Rapid sterility testing method and rapid sterility testing platform for safety verification of biopharmaceuticals.

Abstract

The present invention relates to an integrated system and methodology for dramatically reducing the time required for sterility testing for safety verification in the manufacturing process of biopharmaceuticals. Specifically, it provides a rapid sterility testing method and a rapid sterility testing platform for safety verification of biopharmaceuticals, comprising electromagnetic nanobeads coated with proteins that can bind to microorganisms constituting the system, a biochip specialized for nanoparticle analysis, automated imaging equipment and analytical software, and a process and method for performing rapid sterility verification.

Description

Technical Field

[0001] The present invention relates to a rapid sterility test method and a rapid sterility test platform for verifying the safety of biopharmaceuticals. More specifically, it relates to an integrated system and its methodology that epochally shorten the required time for sterility verification tests in the manufacturing process of biopharmaceuticals. Specifically, it includes magnetic particles coated with proteins that can bind to microorganisms constituting the system, a biochip specialized for microorganism analysis combined with nanoparticles, automated imaging equipment and analysis software, and the process and method for performing rapid sterility verification. The present invention relates to a rapid sterility test method and a rapid sterility test platform for verifying the safety of biopharmaceuticals.

Background Art

[0002] Sterility testing is a test method for quality control of pharmaceuticals, detecting bacteria and fungi. It is applied to raw materials and formulations during the manufacturing of sterile pharmaceuticals. A specified amount of sample is taken, and the experiment is conducted using a specified culture medium and method to check for the presence of bacteria and fungi determined to be originating from the sample. Currently, there are two types of sterility testing for pharmaceuticals: the membrane filter method and the direct method, with the membrane filter method being the standard. The membrane filter method involves passing the biopharmaceutical through a filter to purify non-microbial components, concentrating the sample, and then placing it in a microbial growth medium. The turbidity, which increases as microorganisms divide, is then detected. This method typically requires a long and labor-intensive process of two weeks. However, currently, the majority of biopharmaceuticals, unlike conventional chemically synthesized drugs, have a much shorter shelf life than two weeks. This creates a problem where sterility verification test results are derived after the drug has been administered to patients immediately after manufacturing. This poses a clinical risk of potentially causing microbial infections in patients. Furthermore, because biopharmaceuticals, including cell therapies, are manufactured on a patient-by-patient basis, standardized management is difficult. Although management regulations for these advanced pharmaceuticals have only recently been established, rapid bacterial / fungal testing methods suitable for sterility testing of cell therapies have not yet been developed. As a result, microbiological safety management remains a limiting factor for both producers and patients. Therefore, with the increasing use of biopharmaceuticals, there is a growing need for the development of rapid sterility verification tests that address the shortcomings of conventional sterility testing methods, which require long detection times.

[0003] To address this issue and shorten the time required for sterility verification testing, methods have been proposed that eliminate the culture stage and detect microorganisms directly. Representative techniques include solid-phase cytometry and polymerase chain reaction (PCR). In solid-phase cytometry, the biopharmaceutical is passed through a membrane filter, and the microorganisms captured on the filter are stained and detected under a microscope. In PCR, detection is achieved by utilizing the fact that the signal is amplified when microorganisms are present via primers specific to the microorganism to be detected. Both techniques have the advantage of being able to detect microorganisms within three hours, but neither method is universally applicable to a wide variety of microorganisms and types of biopharmaceuticals, and both have a high rate of false positives or false negatives, making it difficult to replace conventional sterility verification testing of biopharmaceuticals. Therefore, current technological development is focused on methods for sensing metabolites such as carbon dioxide or adenosine triphosphate (ATP) produced by microbial growth. However, even with such techniques, the limitations of the purification efficiency of the membrane filter method prevent the effective separation of components and microorganisms within biopharmaceuticals. This leads to interference and false-positive problems caused by metabolites produced by cells other than microorganisms, requiring a long time of about 5 days for meaningful signal measurement. To solve the above problems, this invention presents a methodology and integrated system that separates microorganisms from biopharmaceuticals containing cells and microorganisms with superior purification and concentration efficiency compared to the membrane filter method, and enables the specific and rapid detection of microbial metabolites. [Overview of the project] [Problems that the invention aims to solve]

[0004] To solve the aforementioned problems, the present invention provides an integrated system and methodology that dramatically reduces the time required for sterility testing for safety verification in the manufacturing process of biopharmaceuticals. Specifically, it aims to provide a rapid sterility testing method and a rapid sterility testing platform for biopharmaceutical safety verification, which include magnetic particles coated with proteins that can bind to microorganisms constituting the system, a biochip specialized for microbial analysis bound to nanoparticles, automated imaging equipment and analytical software, and a process and method for performing rapid sterility verification. [Means for solving the problem]

[0005] To solve the aforementioned problems, the present invention provides a rapid sterility testing method using a biochip comprising one or more chambers into which a sample and a control group are loaded, and one or more magnets installed on one side of each chamber, comprising the steps of: mixing magnetic particles with a substance to be tested to bind the analyte to the magnetic particles; separating the magnetic particles from the substance to be tested to produce a sample; and injecting the separated sample and indicator substance into the chambers of the biochip and culturing them to determine the presence of the analyte, wherein the step of determining the presence of the analyte involves comparing the fluorescence or hue of the sample and control group images after culturing to determine the presence or absence of the analyte inside the sample.

[0006] In one embodiment, the substance to be tested may be a biopharmaceutical, a cell therapy agent, a clinical sample, a chemical drug, a cosmetic, a food product, or an environmental sample.

[0007] In one embodiment, the analyte may be bacteria, mycoplasma, or fungi.

[0008] In one embodiment, the step of preparing the sample may include the step of concentrating the analyte bound to the magnetic particles on a growth medium.

[0009] In one embodiment, the growth medium may include tryptic soy broth, Muller-Hinton broth, fluid thioglycolate medium, Sabouraud broth, and Luria-Bertani broth.

[0010] In one embodiment, the step of determining the presence of the analyte may include the steps of: separating the chamber from the magnet; injecting an indicator substance, a sample, and a control group into the chamber and culturing them for a certain period of time; bringing the chamber closer to the magnet after the culturing is complete; and, after photographing the inside of the chamber to obtain an image, comparing the images taken from the separated sample and the control group.

[0011] In one embodiment, the analyte combines with the magnetic particles to form a composite, and the composite can be moved to one side of the chamber by the magnet when the chamber is brought closer to the magnet.

[0012] In one embodiment, the culture may be carried out for 4 to 48 hours.

[0013] In one embodiment, the image acquisition can be performed at intervals of 10 to 120 minutes.

[0014] In one embodiment, the indicator substance may fluoresce or change color depending on the analyte.

