranged more compactly. Each of electrodes 4a, 4b, and 4c can be connected to at least one through-hole 12 via a gold trace, which can be disposed on a gold surface and extend from the gold electrode to a gold bonding pad (see [reference]). Figure 2b .exist Figure 1 In the middle, electrode 4a is directly installed into the through hole located below electrode 4a, and the same connection path is set for electrodes 4b and 4c.

[0353] The electronic control device 11 is attached here to the wafer substrate, which is the intermediate wafer substrate 7b. However, it is also generally preferred that the electronic control device 11 be attached to the same wafer substrate that also carries at least one electrode, preferably on the side 3c of the wafer substrate opposite to the side 3a carrying the electrode.

[0354] In this case, integrated circuit 11b is an ASIC.

[0355] Electronic control device 11, particularly integrated circuit 11b, is configured, and specifically programmed, to provide electrical measurement for acquiring a measurement signal, or to repeatedly acquire a measurement signal for measuring the concentration of a biomarker in a body fluid or a time-dependent change in a signal in the body fluid, wherein the signal represents the amount of a biomarker bound to biosensor membrane 10. This measurement can be triggered by a schedule or by an external computer device, such as a smartphone with a control application installed, which the user can then operate to initiate the measurement (e.g., by touching the “Start” area in a graphical user interface provided by the control application on the screen of the mobile phone device).

[0356] Preferably, the electrical measurement is a volt-ampere method measurement, particularly a DPV measurement:

[0357] Typically, DPV stands for differential pulse voltammetry, an electrochemical technique used for signal detection and quantification in a variety of applications. It involves applying a series of voltage pulses to an electrode in contact with an electrolyte that forms a bodily fluid (e.g., interstitial fluid) and measuring the resulting current response.

[0358] Electrode setup: A working electrode 4a, a reference electrode 4c, and a counter electrode 4b are provided. Alternatively, other numbers of electrodes may be provided to allow for electrical measurements; preferably, four electrodes may be provided.

[0359] DPV technology applies a series of voltage pulses to the working electrode 4a. These pulses typically consist of a step potential, a holding potential, and a pulse width. The step potential provides the driving force for the electrochemical reaction, while the holding potential allows the system time to reach equilibrium before the next pulse.

[0360] Current measurement: After each voltage pulse, the resulting current response is measured using a potentiostat. This current response corresponds to the electrochemical reaction occurring at the working electrode.

[0361] The obtained current response is typically plotted against the applied voltage pulse. The resulting DPV curve shows a characteristic peak shape, which can provide information about analyte concentration, redox reactions, and other electrochemical processes occurring in the system with the target analyte, thereby promoting electrochemical reactions.

[0362] Figure 2a It shows that according to Figure 1 A schematic perspective view of an implantable device 1, showing the actual dimensions of the device, with a width W = 2.0 mm, a height H = 2.0 mm, and a length L = 16 mm. The implantable device has rounded edges and corners (not shown) and can be implanted by a surgeon using a syringe.

[0363] Figure 2b A top view of the top surface of the wafer substrate 7a' of the implantable device 1 according to an embodiment of the present invention is shown. This embodiment can be used for... Figure 1 In step a, the wafer substrate 7a' has a via 12. Electrodes 4a', 4b', 4c', and 4d' are deposited on the top surface 3a of the wafer substrate. Here, four electrodes are provided, each of which is connected to a bonding pad 14b via a conductive trace 14a. The conductive trace 14a and the bonding pad 14b are deposited on the same surface 3a together with gold electrodes in the same manufacturing step and are made of the same gold.

[0364] Figure 3 A shows a top view of a portion of a circular wafer 40 containing multiple electrode assemblies, from which this portion is cut for fabricating multiple... Figure 2b The wafer substrate 7a' shown carries electrodes, each of which is used in an implantable device according to an embodiment of the invention. The wafer substrate 40 is provided with component vias 12 that perpendicularly penetrate the wafer surface and are filled with a conductive metal, such as tungsten. The wafer 40 is a HermeS® glass wafer from Schott AG, Germany.

