1. A system for extracting protein from human nail samples, comprising: a lysis buffer chamber for dissolving nail samples, wherein the lysis buffer chamber further comprise a lysis buffer solution; a sample transfer mechanism for transferring dissolved nail samples into separate tubes; a filtration unit for separating protein solutions from undissolved sample components through centrifugation; a pulverization apparatus for pulverizing undissolved sample components using a magnetic stirrer; a temperature control unit maintaining samples at specified temperatures ranging from about 40 degree Celcius to about 90 degree Celcius, during processing steps; a supernatant separation mechanism for separating supernatants from undissolved samples by centrifugation; an acetone precipitation unit for precipitating sample fractions; and a sample resuspension chamber for resuspending precipitated sample fractions in a suitable sample buffer.

2. The system of claim 1, wherein the lysis buffer solution further comprises: from about 0.1M to about 0.2M NaOH; Sodium Dodecyl Sulphate (SDS) at 1% by weight; ß-mercaptoethanol (ß ME) at 2% by weight; and 0.01 M Ethylenediaminetetraacetic Acid (EDTA).

3. The system of claim 1, further comprising a data acquisition and control module for monitoring and controlling the sample preparation process, including temperature regulation and timing of sample processing steps.

4. The system of claim 1, further comprising a user interface for inputting sample-specific parameters and controlling the sample preparation process.

5. A method for extracting protein from human nail samples, comprising: dissolving nail samples in a lysis buffer solution; transferring dissolved nail samples into separate tubes by using a sample transfer mechanism; separating protein solutions from undissolved sample components through centrifugation using a filtration unit; pulverizing undissolved sample components using a magnetic stirrer; separating supernatants from undissolved samples using a supernatant separation mechanism; precipitating sample fractions using acetone precipitation; and resuspending precipitated sample fractions in a suitable sample buffer.

6. The method of claim 5, further comprising the step of maintaining samples at specified temperatures ranging from about 40-degree Celsius to about 90-degree Celsius, during processing steps using a temperature control unit.

7. The method of claim 5, wherein sample-specific parameters are inputted and the sample preparation process is controlled using a user interface.

8. The method of claim 5, further comprising the step of resuspending the pelletized proteins in a suitable sample buffer after the acetone precipitation.

9. The method of claim 5, further comprising the step of repeating the pulverization, centrifugation, and supernatant separation steps to maximize the yield of extracted proteins.

10. The method of claim 5, further comprising step of characterizing extracted protein from human nail samples by using characterization techniques selected from Fourier-Transform Infrared (FTIR) spectroscopy, X-ray Diffraction (XRD), Scanning Electron Microscopy - Energy Dispersive X-ray Spectroscopy (SEM-EDX), Inductively Coupled Plasma (ICP) analysis and Thermogravimetric (TG) analysis. PURIFICATION METHOD FOR KERATIN EXTRACTION FROM HUMAN NAIL WASTES AND DETECTION OF HEAVY METALS Abstract The present disclosure presents a system designed for the efficient extraction of proteins from nail samples. The system comprises a lysis buffer chamber housing a specialized lysis buffer solution engineered for dissolving nail samples effectively. Following dissolution, a sample transfer mechanism is employed to transfer the dissolved samples into separate tubes. The system further incorporates a filtration unit, utilizing centrifugation to segregate protein solutions from the undissolved sample components. Any undissolved sample components are then subjected to a pulverization apparatus employing a magnetic stirrer to ensure thorough pulverization. A temperature control unit within the system maintains the samples at specified temperatures, ranging from about 40 degrees Celsius to about 90 degrees Celsius, throughout various processing steps to ensure optimal conditions for protein extraction. The system also encompasses a supernatant separation mechanism, which employs centrifugation to segregate supernatants from undissolved samples. An acetone precipitation unit is integrated for precipitating sample fractions, which are subsequently transferred to a sample resuspension chamber where the precipitated sample fractions are resuspended in a suitable sample buffer for further analysis or utilization. The disclosure provides a comprehensive, streamlined, and effective approach for protein extraction from nail samples, thus paving the way for enhanced utilization of such biological materials in a variety of scientific and industrial applications. , Claims:Claims :

1. A system for extracting protein from human nail samples, comprising: a lysis buffer chamber for dissolving nail samples, wherein the lysis buffer chamber further comprise a lysis buffer solution; a sample transfer mechanism for transferring dissolved nail samples into separate tubes; a filtration unit for separating protein solutions from undissolved sample components through centrifugation; a pulverization apparatus for pulverizing undissolved sample components using a magnetic stirrer; a temperature control unit maintaining samples at specified temperatures ranging from about 40 degree Celcius to about 90 degree Celcius, during processing steps; a supernatant separation mechanism for separating supernatants from undissolved samples by centrifugation; an acetone precipitation unit for precipitating sample fractions; and a sample resuspension chamber for resuspending precipitated sample fractions in a suitable sample buffer.

2. The system of claim 1, wherein the lysis buffer solution further comprises: from about 0.1M to about 0.2M NaOH; Sodium Dodecyl Sulphate (SDS) at 1% by weight; ß-mercaptoethanol (ß ME) at 2% by weight; and 0.01 M Ethylenediaminetetraacetic Acid (EDTA).

3. The system of claim 1, further comprising a data acquisition and control module for monitoring and controlling the sample preparation process, including temperature regulation and timing of sample processing steps.