[0015] In one embodiment, the indicator substance may be a redox indicator, a pH indicator, a dye-based reagent, an enzyme reaction indicator, or an antibody-based reagent.

[0016] The present invention also provides a biochip for a rapid sterile testing platform, comprising a lower plate including a chamber into which a control group and samples are loaded, and an upper plate fitted to the lower plate and including a magnet, wherein the lower plate includes a bottom surface, a central axis protruding upward from the bottom surface, and one or more chambers positioned at a certain distance from the central axis, and the upper plate includes a central axis insertion port into which the central axis of the lower plate is inserted, a chamber insertion port into which the chambers are inserted, and a magnet positioned on one side of the chamber insertion port.

[0017] In one embodiment, the chamber may include one or more sample chambers and one or more control chambers.

[0018] In one embodiment, the chambers may be installed at equal intervals at uniform distances from the central axis.

[0019] In one embodiment, the chamber may further contain magnetic particles and an indicator material.

[0020] In one embodiment, the magnetic particles may combine with the analyte to form a composite.

[0021] In one embodiment, the magnetic particles may contain functional groups or substances on their surface that can bind to microorganisms.

[0022] In one embodiment, the composite may be positioned on one side of the chamber using the magnet during fluorescence or optical observation using the indicator material.

[0023] In one embodiment, the upper plate may rotate at a constant angle about the central axis.

[0024] In one embodiment, the control group chamber insertion portion may have an arc shape centered on the central axis.

[0025] In one embodiment, the magnet may be one whose distance from the chamber changes according to the rotation of the upper plate.

[0026] In one embodiment, rotating means capable of rotating the upper plate may be installed on the outer surface of the upper plate.

[0027] The present invention also provides a rapid sterility test platform including the biochip for the rapid sterility test platform.

[0028] In one embodiment, the rapid sterility test platform may include the biochip, a rotational power unit that rotates the upper plate of the biochip, an illumination unit installed above or below the biochip, and an image acquisition unit that photographs the inside of the chamber of the biochip.

[0029] In one embodiment, the image acquisition unit can acquire a fluorescence image or an optical image inside the chamber.

[0030] In one embodiment, the rapid sterility test platform can compare the fluorescence or hue of the acquired first image and the current image to determine the presence or absence of microorganisms inside the sample.

Advantages of the Invention

[0031] The rapid sterility verification method proposed in the present invention can shorten the sterility verification test that conventionally took 14 days to within one day (24 hours), and can quickly ensure the safety of biopharmaceuticals. Considering that biopharmaceuticals produced using human-derived substances such as cells and proteins have a short expiration date and must be quickly administered to patients, shortening the time required for the overall test is beneficial for maintaining the quality of biopharmaceuticals at the time of patient administration, and at the same time can improve the therapeutic effect on patients through rapid administration. Also, from a social and corporate perspective, since the safety verification process is shortened by more than 13 days, it is possible to save a huge amount of logistics costs.

[0032] Conventional sterility verification tests require continuous monitoring for contamination at regular intervals, such as 24, 48, and 72 hours after microbial culture, for up to 14 days. In contrast, the rapid sterility verification method of the present invention performs continuous imaging on the system in real time or at approximately 30-minute intervals to determine the presence or absence of contamination. From a temporal perspective, it offers exceptionally high resolution and allows for more accurate analysis based on fluorescence intensity values. Furthermore, since it does not require direct observation at regular intervals via an automated system, it minimizes the labor and human resources required in practical applications compared to conventional methods.

[0033] From a technical standpoint, recent technologies, including conventional sterility verification methods, utilize membrane filtration to concentrate contaminating microorganisms in the biopharmaceutical sample to be analyzed. For sterility verification, it is necessary to be able to universally detect microorganisms ranging from 1 μm in size to 10 μm in size, but with filtration methods, microbial sample loss can occur depending on the pore size. When using filters with small pore sizes, substances such as cells and proteins present in the biopharmaceutical are also concentrated, reducing the sensitivity of observation of microorganisms or their metabolites in further analysis. This invention uses functionalized nanoparticles that can specifically bind to microorganisms from substances present in the biopharmaceutical, enabling the isolation and concentration of microorganisms regardless of their size. Compared to conventional filtration methods, this allows for more sensitive analysis in detecting microbial concentration. Therefore, the concentration of microorganisms in a smaller volume accelerates the reaction of the laserzlin stain, enabling faster measurement of metabolites compared to prior art. In particular, in sterility verification tests, viability testing, which detects metabolites (typically ATP, CO2, and NADH) to target all microbial species, is recommended as the standard test. However, substances other than microorganisms, such as cells, contained within biopharmaceuticals also produce metabolites. Therefore, utilizing methods for isolating and concentrating such microorganisms has the advantage of greatly improving the specificity of the analysis.

[0034] Nanoparticle-based methods for isolating and concentrating microorganisms can be applied not only to laserzlin stains but also to analyses utilizing molecular diagnostic methods such as polymerase chain reaction (PCR), next-generation sequencing methods, and advanced fluid analysis instruments. The characteristics of these technologies offer flexibility in their application. For example, in the case of biopharmaceutical samples where contamination has been confirmed, molecular diagnostic methods can be used to identify microbial species and track the contamination pathway, or to prevent secondary contamination. Alternatively, they can be applied to samples requiring sterility verification, such as food, cosmetics, and environmental monitoring, and are used according to the purpose.

[0035] The invention of a chip specifically designed for nanoparticle-based analytical methods prevents growth inhibition of microorganisms bound to nanoparticles, separated, and concentrated, enabling stable observation of microbial signals. While normal growth is not possible if microorganisms bound to nanoparticles persistently aggregate against the wall of the biochip, the biochip included in this invention allows for the application or removal of magnetism with simple operation, enabling normal microbial growth. Therefore, compared to not using the chip, the production of microbial metabolites increases, and signals to microorganisms or their metabolites can be observed more rapidly and sensitively.