[0365] Figure 2c A top view of the top surface of a wafer substrate 7a” carrying another electrode in an implantable device according to another embodiment of the invention is shown. The wafer 40 is a HermeS® glass wafer from Schott AG, Germany. Four electrodes 4a”, 4b”, 4c”, and 4d” are deposited on the top surface 3a” of the wafer substrate 7a”. The four electrodes almost completely cover the surface of the wafer substrate, or correspondingly, a portion of the surface, selected from the range of 80 to 100%, preferably 90 to 100%, preferably 90 to 99%. Using a larger electrode surface results in improved signal quality when performing electrical measurements because it provides a greater number of binding sites for biomarkers.

[0366] Figure 4aAn implantable device 21 according to an embodiment of the present invention is shown in a perspective side view, the implantable device being made of a top wafer substrate 27a, an intermediate wafer substrate 27b, and a bottom wafer substrate 27a. The top wafer substrate 27a and the bottom wafer substrate 27a are configured to have a groove shape and are arranged as covers for covering components located on the top and bottom sides of the intermediate wafer substrate 27b, respectively.

[0367] Figure 4b It shows that according to Figure 4a A perspective side view of the intermediate wafer substrate 27b of the implantable device, wherein the intermediate wafer substrate is shown upside down. On the bottom side of the intermediate wafer substrate 27b, several components of the electronic control device of the biosensor device of the implantable device 21 are attached: an integrated circuit (ASIC) 31b for electrically controlling electrodes 24a, 24b, 24c, 24d, and a communication device 31c that serves as an NFC device and includes an NFC antenna 35.

[0368] and Figure 4b The view of the intermediate wafer substrate 27b is similar. Figure 5 The left side shows the basis Figure 4a A perspective top view of the intermediate wafer substrate 27b of the implantable device shows four gold electrodes 24a, 24b, 24c, and 24d arranged parallel to each other and spaced apart along a length L of the intermediate wafer substrate 27b. The electrodes occupy almost the entire surface of the intermediate wafer substrate 27b. The intermediate wafer substrate 27b is a HermeS® glass wafer from Schott AG, Germany, having through-holes 32 perpendicular to the top surface of the wafer substrate and filled with a conductive metal, such as tungsten. Each electrode contacts the through-hole 32 and is connected through the through-hole to an ASIC 31b, which can be directly deposited on the bottom surface of the intermediate wafer substrate 27b without using a PCB to place electronic components, or using a PCB (not shown) to place the electronic components. According to this embodiment of the invention, the use of a wafer with through-holes allows for the implementation of a compact solution for an implantable device. Those skilled in the art can readily modify the features of the implantable device 21 with other preferred features and configurations as disclosed herein.

[0369] Figure 5 The right side of the page displays a collection of five vertically bound illustrations, from top to bottom:

[0370] i. According to Figure 4aA top view of the first (top) wafer substrate 27a of the implantable device 21, and a cross-sectional view taken along line AA, showing a through-hole or aperture 39. The first (top) wafer substrate 27a serves as a cover plate for covering the surface 23a carrying four gold electrodes 24a, 24b, 24c, and 24d. One or two of the electrodes carry a biosensor membrane, for example, as shown in [reference]. Figure 1 The biosensor membrane 10 is described. Other electrodes are gold electrodes used as reference or counter electrodes for voltammetry (specifically, DVP) measurements. A cover plate 27a protects the biosensor membrane and electrodes from mechanical damage that may occur during implantation or prolonged use within the patient. The porous membrane formed by the main wall of the top wafer substrate and the cover plate 27a allows bodily fluids to easily enter the internal volume encompassed by the top surface of the grooved cover plate 27a and the intermediate wafer substrate 27b, which is welded to the intermediate wafer substrate 27b, thus securing the cover plate firmly and inextricably. The main wall of the cover plate, and possibly its flange surrounding the outer edge of the main wall, is made, for example, of CoralPor® porous glass commercially available from Schott AG, Germany. When the wafer substrates are readily equipped with, for example, electronic components and electrodes plus the biosensor membrane, and precisely aligned with each other, welding is achieved via laser welding, particularly femtosecond laser welding, at the weld lines and weld areas indicated by numeral 28.

[0371] ii. According to Figure 4a A top view of the second (intermediate) wafer substrate 27b of the implantable device; for illustrative purposes, the vias 32 are shown as black dots here, but they are actually not visible because they are covered by gold electrodes.

[0372] iii. Cut along length L and pass perpendicularly through according to Figure 4a A cross-sectional view of the center of the top surface of the wafer substrate of the implantable device, particularly showing a via 32 extending through the entire height of the intermediate wafer substrate, the via 32 being used for electrical connection of the top and bottom sides; and also showing a cross-sectional view taken along a line extending along the via 32 on the right side.