4. The system of claim 1, further comprising a user interface for inputting sample-specific parameters and controlling the sample preparation process.

5. A method for extracting protein from human nail samples, comprising: dissolving nail samples in a lysis buffer solution; transferring dissolved nail samples into separate tubes by using a sample transfer mechanism; separating protein solutions from undissolved sample components through centrifugation using a filtration unit; pulverizing undissolved sample components using a magnetic stirrer; separating supernatants from undissolved samples using a supernatant separation mechanism; precipitating sample fractions using acetone precipitation; and resuspending precipitated sample fractions in a suitable sample buffer.

6. The method of claim 5, further comprising the step of maintaining samples at specified temperatures ranging from about 40-degree Celsius to about 90-degree Celsius, during processing steps using a temperature control unit.

7. The method of claim 5, wherein sample-specific parameters are inputted and the sample preparation process is controlled using a user interface.

8. The method of claim 5, further comprising the step of resuspending the pelletized proteins in a suitable sample buffer after the acetone precipitation.

9. The method of claim 5, further comprising the step of repeating the pulverization, centrifugation, and supernatant separation steps to maximize the yield of extracted proteins.

10. The method of claim 5, further comprising step of characterizing extracted protein from human nail samples by using characterization techniques selected from Fourier-Transform Infrared (FTIR) spectroscopy, X-ray Diffraction (XRD), Scanning Electron Microscopy - Energy Dispersive X-ray Spectroscopy (SEM-EDX), Inductively Coupled Plasma (ICP) analysis and Thermogravimetric (TG) analysis.