[0036] Continuous fluorescence imaging is performed using the miniaturized 224mm × 120mm × 140mm imaging system proposed in this invention. The imaging system includes an xyz motor-driven stage with adjustable imaging area and focus for chip samples, a light source for fluorescence imaging, a 4X objective lens, and a CCD camera, and is manufactured in a simplified form specifically for the purpose of rapid sterility verification. This is significantly cheaper than conventional commercially available plate reader equipment, can be widely adopted in industry in a simplified form, can be easily placed in an incubator, and is advantageous for temperature maintenance and space optimization. [Brief explanation of the drawing]

[0037] [Figure 1] This figure shows the configuration and operating principle of a biochip for a rapid sterility testing platform according to one embodiment of the present invention. [Figure 2] This figure shows the shape of a biochip for a rapid sterility testing platform according to one embodiment of the present invention, where (a) shows the overall shape, (b) shows the position of the chamber when rotating in reverse, and (c) shows the position of the chamber when rotating in forward. [Figure 3] This figure shows the configuration of the upper plate of a biochip for a rapid sterility testing platform according to one embodiment of the present invention. [Figure 4] This figure shows a rapid sterility testing platform according to one embodiment of the present invention. [Figure 5] This figure shows a rapid sterility testing method using a rapid sterility testing platform according to one embodiment of the present invention. [Figure 6] This figure shows a concentration method using magnetic nanoparticles according to one embodiment of the present invention. [Figure 7] This figure shows the results of microbial growth with and without magnetism according to one embodiment of the present invention. [Figure 8] This figure compares a rapid sterility test method according to one embodiment of the present invention with a conventional sterility verification method. [Figure 9] This figure shows the results of rapid sterility testing on various biopharmaceuticals using one embodiment of the present invention. [Figure 10] This figure shows the presence or absence of microbial contamination as observed visually using one embodiment of the present invention. [Modes for carrying out the invention]

[0038] Preferred embodiments of the present invention will be described in detail below. When describing the present invention, if it is determined that a specific description of related prior art may obscure the gist of the invention, such detailed description will be omitted.

[0039] Throughout the specification, singular expressions should be understood to include plural expressions unless otherwise clearly stated in the portal tract, and terms such as “includes” or “having” should be understood to indicate the existence of the described features, numbers, stages, operations, components, parts, or combinations thereof, without pre-existing exclusion of the existence or possibility of adding one or more other features, numbers, stages, operations, components, parts, or combinations thereof. Furthermore, when carrying out a method or manufacturing method, each step comprising the method may be carried out in a different order than specified unless otherwise clearly stated in the portal tract, that is, each step may be carried out in the same order as specified, substantially simultaneously, or in reverse order.

[0040] The technology disclosed herein is not limited to the examples described herein and can be embodied in other forms. The examples presented herein are provided solely to ensure that the disclosed content is thorough and complete, and that the technical idea of ​​the technology can be fully conveyed to those skilled in the art. In the drawings, the dimensions of the components, such as width and thickness, have been slightly enlarged to clearly represent the components of each device. The drawings are generally described from the observer's perspective, and when it is mentioned that one element is located on top of another, this includes the meaning that the one element is located directly on top of the other, or that further elements may be interposed between them. Furthermore, those with ordinary skill in the art can realize the idea of ​​the present invention in various other forms without departing from the technical idea of ​​the present invention. Note that the same reference numerals in multiple drawings refer to substantially the same elements.

[0041] In this specification, the term "and / or" includes any combination of the listed items or any one of the listed items. In this specification, "A or B" may include "A", "B", or "both A and B".

[0042] The present invention relates to a rapid sterility testing method using a biochip comprising one or more chambers into which a sample and a control group are loaded, and one or more magnets installed on one side of each chamber, comprising the steps of: mixing magnetic particles with a substance to be tested to bind the analyte to the magnetic particles; separating the magnetic particles from the substance to be tested; and injecting the separated magnetic particles and indicator substance into the chambers of the biochip and culturing them to determine the presence of the analyte, wherein the step of determining the presence of the analyte involves comparing the fluorescence or hue of the sample and control group images after culturing to determine whether or not the analyte is present inside the sample.

[0043] In the case of biological preparations such as biopharmaceuticals, the presence of undesirable microorganisms can significantly reduce their safety. This invention provides a rapid sterility test method that can quickly confirm the presence or absence of microorganisms compared to conventional methods, in order to verify the safety of such pharmaceuticals.

[0044] The biochip of the present invention may include one or more chambers into which a sample and a control group are loaded, and one or more magnets installed on one side of each chamber. The structure of such a biochip will be described later.

[0045] Magnetic particles can be mixed with the substance to be tested to bind the analyte to the magnetic particles. The magnetic particles refer to particles that bind to the analyte present in the substance to be tested, and may contain paramagnetic or ferromagnetic materials. In this invention, the analyte can be separated from the substance to be tested, and the magnetic particles can be easily moved using magnetism. Furthermore, the analyte can bind to the magnetic particles to form a composite. As described above, the composite means that the composite is bound to the surface of the magnetic particles, and as will be described later, the surface of the magnetic particles contains functional groups or substances that can bind to microorganisms, so that it can selectively bind to the analyte to form a composite.

[0046] In this case, the magnetic particles can be any magnetic particles that have been conventionally used for microbial separation, and can be manufactured in the form of microparticles or microrods. Furthermore, the magnetic particles may be nanoparticles having a nanoscale.

[0047] The magnetic particles may contain functional groups or substances on their surface that can bind to microorganisms. This allows for selective binding to analytes present in the substance being tested. In this case, the functional groups or substances may be beta-2-glycoprotein 1, mannose-binding lectin, antibodies, aptamers, antibiotics, or polymers.

[0048] In this context, the substance to be tested refers to a substance that requires ensuring stability against microorganisms, and specifically may be a biopharmaceutical, cell therapy agent, clinical sample, chemical drug, cosmetic, food, or environmental sample. In particular, the biopharmaceutical may be manufactured based on proteins, peptides, antibodies, or vaccines, or based on genes, stem cells, human-derived cells, human cells, or tissues, and the same applies to the clinical sample and cell therapy agent. In the case of such substances to be tested, ensuring safety as described above is essential because they can have a fatal effect on the human body if contaminated with microorganisms.

[0049] The analyte refers to microorganisms that reduce the safety of the substance being tested, particularly pharmaceuticals, and may include bacteria, mycoplasmas, and fungi. When such microorganisms are found in the pharmaceutical, it is generally said that the pharmaceutical is contaminated, and due to the nature of the pharmaceutical, if its safety is reduced due to such contamination, not only a single product but also the entire batch, products produced on the same day, or products from the same production line should be discarded.

[0050] As described above, after the analyte has bound to the magnetic particles, the magnetic particles can be separated from the substance to be tested. In this case, since the analyte is bound to the magnetic particles, the separation can be performed simply by washing. Furthermore, after washing is complete, a magnet can be used to remove the washing solution and foreign matter. As described above, in the present invention, since the analyte is separated using magnetic particles, when a magnet is used, the magnetic particles and the analyte bound to them can be easily moved. That is, after washing is complete, the magnetic particles can be separated by inserting a magnetic rod into the washing container, or separately, by bringing the magnet close to the side or bottom of the washing container and then discharging the washing solution and foreign matter from inside, separation can be performed without loss of the magnetic particles. As a result, even when the concentration of the analyte is very low, it can be separated together with the magnetic particles, which can greatly reduce the probability of false negatives.