[0373] iv. According to Figure 4a A bottom view of the second (intermediate) wafer substrate of the implantable device, on which electronic components 35, 31b and 31c are attached; and

[0374] v. according to Figure 4aA bottom view of the underside of the third (bottom) wafer substrate 27c of the implantable device, and a cross-sectional view taken along line BB. Wafer substrate 27c is a glass substrate with through-holes formed to cover (electronic) components attached to the bottom surface of the intermediate wafer substrate 27b. The cover plate 27c is welded to the bottom surface of the intermediate wafer substrate 27b at welding points 28 using a femtosecond laser welding method. This welding creates an hermetically sealed chamber between the cover plate 27c and the bottom surface of the intermediate wafer substrate 27b. Due to the hermetically sealed design, no bodily fluids can enter the chamber, and the electronic components are reliably protected from corrosion. Furthermore, the patient's body is reliably protected from any non-biocompatible materials of the biosensor throughout the lifespan of the implantable device, which is reliably encapsulated by an encapsulation device formed by three wafer substrates 27a, 27b, and 27c. In this embodiment, no part of the patient's body comes into direct contact with another material other than the biocompatible glass forming the encapsulation device.

[0375] Description of a preferred embodiment of a method for manufacturing an implantable device according to the present invention:

[0376] Each biocompatible glass layer was prepared and cleaned under cleanroom conditions.

[0377] Preferably, the packaging device uses one or more, preferably two or more, especially three or four or up to four wafer substrates to stack electrodes and biosensor films, especially electronic control devices, and, preferably, NFC antennas, and each wafer is aligned with a reference point and calibrated to the aligned stack.

[0378] Femtosecond lasers are used to pass through a controlled phase plane to enable internal splicing and welding of glass layers, thereby creating at least one hermetically sealed chamber.

[0379] The sealed chamber is preferably able to withstand 5 atmospheres (about 550 kPa) for 4 hours, with a residence time of 2 hours, and preferably, the leakage rate measured by the mass spectrometer is no more than 5 × 10⁻⁸ atm·cm³ / s.

[0380] Femtosecond lasers create micro-regions of stress, requiring unique boundary layers for each wafer design and chip geometry to allow for the sealing of multiple chambers on a single chip. This improves the overall yield of biocompatible glass wafers and enhances manufacturing efficiency. The method and design guidelines are applicable to glass wafer fabrication processes to ensure that uniform chips with sealed chambers, cut from the wafer, are protected from stress caused by the working heat-affected zone within the splicing area.

[0381] Preferably, multiple (two or more) wafers containing multiple wafer substrates for parallel fabrication of multiple implantable devices are arranged in a stacked manner, then welded at splice points or areas, and then the resulting stack of welded glass wafer substrates is cut from the stacked and welded wafers, including at least one component (electrode, electronic control device (especially microcontroller), one or more amplifiers, RFID (preferably NFC), chipset; antenna) enclosed in one or more chambers.

[0382] After these layers are soldered, a quality assessment is performed on each chip to ensure that the hermeticity and arrangement of the internal working components of the device are within performance requirements.

[0383] This laser seal allows for the creation of biocompatible sealed chambers that can be implanted for fully internal operation of electronic devices to amplify signals generated by biosensors.

[0384] In a preferred embodiment of the method for preparing a glass encapsulation device, the method includes one or more of the following steps:

[0385] - Each biocompatible glass layer is prepared and cleaned under cleanroom conditions.

[0386] The device uses up to four wafers to stack electronic devices, biosensors, NFC antennas, and biofilms, with each wafer aligned to a reference point and calibrated for an aligned stack.

[0387] - Femtosecond lasers are used to pass through a controlled phase plane to enable the internal splicing and welding of glass layers, thereby creating a sealed to hermetic chamber.

[0388] - The sealed chamber must withstand 5 atmospheres (approximately 550 kPa) for 4 hours, with a residence time of 2 hours, and the leakage rate measured by the mass spectrometer must not exceed 5 × 10⁻⁸ atm·cm³ / s.