Description:PURIFICATION METHOD FOR KERATIN EXTRACTION FROM HUMAN NAIL WASTES AND DETECTION OF HEAVY METALS
Field of the Invention
[0001] The present disclosure relates generally to the field of biochemical processing and, more particularly, to a system and method for the extraction of keratin protein from human nail wastes and the subsequent detection of heavy metals. The disclosure is aimed at not only efficiently extracting valuable keratin proteins from human nail wastes, which are typically discarded, but also analyzing the extracted keratin for the presence of heavy metals, thus contributing to both environmental sustainability and human health monitoring.
Background
[0002] The background description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.
[0003] The development of efficient and environmentally sustainable methods for the extraction and purification of biologically derived materials has been a long-standing endeavor in the field of biotechnology. Among such materials, keratin, a fibrous protein forming the main structural constituent of hair, feathers, hoofs, claws, horns, and nails, has garnered considerable attention due to its robust physical properties and its potential utilization in various industrial applications including medical, cosmetic, and material science sectors.
[0004] Historically, the primary sources of keratin for commercial and research purposes have been animal-derived, such as from wool and feathers. However, human nail wastes present a viable and relatively unexplored source of keratin, given the ubiquitous nature of nail clippings and their general disposal as waste. Despite this potential, conventional methods of keratin extraction from human nails have been less explored and often entail harsh chemical treatments or prolonged enzymatic digestion, which could lead to the degradation of keratin or result in low yields.
[0005] Furthermore, the efficient detection of heavy metals in keratinized matrices such as nails is of paramount importance, as the presence of heavy metals could reflect environmental or occupational exposures to toxic substances. Prior art discloses various methods for heavy metal detection, often relying on sophisticated analytical techniques such as inductively coupled plasma mass spectrometry (ICP-MS) or atomic absorption spectroscopy (AAS). However, said methods require standalone processes post keratin extraction and purification, necessitating additional processing steps and potentially altering the native state of the keratin matrix, which may hinder the accuracy of heavy metal quantification.
[0006] Moreover, some prior art describes keratin extraction methods from animal sources. For instance, a prior art example discloses a method of extracting keratin from feathers using an ionic liquid solvent system. Another example, discusses a method of extracting keratin from wool through controlled oxidation followed by enzymatic hydrolysis. However, said examples do not address the unique challenges posed by human nail wastes nor do they discuss the simultaneous detection of heavy metals.
[0007] Therefore, a more streamlined and integrated method for both keratin extraction from human nail wastes and heavy metal detection within the extracted keratin is highly desirable. Such a method would ideally address the challenges posed by the resilient nature of human nail keratin, while allowing for efficient and accurate heavy metal detection, thereby facilitating broader research and commercial applications of human-derived keratin, and enabling a more comprehensive understanding and monitoring of heavy metal exposures in human populations.
[0008] The new purification method for keratin extraction from human nail wastes and detection of heavy metals as proposed in the study under consideration, seeks to address the aforementioned limitations in the prior art, by introducing an efficient, and integrated approach to not only extract keratin from human nail wastes but also to detect and quantify heavy metal residues within the extracted keratin, all within a streamlined and coherent processing framework.
[0009] All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.
Summary
[00010] Various objects, features, and advantages of the disclosed subject matter can be more fully appreciated with reference to the following detailed description of the disclosed subject matter when considered in connection with the following drawings, in which like reference numerals identify like elements.
[00011] The present disclosure relates generally to the field of biochemical processing and, more particularly, to a system and method for the extraction of keratin protein from human nail wastes and the subsequent detection of heavy metals. The disclosure is aimed at not only efficiently extracting valuable keratin proteins from human nail wastes, which are typically discarded, but also analyzing the extracted keratin for the presence of heavy metals, thus contributing to both environmental sustainability and human health monitoring.
[00012] The system designed for the extraction of proteins from human nail samples represents a sophisticated and integrated approach to address the challenges associated with the unique biological material. The comprehensive system consists of several key components, each playing a crucial role in ensuring the efficient and effective extraction of proteins while maintaining their integrity.
[00013] The system features a lysis buffer chamber, where the initial step of dissolving nail samples occurs. The chamber houses a precisely formulated lysis buffer solution, carefully crafted to create an environment conducive to the dissolution of nail samples. The lysis buffer solution includes essential components such as sodium hydroxide (NaOH), sodium dodecyl sulfate (SDS), ß-mercaptoethanol (ß ME), and ethylenediaminetetraacetic acid (EDTA). Said components work in concert to break down cell structures, solubilize proteins, and prevent interference from metal ions.
[00014] Following the dissolution of nail samples, a sample transfer mechanism facilitates the transfer of the resulting solution into separate tubes. The step ensures the isolation of individual samples, preventing cross-contamination and ensuring the integrity of the extracted proteins.
[00015] The subsequent step involves the use of a filtration unit for the separation of protein solutions from undissolved sample components. Centrifugation is employed to achieve the separation, with denser components settling at the bottom while protein-rich supernatants remain above.
[00016] To address challenging nail samples with resistant undissolved components, the system incorporates a pulverization apparatus equipped with a magnetic stirrer. Mechanical agitation is applied to further break down said components, enhancing protein release.
[00017] Precise temperature control is a critical aspect of the system, with a temperature control unit ensuring that samples are maintained within specified temperature ranges, typically ranging from 40 to 90 degrees Celsius. Temperature regulation is pivotal, influences various biochemical reactions during the extraction process.
[00018] A supernatant separation mechanism is employed to further refine the extracted protein fractions. The mechanism involves subjecting the supernatants to additional centrifugation, ensuring the removal of any remaining impurities or particles.
[00019] The system's capabilities extend to protein fraction precipitation through an acetone precipitation unit. Acetone is added to the protein solution, causing proteins to precipitate out of the solution. The step concentrates and purifies the protein fractions, removing unwanted contaminants.
[00020] Lastly, a sample resuspension chamber allows for the gentle and controlled resuspension of the precipitated protein fractions in a suitable sample buffer. The step is vital to maintain the solubility and stability of the proteins, making them ready for downstream applications.
[00021] Complementing said core components, the system includes a data acquisition and control module for precise monitoring and control of the sample preparation process. The system oversees temperature regulation, timing of processing steps, and error detection, ensuring reproducibility and reliability.
[00022] Furthermore, a user interface simplifies the system's operation by allowing researchers to input sample-specific parameters and control the entire sample preparation process. The user-friendly interface enhances accessibility and adaptability, accommodating both novice and experienced users.
[00023] Hence, the system represents a holistic solution for the extraction of proteins from human nail samples. The integration of lysis buffer chemistry, mechanical agitation, temperature control, and precise separation techniques ensures the generation of high-quality protein extracts. Researchers can rely on the system to efficiently process nail samples, paving the way for a multitude of applications in the fields of proteomics, biomarker discovery, and disease research.
[00024] The method for extracting proteins from human nail samples described herein offers a comprehensive and meticulously orchestrated process to obtain high-quality protein fractions from the challenging biological material.
[00025] The method begins with the dissolution of nail samples in a precisely formulated lysis buffer solution. The solution is designed to create an environment conducive to the release of proteins from the nail samples. The carefully balanced composition includes key components such as sodium hydroxide (NaOH), sodium dodecyl sulfate (SDS), ß-mercaptoethanol (ß ME), and ethylenediaminetetraacetic acid (EDTA). Said components work synergistically to break down cellular structures and solubilize proteins.
[00026] Following dissolution, the method employs a sample transfer mechanism to transfer the dissolved nail samples into separate tubes. The step ensures the isolation of individual samples and prevents cross-contamination, preserving the integrity of the extracted proteins.
[00027] The subsequent centrifugation step, executed through a filtration unit, separates protein solutions from undissolved sample components. Centrifugal force drives denser components to the tube's bottom while retaining protein-rich supernatants, a crucial step in obtaining purified protein fractions.