[0051] Furthermore, it is preferable to concentrate the analyte bound to the magnetic particles on a growth medium. Concentration on the growth medium can be performed by fixing the magnetic particles to the side of the container using a magnet, removing the analyte from the supernatant, and then resuspending the growth medium. An appropriate amount of growth medium can be used depending on the size of the container and the amount of magnetic particles; specifically, it is preferable to resuspend using approximately 100 microliters of growth medium. In this case, it is preferable to wash with the growth medium three or more times in order to effectively recover the magnetic particles remaining on the side of the container. The growth medium plays a role in promoting the growth of the analyte and reducing the detection time, and concentration on a small amount of growth medium can increase the concentration of the analyte and further shorten the detection time.

[0052] Furthermore, the growth medium can be any medium capable of culturing the analyte without limitation, but preferably it may include tryptic soy broth, Muller-Hinton broth, fluid thioglycolate medium, Sabouraud broth, or Luria-Bertani broth.

[0053] The separated magnetic particles and indicator material can be injected into a biochip chamber and cultured to determine the presence of the analyte. The biochip used may include one or more chambers into which the sample and control are loaded, and one or more magnets installed on one side of each chamber, which will be described later.

[0054] The step of determining the presence of the analyte may include the steps of: moving the chamber away from the magnet; injecting an indicator substance, a sample, and a control group into the chamber and culturing them for a certain period of time; after the culturing is complete, moving the chamber closer to the magnet; and after photographing the inside of the chamber to obtain an image, comparing the images taken from the sample and the control group.

[0055] The chamber and the magnet can be separated. As described above, when the analyte attached to the magnetic particles is cultured, it is preferable that the composite formed by the binding of the magnetic particles and the analyte floats within the chamber. Therefore, by separating the chamber and the magnet, the magnetic attraction can be reduced, and the composite can float freely from inside the chamber.

[0056] The indicator substance, sample, and control group can be injected into the chamber and cultured for a certain period of time. In the case of the sample, as described above, the sterility test may be performed by separating it from the desired substance to be tested, or it may be a composite in which the analyte is bound to the surface of the magnetic particles.

[0057] The control group refers to a substance whose safety has been verified, i.e., a negative control group, and preferably means a sample that is identical to the sample and has the same components as the analyte excluding the analyte. In other words, the control group may include magnetic particles that do not form a complex and the same growth medium as the sample. After culturing such a control group and the sample simultaneously, the presence or absence of the analyte in the chamber can be confirmed by observing the changes in the chamber.

[0058] Furthermore, as described above, the sample can be manufactured and used separately from the biochip, but it can also be concentrated and used inside the biochip. In this case, the biochip contains the magnetic particles inside, and after supplying the substance to be tested into the biochip, a cleaning process can be performed using the magnet installed in the biochip. As described above, during such a cleaning process inside the biochip, the composite can be collected on one side of the chamber by the magnet, and then the cleaning solution and foreign matter can be removed.

[0059] In other words, as described above, when the sample is prepared outside the biochip, a separate chamber or container can be used, and when the sample is prepared using the biochip, the sample can be prepared using the internal chamber of the biochip.

[0060] The indicator substance is used to confirm the presence of an analyte that forms a complex with the sample during the culture of the sample in the chamber, and may be a redox indicator, a pH indicator, a dye-based reagent, an enzyme reaction indicator, or an antibody-based reagent. Such an indicator substance can change its hue or fluorescence in response to chemical changes in the chamber due to microbial respiration, digestion, or reproduction, thereby confirming the presence or absence of microorganisms in the sample chamber.

[0061] In this case, the indicator substance can be injected before or after the injection of the sample and control group, or it can be supplied loaded inside the biochip. In this case, user convenience can be greatly improved, but the biochip containing the indicator substance must be replaced depending on the type of sample, which can increase the cost of analysis. Therefore, it is preferable to appropriately adjust the injection timing of the indicator substance depending on the user's situation and surrounding environment.

[0062] The aforementioned redox indicator refers to a substance whose hue or fluorescence changes due to oxidation or reduction reactions, and is preferably laserzrin. Generally, all cells, including microorganisms, carry out a variety of oxidation and reduction reactions internally. In this case, if such a redox indicator is present, the hue or fluorescence can change due to these redox reactions. In particular, since laserzrin is cell-permeable, such changes in fluorescence or hue can be exhibited even more clearly when redox reactions occur inside the cell.

[0063] The aforementioned Lezazrin is a type of phenoxazine dye that is non-toxic, cell-permeable, and redox-sensitive. When it reacts with living cells, it visibly changes from blue to red and simultaneously exhibits fluorescence, which can be measured through optical or fluorescence observation. In other words, the appearance of such a color change confirms the presence of living cells within the sample, and the absence of fluorescence or color change after culturing means that sterility can be verified.

[0064] As described above, after injecting the sample and control group into the chamber, they can be cultured for a certain period of time. In this case, it is preferable that the culture is carried out for 4 to 48 hours. Conventional sterility verification tests use turbidity caused by the microorganisms themselves, so long-term culture (2 weeks) is required. However, in the present invention, the presence or absence of microorganisms is sensed using the color change of the oxidation-reduction indicator, so the presence or absence of microorganisms can be confirmed even with a short culture period as described above. If the culture is carried out for less than 4 hours, it may be difficult to observe changes because the culture time is very short, and if it exceeds 48 hours, the overall test time increases, which is inefficient.

[0065] After the culture is completed as described above, the chamber can be brought close to the magnet. During this process, the magnetic particles in the control chamber and the composite (magnetic particles + microorganisms) in the sample chamber can be collected on one side of the chamber by the magnet.

[0066] When the above-described collection occurs, the magnetic particles and microorganisms inside the chamber are located off-center, making it possible to obtain a clearer image during the image acquisition process described later.

[0067] After capturing an image of the inside of the chamber, the images captured from the sample and the control group can be compared. As described above, in the case of the control group, sterility has been verified, so only changes due to the external environment may occur. Therefore, if the changes in the sample are the same as those in the control group, it can be determined that no microorganisms are present in the sample. If the hue or fluorescence of the sample changes, it can be determined that microorganisms are present.

[0068] In this case, it is preferable that the image acquisition is performed at intervals of 10 to 120 minutes. In conventional methods, since the culture period is two weeks, it was common to acquire images on a daily (24-hour) basis. However, in the present invention, the culture time is very short (4 to 48 hours), and if there is a change in hue, it can be immediately determined that microorganisms are present, so images can be acquired at intervals of 10 to 120 minutes, preferably 40 to 80 minutes, and most preferably 60 minutes.

[0069] The present invention also provides a biochip for a rapid sterile testing platform, comprising a lower plate including a chamber into which a control group and samples are loaded, and an upper plate fitted to the lower plate and including a magnet, wherein the lower plate includes a bottom surface, a central axis protruding upward from the bottom surface, and one or more chambers positioned at a certain distance from the central axis, and the upper plate includes a central axis insertion port into which the central axis of the lower plate is inserted, a chamber insertion port into which the chambers are inserted, and a magnet positioned on one side of the chamber insertion port.