[0389] - Femtosecond lasers create micro-regions of stress, and each wafer design and chip geometry requires a unique boundary layer to allow multiple chambers to be sealed on a single chip. This improves the overall yield of biocompatible glass wafers and enhances manufacturing efficiency. This glass wafer manufacturing process allows for the removal of stress from the single, sealed chip (a stacked wafer substrate) cut from the wafer, caused by the working heat-affected zone within the splicing area.

[0390] - After laser welding of each chamber, a chip unification process is performed, which is completed by a standard glass cutting operation, in which the glass is cut along scribing lines and the heat-affected zone.

[0391] - After these layers are soldered, a quality assessment is performed on each chip to ensure that the hermeticity and arrangement of the internal working components of the device are within performance requirements.

[0392] This laser seal allows for the inclusion of a biocompatible sealed chamber within an electronic device, which can be implanted for fully internal operation of the device to amplify signals generated by biosensors.

[0393] - The laser welding method preferably completely seals the electronic device within a glass chamber. An exemplary dimension of this chamber is 16 × 1.65 × 0.55 mm. The electronic device is connected to the aforementioned biosensor via a through-hole (also referred to as a TGV) with traced wiring on the glass surface. The TGV is hermetically sealed and allows electrical contact with the biosensor electrodes on the external surface.

[0394] Figure 6 A system 1000 for sensing at least one hormone in bodily fluids within a patient's body is shown in detail, comprising:

[0395] - A subcutaneously implantable device 1 or 21, which additionally includes a communication device 11c or 31c for wireless data communication with an external computer device 500, particularly an NFC (Near Field Communication) device, which is separate from the implantable device, and

[0396] - An external computer device 500, particularly a portable computer device, preferably a smartphone or tablet PC, includes a CPU 501 for executing computer program code, particularly the computer program code according to the invention, and a communication device 511 for wireless data communication with an implantable device, particularly an NFC device.

[0397] Preferably, when the program code (e.g., an application program) executed by the CPU on the computer device receives user input via the touch screen (not shown) of the computer device 500, data exchange is initiated, and the computer device 500 sends a request to the NFC devices 11c and 31c to transmit measurement data (preferably in encrypted form) to the computer device 500.

[0398] Preferably, measurements performed by the biosensor device 2 of the implantable device 1 are initiated when the computer device 500 sends a request to the NFC devices 11c and 31c to initiate the measurement, provided that program code (e.g., an application) executed by the CPU on the computer device receives user input via the touchscreen (not shown) of the computer device 500. Since the implantable device is preferably a battery-free device, the radio waves of the communication device 501 are preferably used to provide the energy required for measuring and / or encrypting measurement data and / or transmitting measurement data to the computer device via the wireless connection 800.

[0399] Figure 7 A schematic diagram of an implementation scheme for an electronic circuit is shown. (For example...) Figure 7 As shown, the electronic circuitry embedded within the implantable device consists of 4i commercial off-the-shelf components. These four components include a central microcontroller 5i (MCU), an ultra-low-power microcontroller with an 8-bit architecture, such as the Microchip ATTiny20 and PIC12LF device series from AZ Chandler, USA; an NFC chip tag 6i, such as the NTAG I2Cplus series from NXP Eindhoven, Netherlands; and finally, two operational amplifiers 3 with dual amplification channels, exhibiting very low input bias current (e.g., 1 pA) and ultra-low current consumption (e.g., 20 μA), such as the AD8506 series from MA Wilmington Analog Devices, USA. The maximum current consumption of the electronic circuitry 2i is set at 1 mA, with a minimum voltage level of 1.8 V, resulting in a power consumption of 1.8 mW. Three electrochemical analytes or substances can be simultaneously monitored by an available working electrode pad 9i, which is connected to three independent amplifiers 3i mounted in a transimpedance circuit topology 29i, to be monitored via, as... Figure 7 Resistor 15i in the schematic diagram converts and amplifies the ion current to a voltage equivalent level relative to the reference voltage level applied by reference electrode 28i. The output voltage signal of each amplifier 3i is then digitized at a rate of 24 samples per second and a resolution of 10 bits in three independent analog-to-digital converters (ADCs) within microcontroller 5i. An additional amplifier 4i is used during amperometric measurements to generate the aforementioned voltage reference level via a voltage divider circuit topology centered on resistors 16i and 17i, followed by signal buffering by amplifier 4i. A similar voltage divider topology is employed at the positive terminal of each transimpedance amplifier 29i to generate an internal bias voltage level that shifts the amplification baseline above the ground signal; this is achieved via resistors 18i and 19i.