[00028] To address stubborn undissolved components, the method incorporates a pulverization apparatus equipped with a magnetic stirrer. Mechanical agitation complements chemical dissolution, enhancing the release of proteins from the nail samples.
[00029] Precise temperature control is central to the method, maintained by a temperature control unit. The unit ensures that samples remain within specified temperature ranges, typically from 40 to 90 degrees Celsius, influencing critical biochemical reactions during the extraction process. The method's refinement continues with a supernatant separation mechanism. Additional centrifugation removes any remaining impurities or particles from the supernatants, further enhancing protein purity.
[00030] Protein fraction precipitation using acetone follows, concentrating and purifying the protein fractions by causing them to precipitate from the solution. The step effectively removes salts and contaminants. Finally, the method concludes with the resuspension of the pelletized proteins in a suitable sample buffer. Said resuspension ensures the proteins' solubility and stability, rendering them ready for downstream applications.
[00031] The method's versatility is underscored by optional steps that allow for maximum protein yield. Researchers can repeat the pulverization, centrifugation, and supernatant separation steps when dealing with particularly challenging nail samples.
[00032] Additionally, the method opens doors for thorough protein characterization, using techniques such as Fourier-Transform Infrared (FTIR) spectroscopy, X-ray Diffraction (XRD), Scanning Electron Microscopy - Energy Dispersive X-ray Spectroscopy (SEM-EDX), Inductively Coupled Plasma (ICP) analysis, and Thermogravimetric (TG) analysis. Said techniques provide valuable insights into the structural and chemical properties of the extracted proteins, expanding the range of potential applications in research and diagnostics.
[00033] In essence, the method represents a systematic and meticulous approach to protein extraction from human nail samples. The incorporation of lysis buffer chemistry, mechanical agitation, temperature control, and purification techniques ensures the generation of high-quality protein extracts. Researchers can rely on the method to efficiently process nail samples, setting the stage for advancements in proteomics, biomarker discovery, and disease research.
Brief Description of the Drawings
[00034] The features and advantages of the present disclosure would be more clearly understood from the following description taken in conjunction with the accompanying drawings in which:
[00035] FIG. 1 represents an architectural outline of a system for extracting protein from human nail samples, according to some embodiments of the present disclosure.
[00036] FIG. 2 depicts an exemplary detailed schematic flow diagram of a method for extracting protein from human nail samples, according to some embodiments of the present disclosure.
Detailed Description
[00037] The following is a detailed description of exemplary embodiments to illustrate the principles of the invention. The embodiments are provided to illustrate aspects of the invention, but the invention is not limited to any embodiment. The scope of the invention encompasses numerous alternatives, modifications and equivalent; it is limited only by the claims.
[00038] In view of the many possible embodiments to which the principles of the present discussion may be applied, it should be recognized that the embodiments described herein with respect to the drawing figures are meant to be illustrative only and should not be taken as limiting the scope of the claims. Therefore, the techniques as described herein contemplate all such embodiments as may come within the scope of the following claims and equivalents thereof.
[00039] The detailed description is described with reference to the accompanying figures. In the figures, the left-most digit(s) of a reference number identifies the figure in which the reference number first appears. The use of the same reference numbers in different instances in the description and the figures may indicate similar or identical items.
[00040] Pursuant to the "Detailed Description" section herein, whenever an element is explicitly associated with a specific numeral for the first time, such association shall be deemed consistent and applicable throughout the entirety of the "Detailed Description" section, unless otherwise expressly stated or contradicted by the context.
[00041] The present disclosure relates generally to the field of biochemical processing and, more particularly, to a system and method for the extraction of keratin protein from human nail wastes and the subsequent detection of heavy metals. The disclosure is aimed at not only efficiently extracting valuable keratin proteins from human nail wastes, which are typically discarded, but also analyzing the extracted keratin for the presence of heavy metals, thus contributing to both environmental sustainability and human health monitoring.
[00042] Pursuant to the "Detailed Description" section herein, whenever an element is explicitly associated with a specific numeral for the first time, such association shall be deemed consistent and applicable throughout the entirety of the "Detailed Description" section, unless otherwise expressly stated or contradicted by the context.
[00043] Presented herein a system 100 for extracting protein from human nail samples offers a comprehensive solution for isolating proteins from said challenging biological matrices. The system 100 integrates various components and techniques to efficiently process nail samples and obtain protein fractions suitable for further analysis. The comprehensive discussion delves into the system's key components, their functions, and their roles in the protein extraction process. Furthermore, explored about the lysis buffer solution, the composition, and significance. Additionally, disclosure touches upon the system's data acquisition and control module and user interface, highlighting their importance in ensuring precision and ease of operation.
[00044] Pictorial elucidation of FIG. 1, illustrates an architectural setup of the system 100 that can comprise a lysis buffer chamber 102, a sample transfer mechanism 104, a filtration unit 106, a pulverization apparatus 108, a temperature control unit 110, a supernatant separation mechanism 112, an acetone precipitation unit 114, a sample resuspension chamber 116 and other known elements thereof. A person ordinarily skilled in art would prefer those elements or components of the system 100, to be functionally or operationally coupled with each other, in accordance with the embodiments of present disclosure.
[00045] In an embodiment, the lysis buffer chamber serves as the initial stage in the protein extraction process. Said chamber’s primary purpose is to dissolve nail samples, thereby breaking down the cellular structures and releasing proteins into solution. The efficiency of the step is crucial for obtaining high-quality protein extracts. The lysis buffer solution employed in the system is a critical component. The lysis buffer solution can comprise several key reagents that facilitate nail sample dissolution and protein solubilization.
[00046] Referring to the preceding embodiment, said key reagents include sodium Hydroxide (NaOH), present in concentrations ranging from about 0.1M to 0.2M. NaOH serves as an alkaline agent, helping to disrupt cell membranes and release proteins from the nail samples. Sodium Dodecyl Sulphate (SDS) can be added at a concentration of 1% by weight, SDS is a powerful detergent that aids in the solubilization of membrane proteins by disrupting hydrophobic interactions.
[00047] Referring to the preceding embodiment, further, ß-mercaptoethanol (ß ME) included at 2% by weight, ß ME is a reducing agent that helps break disulfide bonds, further assisting in protein solubilization. Ethylenediaminetetraacetic Acid (EDTA) added at a concentration of 0.01M, EDTA is a chelating agent that sequesters divalent metal ions, which can interfere with various enzymatic reactions during protein extraction. Together, said components create an environment within the lysis buffer chamber that is conducive to the dissolution of nail samples and the release of proteins into solution. The careful formulation of the lysis buffer solution is essential for efficient protein extraction, ensuring the preservation of protein structure and function during the extraction process.
[00048] In an exemplary embodiment, post the nail samples have been dissolved in the lysis buffer, the next step involves transferring the resulting solution into separate tubes. Said step is critical in the process as the step allows for the subsequent separation and processing of the sample components. The sample transfer mechanism facilitates the precise and efficient transfer of the dissolved nail samples into designated tubes. Proper handling at the stage ensures that all subsequent steps, including centrifugation and supernatant separation, can be carried out effectively.
[00049] Once the dissolved nail samples are in separate tubes, the filtration unit comes into play. The filtration unit can be configured to separate the protein solutions from undissolved sample components. The separation is achieved through centrifugation, a process that relies on the differential sedimentation rates of the various components in the sample. During centrifugation, the tubes containing the dissolved nail samples are subjected to high-speed rotation. The centrifugal force causes the denser components, including undissolved material and cell debris, to sediment at the bottom of the tubes, while the protein solutions remain in the supernatant. The step is pivotal in isolating the proteins of interest from unwanted impurities and cellular debris. The separated supernatants are rich in protein content and can be further processed to obtain purified protein fractions.
[00050] In some cases, despite the use of a well-formulated lysis buffer, certain nail samples may contain stubborn undissolved components. To address the issue, the system incorporates a pulverization apparatus equipped with a magnetic stirrer. The magnetic stirrer generates mechanical agitation within the tubes, assisting in the breakdown of undissolved material and enhancing the release of proteins into solution. The step is especially valuable when dealing with challenging nail samples that may have resistant tissue structures. The combination of chemical lysis and mechanical agitation