[0070] The lower plate is on which a chamber is installed, and the control group and the sample can be loaded into the chamber.

[0071] The bottom surface 110 of the lower plate is generally flat, but it is preferable that it be manufactured in various shapes depending on the position of the chamber and the shape of the rapid sterile testing platform. In the present invention, it is described based on the assumption that it is manufactured in a circular shape, but it is also possible to manufacture it in square, elliptical, triangular, and polygonal plate shapes, and it is also possible to manufacture it in a plate shape that is not a general plate shape but has a bend or a certain shape at the top or bottom. In addition to a plate shape, it is also possible to manufacture it in a columnar shape or with an inclination in one or various directions.

[0072] In the present invention, it is preferable that the bottom surface 110 be manufactured in the shape of a rectangular plate, as described above. As will be described later, such a rectangular plate shape can be prevented from rotating together with the rotation of the upper plate, even when simply inserted into a groove corresponding to the shape of the bottom surface 110, without the need to install a separate fixing device when it is placed on the rapid sterile testing platform, as described later.

[0073] A central axis 120 and a chamber may be installed on the lower plate.

[0074] The central axis 120 is the part that the upper plate, described later, fits into and rotates at, and can protrude upward from the lower plate. In this case, the central axis 120 may be manufactured to have a circular cross-section so that the upper plate rotates smoothly.

[0075] Furthermore, while it is preferable that the central axis 120 be installed in the center of the bottom surface 110, it may also be installed at a position a certain distance away from the center of the bottom plate for the purpose of fixing the bottom plate or installing the chamber.

[0076] Furthermore, the central shaft 120 can be manufactured simply in a cylindrical shape, but a separation prevention means may be installed at the end that is inserted into the upper plate. The separation prevention means is a part that prevents the upper plate from coming off after the upper plate has been fitted, and is manufactured to be larger than the diameter of the central shaft 120 insertion opening installed in the upper plate, thereby preventing the upper plate from coming off after being coupled with the upper plate. In addition, to facilitate the coupling of the central shaft 120 and the central shaft insertion opening 210, the central shaft 120 can be manufactured in a pipe shape with a hole in the center, and a slit may be installed in a part of the upper part, so that the diameter decreases when passing through the central shaft insertion opening 210.

[0077] The chamber is the portion into which the sample and control group are loaded, and may be manufactured in a form that is closed at the bottom and open at the top. It is also preferable that it be manufactured in a pipe shape with an open interior for the loading described above, and that the bottom be made optically transparent to facilitate image acquisition, as described later. Furthermore, the chamber may be manufactured with a circular, elliptical, square, or polygonal cross-section, but may be manufactured to be circular for mixing efficiency and culture uniformity. In other words, it is most preferable that the chamber be manufactured in a circular pipe shape with a closed bottom and an open top.

[0078] Furthermore, the upper end of the chamber may be manufactured in an open form, but preferably a polymer film may be installed. The polymer film serves to prevent contact with external contaminants after the sample and control group have been loaded, and for the convenience of loading the sample and control group, a "-" or "+" shaped notch may be provided at the top. The sample and control group are generally loaded using a dropper, pipette, or syringe. Therefore, when loading the sample and control group, they may be loaded through the notch, and after loading is complete, the notch is closed elastically so that contact with external contaminants, especially microorganisms, can be blocked.

[0079] The chamber may include one or more sample chambers 131 and one or more control chambers 132. In the case of the sample chamber 131, it is the chamber into which the sample to be tested for sterility is loaded, and in the case of the control chamber 132, it is the chamber into which a solution whose sterility has been verified, i.e., the control group, is loaded (see Figure 1(a)). By loading the sample and the control group simultaneously as described above, the sample and the control group can be compared under the same conditions, and this makes it easy to identify if the sample is contaminated.

[0080] Furthermore, while one sample chamber 131 and one control chamber 132 may be installed, it is also possible to install multiple sample chambers. When multiple sample chambers 131 are installed, sterility testing can be performed on multiple samples simultaneously, and when multiple control chambers 132 are installed, sterility testing can be performed under a variety of conditions.

[0081] The chambers may be installed at equal intervals at uniform distances from the central axis 120. In the case of the chambers, as will be described later, the top plate rotates after being inserted into the chamber inlet. Therefore, it is preferable that the chambers be arranged at a constant distance from the central axis 120 to facilitate such rotation of the top plate. Furthermore, it is preferable that the chambers be installed at equal intervals to maintain the same distance between each chamber and the magnet 230, which will be described later. If the intervals between the chambers are not uniform, the distances from the magnet 230 will be different for each chamber, so the magnetic particles inside each chamber will receive different magnetism, which means that the experimental results for each chamber may differ.

[0082] The chamber may further contain magnetic particles and an oxidation-reduction indicator.

[0083] The magnetic particles may bind with microorganisms to form a complex (see Figure 1(a)). As the magnet 230 approaches one side of the chamber during the forward rotation process of the upper plate described later, these magnetic particles can move magnetically to the wall on one side of the chamber (Figure 1(b)). This allows the central part of the chamber to be free of the microorganisms (complex) bound to the magnetic particles, and a clearer image can be obtained. In other words, the complex may be located on one side of the chamber using the magnet 230 during fluorescence or optical observation using the indicator substance described later.

[0084] The microorganisms can be bound to the magnetic particles using known methods, but preferably, a substance that selectively binds to proteins present on the surface of the microorganisms can be attached to the surface of the magnetic particles before use. In this case, the substance that selectively binds to proteins present on the surface of the microorganisms can be selected appropriately depending on the type of microorganism, but preferably it may be apolipoprotein H, beta-2-glycoprotein 1, mannose-binding lectin, antibody, aptamer, antibiotic, or polymer.

[0085] Furthermore, while the magnetic particles can be used by directly mixing them with the sample as described above, they can also undergo a concentration process using the magnetic particles (see Figure 6). Such a concentration process can be carried out inside the chamber, or in a separate chamber. That is, after mixing the magnetic particles with the substance to be tested, the magnetic particles can be separated, and the microorganisms (analytes) attached to the magnetic particles can also be separated. Using this method, microorganisms can be isolated from the substance to be tested at high concentrations, and these can then be cultured to perform sterility verification.

[0086] Since the indicator substance is the same as the one described above, its explanation will be omitted.