[0400] Data samples generated during the digitization phase are temporarily stored in the RAM memory of the MCU 5i, digitally processed and encrypted using a lightweight security algorithm, and then digitally transmitted to the NFC chip tag 6i at a speed of 9600 bps (baud rate) using the standard I2C interface 20i protocol. In the NFC chip tag 6i, the data samples are internally stored in non-volatile memory.

[0401] Although the I2C communication protocol was originally developed by Philips Semiconductors, which covers the definition of communication signals (SDA and SCL) and specifications for transmission baud rate and byte format, the data packets (byte streams) exchanged between the MCU and the NFC chip tag are entirely custom-made here. Some of these transmitted bytes encode the sequence of actions that the MCU 5i needs to perform, such as acquiring biosignals (i.e., current), and data transmission or control parameters for the operation of the implant 1i itself. While the acquired voltage level 13i can be generated by any NFC-enabled device 8i or any RF source of the same frequency, the MCU 5i will not begin any operation until a valid command sequence is received from an authorized mobile phone running the corresponding application.

[0402] Figure 2 illustrates the operation of the biosensor device 1i. The biosensor device 1i is an implantable device and includes an encapsulation device according to the invention, made primarily of glass. For scale, the biosensor device 1i is shown positioned on top of the coin 21i. The NFC antenna 7i is shown wrapped around the perimeter of the PCB board. When the NFC interface of the mobile phone 8i is successfully powered, a small embedded light-emitting diode (LED) 11i provides visual feedback on the operation performed on the implantable side. In the presence of an external NFC field 22, internally stored and encrypted data samples are wirelessly transmitted to the external mobile phone 8i (uplink) and visualized on dedicated application software 31i installed on the phone 8i. Information regarding temperature levels is also provided via the uplink communication channel, as the microcontroller 5i also has an internal temperature indicator built into the die. Through the same process, AC radio frequency waves 22i generated by the mobile phone 8i are continuously captured by the internal NFC antenna 7i. Antenna 7i is made of 20 turns of 0.15 mm thick enameled copper wire and is wound around the outer boundary of circuit board 12i, having dimensions of, for example, 6 mm × 12 mm, and is converted to an equivalent DC level 13 (V) by the NFC chip tag 6i itself before being made usable in the rest of the embedded electronic device. 采集 In the opposite direction of communication (or downlink), specific data 10i (e.g., user parameters) can be sent from the same mobile phone application software 31ii to set the ID number to the implantable biosensor device 1i, such as the device operation and control of the amperometric measurement method.

[0403] exist Figure 9 The diagram schematically illustrates the layout of the rigid PCB back side of the biosensor device, and... Figure 10The image shows a 3D schematic layout of the front side of a rigid PCB for a biosensor device. Physically, the described electronic schematic of the implantable device is translated into printed circuit board (PCB) technology, where protected trace conductors 24i and through-layer vias 25i are vertically interconnected pathways, i.e., conductive paths or holes connecting different layers of, for example, a PCB or IC, thereby providing current flow through the board layers to transmit electrical signals between different chipsets, resistors 15i-19i, 30i, capacitor 14i, and NFC antenna 7i. Exposed electronic pads 23i, made by a Ni / Au surface finishing process, are used to solder components to the PCB 12i substrate, or uncovered electronic pads 27i are left to allow connection to programming pins of the MCU 5i or to allow electrochemical functions of the electrochemical sensor, such as… Figure 9 and 10 As shown. Copper traces 24i are placed on the top and bottom layers of PCB 12i, while component assembly is performed only on the top layer. Rigid 12i (e.g., laminated copper plus epoxy in an FR-4 circuit board) or flexible 12i (e.g., polyimide film base) can be used to manufacture the PCB 8i.

[0404] Any method disclosed herein includes one or more steps or actions for performing the method. These method steps and / or actions are interchangeable. In other words, the order and / or use of a particular step and / or action may be modified unless the correct operation of the implementation requires a specific order of steps or actions. Furthermore, within the scope of this disclosure, a subroutine of the method described herein, or only a portion thereof, may be a separate method. In other words, some methods may include only a subset of steps described in a more detailed method.