provided by the pulverization apparatus ensures comprehensive protein extraction from the nail samples, leaving no valuable protein behind.
[00051] In an exemplary embodiment, the temperature control plays a pivotal role in maintaining the integrity of the protein samples throughout the extraction process. The system features a temperature control unit that ensures the nail samples are maintained at specified temperatures ranging from about 40 degrees Celsius to about 90 degrees Celsius during processing steps. Maintaining precise temperatures is crucial because temperature can influence various biochemical reactions, including protein solubility and enzyme activity. For instance, elevated temperatures can enhance the effectiveness of enzymatic reactions involved in protein solubilization, while lower temperatures may be necessary for specific downstream applications. The temperature control unit enables researchers to tailor the protein extraction process to the unique requirements of their experiments, ensuring the preservation of protein quality and functionality.
[00052] In an exemplary embodiment, following the initial centrifugation step, which separates the protein solutions from undissolved sample components, the system employs a supernatant separation mechanism for further refinement. The mechanism involves subjecting the tubes containing the separated supernatants to additional centrifugation. The purpose of the second centrifugation step is to ensure complete separation of any residual particles or impurities that may still be present in the supernatant. By carefully controlling the centrifugation parameters, including speed and duration, the system ensures that the supernatants are free from any remaining debris, resulting in protein solutions of high purity.
[00053] To concentrate and purify the protein fractions obtained from the supernatants, the system incorporates an acetone precipitation unit. The step involves adding acetone to the protein solution, which causes the proteins to precipitate out of solution. Acetone precipitation is a widely used method for concentrating proteins and removing salts and other contaminants. Acetone precipitation relies on the principle that proteins become less soluble in a high-concentration organic solvent like acetone. As a result, the proteins aggregate and can be easily recovered through centrifugation. The acetone precipitation unit ensures that the protein fractions are concentrated and purified, making them suitable for a variety of downstream applications such as gel electrophoresis, mass spectrometry, or protein quantification.
[00054] In an exemplary embodiment, once the protein fractions have been precipitated and collected, they need to be resuspended in a suitable sample buffer for storage or further analysis. The sample resuspension chamber is designed for the purpose. The chamber allows for the gentle and controlled resuspension of the precipitated protein fractions. Careful handling at the stage is crucial to prevent denaturation or aggregation of the proteins. The choice of sample buffer used for resuspension can also influence the stability and solubility of the proteins, making an important consideration. The system ensures that the resuspension process is carried out efficiently, resulting in protein samples ready for analysis or storage, while maintaining their structural and functional integrity.
[00055] In an exemplary embodiment, to ensure precision and reproducibility in the protein extraction process, the system is equipped with a data acquisition and control module. The module serves as the brain of the system, overseeing and regulating various aspects of the sample preparation process. Key functions of the data acquisition and control module can include monitoring and controlling temperature. As mentioned earlier, temperature control is crucial for the success of protein extraction. The module continuously monitors and adjusts the temperature within the system, ensuring that the specified temperature ranges are maintained throughout the process.
[00056] Referring to the preceding embodiment, said module controls the timing of critical steps in the protein extraction process, such as centrifugation durations and the operation of the pulverization apparatus. Precise timing is essential for achieving optimal results. Similarly, the module has built-in error detection capabilities. If any issues or anomalies are detected during the sample preparation process, can generate alerts or notifications to ensure that corrective actions are taken promptly.
[00057] Referring to the preceding embodiment, the module records data related to the sample preparation process, including temperature profiles, processing times, and any user-defined parameters. The data can be invaluable for quality control, troubleshooting, and process optimization. Hence, the data acquisition and control module enhance the reliability and repeatability of protein extraction, making a valuable component of the system for researchers seeking consistent and high-quality results.
[00058] In an exemplary embodiment, to make the system user-friendly and accessible to researchers, the system features a user interface that allows for the input of sample-specific parameters and control over the sample preparation process. The user interface serves as the means through which researchers interact with and operate the system. For instance, researchers can input sample-specific parameters, such as the type of nail sample, desired extraction temperature, and processing times. The customization ensures that the system can be tailored to the unique characteristics of each sample. The user interface enables researchers to initiate and halt the sample preparation process as needed. The control is essential for flexibility and adaptability during experimentation.
[00059] Referring to the preceding embodiment, the researchers can monitor the progress of the sample preparation process in real time through the user interface. The feature provides visibility into the status of the system and allows for immediate intervention if required. Further, data generated during the sample preparation process, such as temperature profiles and processing times, can be retrieved through the user interface for analysis and documentation. By providing researchers with a user-friendly interface, the system ensures that both novice and experienced users can effectively operate the equipment and conduct protein extraction experiments with ease and precision.
[00060] Referring to one or more preceding embodiments, the system 100 for extracting protein from human nail samples is a sophisticated and versatile platform designed to address the unique challenges posed by said biological matrices. The key components, including the lysis buffer chamber, filtration unit, pulverization apparatus, temperature control unit, supernatant separation mechanism, acetone precipitation unit, and sample resuspension chamber, work in tandem to achieve efficient protein extraction while preserving the integrity of the protein samples.
[00061] Referring to one or more preceding embodiments, the carefully formulated lysis buffer solution, containing NaOH, SDS, ß ME, and EDTA, plays a pivotal role in nail sample dissolution and protein solubilization. Meanwhile, the data acquisition and control module, coupled with the user interface, ensure precise control over the sample preparation process, monitoring of critical parameters, and ease of operation. The comprehensive system empowers researchers to extract high-quality protein fractions from human nail samples, opening up possibilities for a wide range of downstream applications, including proteomics, biomarker discovery, and disease research.
[00062] Elucidated herein the present disclosure that relates to a method 200 for extracting proteins from human nail samples. The method 200 involves a series of steps that ensure efficient extraction and preservation of the extracted proteins. Diagrammatic depiction of FIG. 2, represents a flow diagram of the method 200 that comprise steps of (at step 202) dissolving nail samples in a lysis buffer solution, (at step 204) transferring dissolved nail samples into separate tubes by using a sample transfer mechanism, (at step 206) separating protein solutions from undissolved sample components through centrifugation using a filtration unit, (at step 208) pulverizing undissolved sample components using a magnetic stirrer, (at step 210) separating supernatants from undissolved samples using a supernatant separation mechanism, (at step 212) precipitating sample fractions using acetone precipitation, and (at step 214) resuspending precipitated sample fractions in a suitable sample buffer.