[0087] The upper plate is the part that fits with the lower plate, and as described above, the central shaft 120 formed in the lower plate can be inserted into the central shaft insertion port 210 installed in the upper plate and coupled. That is, the upper plate can be coupled in such a way that it can rotate with respect to the central shaft 120 (Figures 2(b) and (c)). Since the upper plate has a magnet 230 installed on it, which will be described later, the distance between the magnet 230 and the chamber of the lower plate can change in accordance with the rotation described above. In this case, when the magnet 230 moves away, the magnetic nanoparticles inside the chamber can float freely inside the chamber, and when the magnet 230 moves closer, they can be collected on one side of the chamber by the magnet 230 (see Figure 1(b)). This will be described later.

[0088] The upper plate may include a central shaft insertion port 210 into which the central shaft 120 of the lower plate is inserted, a chamber insertion port into which the chamber is inserted, and a magnet 230 installed on one side of the chamber insertion port (see Figure 3).

[0089] The central shaft insertion port 210 is the portion into which the central shaft 120, which is installed on the lower plate, is inserted. It is manufactured in a circular shape corresponding to the central shaft 120, allowing the central shaft 120 to rotate easily. In this case, it is preferable that the central shaft insertion port 210 be located in the center of the upper plate, but if a large number of chambers are installed and eccentric rotation is required, it may be located on one side of the upper plate instead of in the center.

[0090] Furthermore, while the upper plate is preferably manufactured in a circular or elliptical shape as shown in Figure 3, it can also be manufactured in a square or polygonal shape for the arrangement of internal magnets and chambers or for connection with external systems.

[0091] Furthermore, if a separation prevention mechanism is installed on the central axis 120, the separation of the upper and lower plates can be prevented by manufacturing it in a corresponding manner.

[0092] The chamber insertion portion may have an arc shape centered on the central axis 120. As described above, the nanoparticle-based biochip for rapid sterility testing platform of the present invention allows the position of the magnet 230 to be changed by the rotation of the upper plate. In this case, if the chamber insertion portion has the same shape as the chamber, the upper plate cannot rotate, and even if it is simply formed in a straight line, the rotation of the upper plate may be impossible. Therefore, as described above, if it has an arc shape, the chamber can move along the arc shape, and as a result, the upper plate can rotate smoothly.

[0093] In this case, it is preferable that one chamber insertion section is installed for each chamber. That is, in the present invention, since the sample chamber 131 and the control group chamber 132 are installed on the lower plate, a sample chamber insertion port 221 and a control group chamber insertion port 222 can be formed at positions corresponding to each chamber (see Figure 2(a)).

[0094] Furthermore, in the present invention, as will be described later, since magnetic nanoparticles inside the chamber are moved using magnets 230, it is preferable that each chamber has its own magnet 230 and chamber inlet. If multiple chambers share the same chamber inlet, the positions of the magnets 230 corresponding to each chamber may not be uniform, and they may be affected differently by magnetism, which can lead to differences in observation between each chamber.

[0095] A magnet 230 may be installed on one side of the chamber inlet. The magnet 230 is used to collect magnetic nanoparticles inside the chamber to one side of the chamber, and it is preferable that it be installed on one side of the chamber inlet.

[0096] In other words, as the upper plate rotates, the distance between the chamber and the magnet changes, and when the chamber is located away from the magnet 230, the magnetic influence is reduced, allowing the magnetic nanoparticles to float within the chamber. Also, when the chamber is located close to the magnet 230, the magnetic nanoparticles may be collected on one side of the chamber by magnetism (see Figure 1).

[0097] Furthermore, in the case of the magnet, the distance to the chamber can change according to the rotation of the upper plate, but the distance to the chamber can also be increased by the movement of the upper plate. That is, if the chamber insertion opening is formed in a straight line, the upper plate moves linearly rather than rotationally, and in this case, the distance between the magnet and the chamber can also be changed by the linear movement of the upper plate as described above. In this case, it is preferable that the upper plate be manufactured in a polyhedral shape rather than a circular shape to facilitate such linear movement.

[0098] The collection of such magnetic particles can have two effects. First, it can increase the ease and accuracy of observation. As described above, the magnetic particles can bind with microorganisms to form a complex, and in this case, the magnet 230 can collect the complex on one side of the chamber. As a result, after the collection is completed as described above, the concentration of microorganisms in the center of the chamber can be greatly reduced. Generally, when magnetic particles are used, a large amount of magnetic particles float in the chamber, so light does not pass through, and it is difficult to observe the change in hue or fluorescence caused by the indicator substance. Therefore, in conventional methods, after culturing for a long period of time, the magnetic particles (complex) are removed and the change in turbidity or hue is observed. However, in the present invention, as described above, by collecting the complex on one side of the chamber using the magnet, it is possible to obtain a clear image even if only a slight change in hue or fluorescence appears.

[0099] The second effect is that it can promote the growth of microorganisms. Generally, once a certain concentration of microorganisms is reached, the number of individuals does not increase further. This is due to substrate competition, nutrient deficiency, and external contamination, as mentioned above, and for the sustained growth of microorganisms, the concentration needs to be adjusted to below a certain level. In the present invention, by applying this, when the upper plate is rotated forward, the distance between the chamber and the magnet 230 becomes smaller, the concentration of the complex in the center of the chamber decreases, making image acquisition easier. When the upper plate is rotated backward, the distance to the magnet 230 increases, eliminating localized increases in the concentration of microorganisms and increasing the overall culture rate. Therefore, after image acquisition is complete or when rapid culture is required, the upper plate can be rotated backward to increase the distance between the magnet 230 and the chamber. When image acquisition is performed, or when culture is complete and further culture is not required, the upper plate can be rotated forward to bring the distance between the chamber and the magnet closer.

[0100] Therefore, even when the microbial concentration is very low, a high microbial concentration can be obtained by repeating the image acquisition and cultivation process described above many times. This also enables a more rapid and accurate verification of sterility compared to conventional methods.

[0101] The magnet 230 can be installed on one side of the chamber inlet. In particular, it is preferable that the magnet 230 be installed in the same direction as the rotation of the chamber (see Figure 2). When the chamber is separated from the magnet 230, it is preferable to minimize the influence of magnetism so that the magnetic nanoparticles float within the chamber. In this case, if the chambers are installed at equal intervals, and the rotation angle of the chamber is set to 1 / 2 the installation angle of the magnet 230, then each chamber will be arranged so that it is at equal intervals from each magnet 230. In this case, since the chamber is located between two magnets 230, the influence of magnetic force on the magnetic nanoparticles inside the chamber is minimized, and they may float within the chamber (see Figure 3).

[0102] If the magnet 230 is installed on the outside or inside of the direction in which the chamber rotates, then despite such rotation, the magnetic particles may be collected on the inside or outside of the chamber due to the influence of the magnet 230, as the magnet 230 is located on the inside or outside.

[0103] A rotating means 240 capable of rotating the upper plate may be installed on the outer surface of the upper plate. As described above, in the present invention, the distance between the chamber and the magnet 230 can be adjusted according to the rotation of the upper plate. For such rotation, it is preferable that a rotating means 240 is installed on the outer surface of the upper plate, and the rotating means 240 can rotate the upper plate in the forward or reverse direction by a force transmitted from a rotating power unit 300, which will be described later.