[0405] Throughout this specification, references to "implementation," "this implementation," or "preferred configuration" mean that a particular feature, structure, or characteristic described in connection with that implementation is included in at least one implementation. Therefore, phrases or variations thereof referenced throughout this specification do not necessarily refer to the same implementation.

[0406] The use of the term "first" in the claims regarding a feature or element does not necessarily imply the existence of a second or additional such feature or element. Changes may be made to the details of the above embodiments without departing from the basic principles of this disclosure.

[0407] Furthermore, those skilled in the art who benefit from this disclosure will understand that, for the purpose of simplification, multiple features are sometimes combined in a single embodiment, drawing, or description thereof in the above description of the embodiments. However, this approach to disclosure should not be construed as reflecting an intention in any claim to require more features than those expressly stated in that claim. Rather, as reflected in the following claims, the inventive aspect lies in a combination of fewer features than all features of any single foregoing disclosed embodiment. Therefore, the claims are thus expressly incorporated into this specification, each claim being a separate embodiment in itself. This disclosure includes all permutations and combinations of the independent claims and their dependent claims.

Claims

1. A biosensor device for sensing at least one hormone, comprising: (a) at least one metal electrode surface; and (b) A biosensor membrane comprising at least one aptamer attached to the surface of the metal electrode, wherein the aptamer: (i) capable of binding to at least one of the hormones; and (ii) Modified with one or more functional groups for attaching the at least one aptamer to the surface of the metal electrode.

2. The biosensor device according to claim 1, configured to sense at least one hormone in bodily fluids, particularly interstitial fluid.

3. The biosensor device according to any one of the preceding claims, wherein the at least one hormone is selected from the group consisting of estradiol, luteinizing hormone (LH), progesterone, and any combination thereof.

4. The biosensor device according to any one of the preceding claims, wherein the biosensor device further comprises a carrier, particularly a glass carrier, such as a wafer substrate, for supporting the surface of the at least one metal electrode.

5. The biosensor device according to any one of the preceding claims, wherein the aptamer is a single-stranded nucleic acid molecule, preferably a DNA or RNA molecule.

6. The biosensor device according to any one of the preceding claims, wherein the aptamer is about 25 to 70 nucleotides in length, preferably about 30 to about 65 nucleotides.

7. The biosensor device according to any one of the preceding claims, wherein the functional group for attaching the at least one aptamer to the surface of the metal electrode is a thiol group, which is preferably present at the end of the aptamer.

8. The biosensor device according to any one of the preceding claims, wherein the aptamer is a single-stranded nucleic acid molecule, preferably DNA or a DNA molecule, and the functional group for attaching the at least one aptamer to the surface of the metal electrode is present at the 3' end or the 5' end, preferably the 5' end.

9. The biosensor device according to any one of the preceding claims, wherein the aptamer is capable of binding to at least one hormone selected from the group consisting of estradiol, luteinizing hormone (LH), and progesterone.

10. The biosensor device according to any one of the preceding claims, wherein the aptamer is selected from one or more of the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO:

4.

11. The biosensor device according to any one of claims 1 to 10, for diagnostic purposes, particularly for sensing at least one hormone, and more particularly for monitoring hormones during assisted reproductive technology (ART), during menopause, in hormonal disorders such as polycystic ovary syndrome (PCOS), in endometriosis, and / or during hormone therapy.

12. Implantable devices for implantation, particularly subcutaneous implantation into animals or humans, including • The biosensor device according to any one of the preceding claims, and • An electronic control device for controlling the biosensor device, particularly for generating, collecting, and preferably encrypting measurement data sensed by the biosensor device.

13. A system for sensing at least one hormone, comprising: The implantable device of claim 12 further includes a communication means for wireless data communication with an external computer device, particularly an NFC (Near Field Communication) device, said external computer device being separate from the implantable device, and The external computer device includes a communication device, particularly an NFC device, for wireless data communication with the implantable device.

14. The biosensor device according to any one of claims 1 to 11 is used for in vitro diagnostics, particularly for sensing at least one hormone.

15. A method for operating a biosensor device according to any one of claims 1 to 11, an implantable device according to claim 12, or a system according to claim 13, comprising the following steps: (a) Contact the biosensor membrane with a body fluid containing at least one of the hormones, particularly interstitial fluid; (b) Binding the at least one hormone to the at least one aptamer of the biosensor membrane; as well as (c) Detecting the binding of the at least one hormone by electrically controlling the surface of the at least one metal electrode.

Images