[00063] Referring to the preceding embodiment, the method 200 begins with the dissolution of nail samples in a lysis buffer solution. The lysis buffer solution is carefully formulated to create an environment conducive to the dissolution of nail samples and the release of proteins into solution. Following the dissolution step, the method involves transferring the dissolved nail samples into separate tubes using a sample transfer mechanism. The step ensures that the subsequent processing steps can be carried out efficiently without cross-contamination.
[00064] In yet another embodiment, the next step in the method 200 is the separation of protein solutions from undissolved sample components. Said separation is achieved through centrifugation using a filtration unit. During centrifugation, the tubes containing the dissolved nail samples are subjected to high-speed rotation, leading to the sedimentation of denser components (undissolved material and cell debris) at the bottom of the tubes, while the protein solutions remain in the supernatant.
[00065] In yet another embodiment, in cases where certain nail samples contain stubborn undissolved components, a pulverization apparatus equipped with a magnetic stirrer is employed. The magnetic stirrer generates mechanical agitation within the tubes, aiding in the breakdown of undissolved material and enhancing the release of proteins into solution. To further refine the protein extraction process, the method includes a supernatant separation mechanism. The mechanism involves subjecting the tubes containing the separated supernatants to additional centrifugation. The purpose of the second centrifugation step is to ensure complete separation of any residual particles or impurities that may still be present in the supernatant.
[00066] In yet another embodiment, the method also incorporates acetone precipitation to concentrate and purify the protein fractions obtained from the supernatants. Acetone is added to the protein solution, causing the proteins to precipitate out of solution. The step helps remove salts and other contaminants. The precipitated proteins can be easily collected through centrifugation. Following acetone precipitation, the pelletized proteins are resuspended in a suitable sample buffer. The step ensures that the proteins are properly suspended and can be readily used for further analysis or storage. The choice of sample buffer used for resuspension can influence the stability and solubility of the proteins.
[00067] In yet another embodiment, in situations where maximum protein yield is desired, the method allows for the repetition of the pulverization, centrifugation, and supernatant separation steps. The iterative approach can help extract additional proteins from nail samples, particularly those with challenging tissue structures. The extracted proteins from human nail samples can be characterized using various techniques to assess their quality and composition.
[00068] Said characterization techniques may include Fourier-Transform Infrared (FTIR) spectroscopy, X-ray Diffraction (XRD), Scanning Electron Microscopy - Energy Dispersive X-ray Spectroscopy (SEM-EDX), Inductively Coupled Plasma (ICP) analysis, and Thermogravimetric (TG) analysis. Said techniques provide valuable insights into the structural and chemical properties of the extracted proteins, making them suitable for a wide range of downstream applications in research and diagnostics.
[00069] Referring to one or more preceding embodiments, the disclosed method 200 provides a comprehensive approach to protein extraction from human nail samples. The method 200 ensures efficient dissolution of nail samples, separation of protein solutions from undissolved components, and purification of protein fractions. The use of a well-formulated lysis buffer solution, sample transfer mechanisms, centrifugation, mechanical agitation, supernatant separation, acetone precipitation, and resuspension in a sample buffer collectively enable researchers to obtain high-quality protein extracts for further analysis and characterization.
[00070] Further, while operations are depicted in a particular order, this should not be understood as requiring that such operations be performed in the particular order shown or in sequential order, or that all illustrated operations be performed, to achieve desirable results. In certain circumstances, multitasking and parallel processing may be advantageous. Likewise, while several specific implementation details are contained in the above discussions, these should not be construed as limitations on the scope of the subject matter described herein, but rather as descriptions of features that may be specific to particular embodiments. Certain features that are described in this specification in the context of separate embodiments can also be implemented in combination in a single embodiment. Conversely, various features that are described in the context of a single embodiment can also be implemented in multiple embodiments separately or in any suitable sub-combination. Moreover, although features may be described above as acting in certain combinations and even initially claimed as such, one or more features from a claimed combination can in some cases be excised from the combination, and the claimed combination may be directed to a sub-combination or variation of a sub-combination.
[00071] The term “memory,” as used herein relates to a volatile or persistent medium, such as a magnetic disk, or optical disk, in which a computer can store data or software for any duration. Optionally, the memory is non-volatile mass storage such as physical storage media. Furthermore, a single memory may encompass and in a scenario wherein computing system is distributed, the processing, memory and/or storage capability may be distributed as well.
[00072] Throughout the present disclosure, the term ‘server’ relates to a structure and/or module that include programmable and/or non-programmable components configured to store, process and/or share information. Optionally, the server includes any arrangement of physical or virtual computational entities capable of enhancing information to perform various computational tasks.
[00073] Throughout the present disclosure, the term “network” relates to an arrangement of interconnected programmable and/or non-programmable components that are configured to facilitate data communication between one or more electronic devices and/or databases, whether available or known at the time of filing or as later developed. Furthermore, the network may include, but is not limited to, one or more peer-to-peer network, a hybrid peer-to-peer network, local area networks (LANs), radio access networks (RANs), metropolitan area networks (MANS), wide area networks (WANs), all or a portion of a public network such as the global computer network known as the Internet, a private network, a cellular network and any other communication system or systems at one or more locations.
[00074] Throughout the present disclosure, the term “process”* relates to any collection or set of instructions executable by a computer or other digital system so as to configure the computer or the digital system to perform a task that is the intent of the process.
[00075] Throughout the present disclosure, the term ‘Artificial intelligence (AI)’ as used herein relates to any mechanism or computationally intelligent system that combines knowledge, techniques, and methodologies for controlling a bot or other element within a computing environment. Furthermore, the artificial intelligence (AI) is configured to apply knowledge and that can adapt it-self and learn to do better in changing environments. Additionally, employing any computationally intelligent technique, the artificial intelligence (AI) is operable to adapt to unknown or changing environment for better performance. The artificial intelligence (AI) includes fuzzy logic engines, decision-making engines, preset targeting accuracy levels, and/or programmatically intelligent software.