[0104] In this case, the rotating means 240 can be installed without limitation as long as it can rotate the upper plate, but preferably it may be a gear shape installed on the outer circumferential surface of the upper plate. That is, the upper plate of the present invention may have a cylindrical gear shape with gear teeth formed on its outer circumferential surface overall.

[0105] Furthermore, while the gear teeth are preferably made as spur gears, they may be made as bevel gears or screw gears depending on the installation position of the rotating power unit 300 described later, and they may also be made as helical gears to reduce noise.

[0106] The present invention also relates to a rapid sterility testing platform including a biochip for the rapid sterility testing platform.

[0107] The rapid sterility testing platform may include the biochip, a rotational power unit 300 for rotating the upper plate of the biochip, an illumination unit 400 installed above or below the biochip, and an image acquisition unit 500 for capturing images of the inside of the chamber of the biochip (see Figure 4).

[0108] Since the aforementioned biochip is the same as the one described above, its explanation will be omitted.

[0109] The rotational power unit 300 is the part that supplies power for the rotation of the upper plate and may be a gear connected to an electric motor. In this case, the gear may be directly connected to the rotating means 240 of the upper plate to supply power, or it may be connected via a chain and belt or the like. Furthermore, the gear installed in the rotational power unit 300 preferably has a shape corresponding to the rotating means 240 of the upper plate, and specifically may have the shape of a spur gear, helical gear, rack gear, bevel gear, screw gear, or worm gear.

[0110] An illumination unit 400 may be installed on the upper or lower part of the biochip. The illumination unit 400 is a part that supplies illumination for observing the inside of the chamber, and when observing changes in hue, white illumination can be used, and when observing fluorescence, ultraviolet or visible light can be supplied. Furthermore, it is preferable that the upper or lower surface 110 of the chamber be made of a transparent material for the illumination and observation described above.

[0111] The aforementioned illumination can be used without limitation as long as it supplies light of the desired hue, but preferably an LED light source can be used. In this case, the LED light source may be a light source that emits visible light or ultraviolet light, and a laser LED can also be used for fluorescence observation.

[0112] An image acquisition unit 500 may be installed to photograph the inside of the chamber of the biochip. The image acquisition unit 500 is used to observe changes in hue or fluorescence inside the chamber, and an image acquisition device using a CCD can be used. In the present invention, during sterility testing, the hue or fluorescence can be changed by an oxidation-reduction indicator as described above, so the presence or absence of microorganisms can be easily confirmed by acquiring images at regular time intervals using the image acquisition unit 500 as described above. Furthermore, as described above, in order to improve accuracy during image acquisition, the upper plate is rotated to bring the chamber and the magnet 230 closer together.

[0113] To describe the image acquisition unit 500 in detail, for continuous fluorescence imaging over time, the present invention can utilize a miniaturized imaging system measuring 224 mm × 120 mm × 140 mm. The imaging system includes an xyz motor-driven stage with adjustable imaging area and focus relative to the chip sample, a 4X objective lens, and a CCD camera, and may further include the illumination 400. Furthermore, it is preferable that the imaging system of the present invention be manufactured in a simplified form specifically for the purpose of rapid sterility verification. This is significantly cheaper than conventional commercially available equipment such as plate readers, can be widely adopted in industry in a simplified form, can be easily incorporated into incubators and the sterility testing platform of the present invention, and is advantageous for temperature maintenance and space optimization.

[0114] Preferred embodiments of the present invention will be described below with reference to the accompanying drawings so that they can be easily implemented by a person with ordinary skill in the art. Furthermore, in describing the present invention, if it is determined that a specific description of a related known function or configuration would unnecessarily obscure the gist of the invention, such detailed description will be omitted. Note that certain features shown in the drawings have been enlarged, reduced, or simplified for the sake of clarity, and the drawings and their components are not necessarily shown in appropriate proportions. However, a person skilled in the art will be able to easily understand these details.

[0115] Example 1

[0116] The isolation and concentration of microorganisms using nanoparticles is carried out through the process shown in Figure 3. After adding nanoparticles coated with a substance that binds to microorganisms, in proportion to the sample volume of the biopharmaceutical to be tested, the mixture is stirred at 35°C for approximately 15 minutes to induce binding between the nanoparticles and the microorganisms.

[0117] In this study, the nanoparticles were coated with apolipoprotein H, a type of protein. The nanoparticles were then attached to a magnetic bar for 5 minutes to isolate them against the wall, after which the supernatant was removed. Finally, a small amount of culture medium was added to the sample containing only nanoparticles bound to microorganisms, and the sample was resuspended. Specifically, tryptic soy broth was used as the culture medium for detecting aerobic microorganisms, and fluid thioglycolate medium was used for detecting anaerobic microorganisms.

[0118] The resuspended sample was injected into the sample chamber of the biochip, and 10% by weight of the redox indicator, Rezazlin, was added. The control chamber was filled with magnetic nanoparticles, culture medium, and Rezazlin at the same concentrations as the resuspended sample.

[0119] Subsequently, the biochip was inserted into the imaging system shown in Figure 4, and fluorescence imaging was performed at regular intervals. Using analytical software, the fluorescence signals extracted from the images taken from both the sample and a control group free of microorganisms were measured and their intensities were compared. If the fluorescence signal measured from the sample was significantly higher than the fluorescence signal measured from the control group, it was determined to be contaminated.

[0120] Experimental results

[0121] Figure 7 shows the experimental results confirming the performance of the biochip included in the present invention. Comparing the number of microorganisms after 12 hours with a control group (ctrl.) cultured in growth medium without nanoparticles, the number of microorganisms was approximately five times lower in the environment where magnetism was applied and nanoparticles aggregated (w / mag.). This indicates that the growth of microorganisms was inhibited due to the aggregated environment. On the other hand, when simulating an environment without magnetism (w / o mag.) using the biochip, the same number of microorganisms as the control group was observed after the same incubation period, and this was statistically verified through five repeated experiments. In other words, from the above results, it was confirmed that, in the case of the present invention, the culture rate can be adjusted by simple operations such as rotating the upper plate.

[0122] Figure 8 shows the results of a comparison between the rapid sterility verification test method (NEST) proposed in the present invention and a conventional 14-day culture-based sterility verification test method for performance evaluation. Two test methods were performed using a biopharmaceutical drug with a concentration of 1 CFU / mL for six representative microorganisms recommended for sterility verification performance evaluation. In 60 tests each, the rapid sterility verification test method of the present invention was confirmed to provide a measurement time that was approximately 18 times shorter on average compared to the conventional test method, along with even better positive result accuracy. Positive results were detected in 57 out of 60 cases with the rapid sterility verification test method (95% accuracy), while positive results were detected in 50 cases with the conventional sterility verification test method (83.3% accuracy).