Claims
I/We Claim:
1. A system for extracting protein from human nail samples, comprising: a lysis buffer chamber for dissolving nail samples, wherein the lysis buffer chamber further comprise a lysis buffer solution; a sample transfer mechanism for transferring dissolved nail samples into separate tubes; a filtration unit for separating protein solutions from undissolved sample components through centrifugation; a pulverization apparatus for pulverizing undissolved sample components using a magnetic stirrer; a temperature control unit maintaining samples at specified temperatures ranging from about 40 degree Celcius to about 90 degree Celcius, during processing steps; a supernatant separation mechanism for separating supernatants from undissolved samples by centrifugation; an acetone precipitation unit for precipitating sample fractions; and a sample resuspension chamber for resuspending precipitated sample fractions in a suitable sample buffer.
2. The system of claim 1, wherein the lysis buffer solution further comprises:
from about 0.1M to about 0.2M NaOH;
Sodium Dodecyl Sulphate (SDS) at 1% by weight;
ß-mercaptoethanol (ß ME) at 2% by weight; and
0.01 M Ethylenediaminetetraacetic Acid (EDTA).
3. The system of claim 1, further comprising a data acquisition and control module for monitoring and controlling the sample preparation process, including temperature regulation and timing of sample processing steps.
4. The system of claim 1, further comprising a user interface for inputting sample-specific parameters and controlling the sample preparation process.
5. A method for extracting protein from human nail samples, comprising: dissolving nail samples in a lysis buffer solution; transferring dissolved nail samples into separate tubes by using a sample transfer mechanism; separating protein solutions from undissolved sample components through centrifugation using a filtration unit; pulverizing undissolved sample components using a magnetic stirrer; separating supernatants from undissolved samples using a supernatant separation mechanism; precipitating sample fractions using acetone precipitation; and resuspending precipitated sample fractions in a suitable sample buffer.
6. The method of claim 5, further comprising the step of maintaining samples at specified temperatures ranging from about 40-degree Celsius to about 90-degree Celsius, during processing steps using a temperature control unit.
7. The method of claim 5, wherein sample-specific parameters are inputted and the sample preparation process is controlled using a user interface.
8. The method of claim 5, further comprising the step of resuspending the pelletized proteins in a suitable sample buffer after the acetone precipitation.
9. The method of claim 5, further comprising the step of repeating the pulverization, centrifugation, and supernatant separation steps to maximize the yield of extracted proteins.
10. The method of claim 5, further comprising step of characterizing extracted protein from human nail samples by using characterization techniques selected from Fourier-Transform Infrared (FTIR) spectroscopy, X-ray Diffraction (XRD), Scanning Electron Microscopy - Energy Dispersive X-ray Spectroscopy (SEM-EDX), Inductively Coupled Plasma (ICP) analysis and Thermogravimetric (TG) analysis.

PURIFICATION METHOD FOR KERATIN EXTRACTION FROM HUMAN NAIL WASTES AND DETECTION OF HEAVY METALS
Abstract
The present disclosure presents a system designed for the efficient extraction of proteins from nail samples. The system comprises a lysis buffer chamber housing a specialized lysis buffer solution engineered for dissolving nail samples effectively. Following dissolution, a sample transfer mechanism is employed to transfer the dissolved samples into separate tubes. The system further incorporates a filtration unit, utilizing centrifugation to segregate protein solutions from the undissolved sample components. Any undissolved sample components are then subjected to a pulverization apparatus employing a magnetic stirrer to ensure thorough pulverization. A temperature control unit within the system maintains the samples at specified temperatures, ranging from about 40 degrees Celsius to about 90 degrees Celsius, throughout various processing steps to ensure optimal conditions for protein extraction. The system also encompasses a supernatant separation mechanism, which employs centrifugation to segregate supernatants from undissolved samples. An acetone precipitation unit is integrated for precipitating sample fractions, which are subsequently transferred to a sample resuspension chamber where the precipitated sample fractions are resuspended in a suitable sample buffer for further analysis or utilization. The disclosure provides a comprehensive, streamlined, and effective approach for protein extraction from nail samples, thus paving the way for enhanced utilization of such biological materials in a variety of scientific and industrial applications. , Claims:Claims
I/We Claim:
1. A system for extracting protein from human nail samples, comprising: a lysis buffer chamber for dissolving nail samples, wherein the lysis buffer chamber further comprise a lysis buffer solution; a sample transfer mechanism for transferring dissolved nail samples into separate tubes; a filtration unit for separating protein solutions from undissolved sample components through centrifugation; a pulverization apparatus for pulverizing undissolved sample components using a magnetic stirrer; a temperature control unit maintaining samples at specified temperatures ranging from about 40 degree Celcius to about 90 degree Celcius, during processing steps; a supernatant separation mechanism for separating supernatants from undissolved samples by centrifugation; an acetone precipitation unit for precipitating sample fractions; and a sample resuspension chamber for resuspending precipitated sample fractions in a suitable sample buffer.
2. The system of claim 1, wherein the lysis buffer solution further comprises:
from about 0.1M to about 0.2M NaOH;
Sodium Dodecyl Sulphate (SDS) at 1% by weight;
ß-mercaptoethanol (ß ME) at 2% by weight; and
0.01 M Ethylenediaminetetraacetic Acid (EDTA).
3. The system of claim 1, further comprising a data acquisition and control module for monitoring and controlling the sample preparation process, including temperature regulation and timing of sample processing steps.
4. The system of claim 1, further comprising a user interface for inputting sample-specific parameters and controlling the sample preparation process.
5. A method for extracting protein from human nail samples, comprising: dissolving nail samples in a lysis buffer solution; transferring dissolved nail samples into separate tubes by using a sample transfer mechanism; separating protein solutions from undissolved sample components through centrifugation using a filtration unit; pulverizing undissolved sample components using a magnetic stirrer; separating supernatants from undissolved samples using a supernatant separation mechanism; precipitating sample fractions using acetone precipitation; and resuspending precipitated sample fractions in a suitable sample buffer.
6. The method of claim 5, further comprising the step of maintaining samples at specified temperatures ranging from about 40-degree Celsius to about 90-degree Celsius, during processing steps using a temperature control unit.
7. The method of claim 5, wherein sample-specific parameters are inputted and the sample preparation process is controlled using a user interface.
8. The method of claim 5, further comprising the step of resuspending the pelletized proteins in a suitable sample buffer after the acetone precipitation.
9. The method of claim 5, further comprising the step of repeating the pulverization, centrifugation, and supernatant separation steps to maximize the yield of extracted proteins.
10. The method of claim 5, further comprising step of characterizing extracted protein from human nail samples by using characterization techniques selected from Fourier-Transform Infrared (FTIR) spectroscopy, X-ray Diffraction (XRD), Scanning Electron Microscopy - Energy Dispersive X-ray Spectroscopy (SEM-EDX), Inductively Coupled Plasma (ICP) analysis and Thermogravimetric (TG) analysis.