[0123] Figure 9 shows the results of rapid sterility verification tests for various types of biopharmaceuticals. The rapid sterility verification test, which demonstrates superior performance compared to conventional methods, can be universally applied regardless of the substances contained in the biopharmaceutical, by specifically isolating only microorganisms. To verify this, a single type of microorganism was administered to various biopharmaceuticals, including clinical-grade CAR-T therapies, cell therapies, and RNA vaccines, and the rapid sterility verification test was applied. The results showed that all samples exhibited similar signal changes over time, demonstrating the ability to specifically detect only the signal for the microorganism, and confirming the expandability of the invented rapid sterility verification method to applicable industrial fields.

[0124] Furthermore, the rapid sterility verification test of the present invention allows for easy visual confirmation of the presence or absence of microorganisms, in addition to the fluorescence-based method described above. As shown in Figure 10, this method is not applicable to anaerobic microorganisms and takes relatively longer than fluorescence imaging analysis for clear comparisons, but it can be a means of easily enabling sterility verification tests even in environments without imaging equipment. For bacteria with a fast division rate, the presence or absence of biopharmaceutical contamination can be confirmed within 16 hours, and for fungal strains with a relatively slow division rate, within 24 hours.

[0125] As described above, while specific parts of the present invention have been described in detail, it is clear to those with ordinary skill in the art that these specific descriptions are merely preferred modes of implementation and do not limit the scope of the present invention. Therefore, the substantial scope of the present invention shall be defined by the appended claims and their equivalents.

Claims

1. A rapid sterility testing method using a biochip, comprising one or more chambers into which a sample and a control group are loaded, and one or more magnets installed on one side of each chamber, A step of mixing magnetic particles with the substance to be tested and bonding the analyte to the magnetic particles, The steps include separating magnetic particles from the substance to be tested to produce a sample, The steps include: injecting the separated sample and indicator substance into the biochip chamber and culturing them to determine the presence of the analyte; Includes, The step of determining the presence of the analyte is a rapid sterility test method in which, after culturing, the fluorescence or hue of the sample and the control group image is compared to determine the presence or absence of the analyte within the sample.

2. The rapid sterility test method according to claim 1, characterized in that the substance to be tested is a biopharmaceutical, cell therapy agent, clinical sample, chemical drug, cosmetic, food, or environmental sample.

3. The rapid sterility test method according to claim 1, characterized in that the analyte is bacteria, mycoplasma, or fungi.

4. The rapid sterility test method according to claim 1, characterized in that the step of producing the sample includes a step of concentrating the analyte bound to the magnetic particles on a growth medium.

5. The rapid sterility test method according to claim 4, wherein the growth medium comprises tryptic soy broth, Muller-Hinton broth, fluid thioglycolate medium, Sabouraud broth, and Luria-Bertani broth.

6. The step of determining the presence of the object to be analyzed is: The steps include separating the chamber from the magnet, The steps include injecting the indicator substance, sample, and control group into the chamber and culturing for a certain period of time, After the cultivation is complete, the step is to bring the chamber closer to the magnet, After capturing an image of the inside of the chamber, the images are compared with those captured from the separated sample and the control group. A rapid sterility test method according to claim 1, including the method described in claim 1.

7. The analyte combines with the magnetic particles to form a composite, The rapid sterility test method according to claim 6, wherein the composite is moved to one side of the chamber by the magnet when the chamber is brought close to the magnet.

8. The rapid sterility test method according to claim 1, wherein the culture is performed for 4 to 48 hours.

9. The rapid sterility test method according to claim 1, wherein the image acquisition is performed at intervals of 10 to 120 minutes.

10. The rapid sterility test method according to claim 1, wherein the indicator substance changes fluorescence or hue depending on the analyte.

11. The rapid sterility test method according to claim 10, characterized in that the indicator substance is a redox indicator, a pH indicator, a dye-based reagent, an enzyme reaction indicator, or an antibody-based reagent.

12. A lower plate containing a chamber into which the control group and samples are loaded, The upper plate, which fits into the lower plate, contains a magnet, In biochips including, The aforementioned lower plate is The bottom and, The central shaft protruding from the bottom surface upwards, One or more chambers are installed at a certain distance from the central axis, Includes, The aforementioned upper plate is The central shaft insertion port into which the central shaft of the lower plate is inserted, The chamber insertion port into which the chamber is inserted, A magnet installed on one side of the chamber insertion opening, Biochips for rapid sterility testing platforms, including...

13. The biochip for a rapid sterile testing platform according to claim 12, wherein the chamber comprises one or more sample chambers and one or more control group chambers.

14. The biochip for a rapid sterile testing platform according to claim 13, wherein the chambers are installed at equal intervals at uniform distances from the central axis.

15. The biochip for a rapid sterile testing platform according to claim 12, further comprising magnetic particles and an indicator material inside the chamber.

16. The biochip for a rapid sterile testing platform according to claim 15, wherein the magnetic particles bind to the analyte to form a complex.

17. The biochip for a rapid sterility testing platform according to claim 16, wherein the magnetic particles include a functional group or substance on their surface that can bind to microorganisms.

18. The biochip for a rapid sterile testing platform according to claim 16, wherein the composite is positioned on one side of the chamber using the magnet when fluorescence or optical observation is performed using the indicator substance.

19. The biochip for a rapid sterile testing platform according to claim 12, wherein the upper plate is rotated at a constant angle about the central axis.

20. The biochip for a rapid sterile testing platform according to claim 19, wherein the control group chamber insertion portion has an arc shape centered on the central axis.

21. The biochip for a rapid sterility testing platform according to claim 19, wherein the distance of the magnet to the chamber changes in accordance with the rotation of the upper plate.

22. The biochip for a rapid sterile testing platform according to claim 19, wherein a rotating means for rotating the upper plate is provided on the outer surface of the upper plate.

23. A rapid sterility testing platform comprising a biochip for a rapid sterility testing platform according to any one of claims 12 to 22.

24. The aforementioned rapid sterility testing platform is The aforementioned biochip, A rotational power unit for rotating the upper plate of the biochip, A lighting unit installed on the upper or lower part of the biochip, An image acquisition unit for capturing images of the inside of the chamber of the biochip, A rapid sterility testing platform according to claim 23, including the above.

25. The rapid sterility testing platform according to claim 24, wherein the image acquisition unit acquires a fluorescent image or optical image of the inside of the chamber.

26. The rapid sterility testing platform according to claim 24, wherein the rapid sterility testing platform determines the presence or absence of microorganisms in the sample by comparing the fluorescence or hue of the initial image obtained with the current image.

Images