A clinical laboratory automation system using a single calibrator.

Abstract

External calibration curves rely on external calibrators containing known concentrations of target analytes that may degrade over time, leading to inaccurate results. Generating a new calibration curve often requires preparing several calibrators to obtain the calibration points required to generate the calibration curve. Preparing the calibrators required for a multi-point calibration curve requires operator preparation time and may result in handling errors. The claimed and described technology provides a clinical laboratory automation system that includes a fluid handling system, an analyzer component, and a mass spectrometer. The clinical laboratory automation system can provide automated calibration using one calibrator to prepare one or more calibrator dilutions used to generate a calibration curve for the quantitative measurement of a target analyte in a sample.

Description

[Technical Field]

[0001] (Related applications) This patent application claims priority benefit of U.S. Provisional Patent Application No. 63 / 131,927, filed on December 30, 2020, the contents of which are incorporated as a whole within this disclosure by reference.

[0002] (Field) Various aspects of this disclosure relate to automated calibration using one calibrator to prepare one or more calibrator dilutions used to generate a calibration curve for quantitative measurement of a target specimen in a sample. Other aspects of the invention provide automated evaluation of pipette dispensing volumes and adjustment of pipette actuators for delivering accurate dispensing volumes. [Background technology]

[0003] Mass spectrometry (MS) is an analytical technique used to determine the elemental composition of a sample, quantify the mass of particles and molecules, and elucidate the chemical structure of molecules. Various types of MS with high specificity, such as liquid chromatography (LC-MS), gas chromatography (GC-MS), and matrix-assisted laser desorption / ionization / time-of-flight (MALDI-TOF MS), are increasingly being used in clinical diagnostics. These MS techniques overcome many of the limitations of immunological assays (e.g., nonspecific binding and cross-reactivity of samples) and offer numerous advantages.

[0004] Quantification by MS can be performed using an external calibration curve. An external calibration curve relies on an external calibrator containing a target sample at a known concentration. These calibrators degrade over time, potentially leading to inaccurate results. Generating a new calibration curve often requires preparing several calibrators and obtaining the calibration points necessary to generate the curve. Preparing the calibrators required for a multi-point calibration curve requires operator preparation time and can lead to handling errors. For example, some assays require at least a five-point external calibration curve.

[0005] Embodiments of the present invention address these calibration challenges and other challenges individually and collectively.

[0006] Further limitations and shortcomings of conventional and traditional approaches will become apparent to those skilled in the art through a comparison of such systems with some aspects of this disclosure, as described in the remainder of this application with reference to the drawings. [Overview of the Initiative] [Means for solving the problem]

[0007] One aspect provides automated calibration using one calibrator to prepare one or more calibrator dilutions used to generate a calibration curve for quantitative measurement of a target specimen in a sample. Several aspects of the present invention provide automated evaluation of pipette dispensing volume and adjustment of pipette actuator for delivering accurate dispensing volumes.

[0008] Other aspects include a clinical laboratory automation system comprising (i) a fluid handling system comprising a container handler, at least one fluid container, and a pipette arrangement; (ii) an analytical component; and (iii) a mass spectrometer. The fluid handling system is configured to dispense at least one fluid from the pipette arrangement into at least one fluid container. The mass spectrometer is configured to at least evaluate the properties of at least one fluid, thereby producing corresponding sets of values. The corresponding sets of values ​​can be used to calibrate the analytical component. The clinical laboratory automation system may include a control system configured to control the fluid handling system, the analytical component, and the mass spectrometer.

[0009] Some aspects may include an integrated clinical laboratory automation system that includes an analytical component integrated with a mass spectrometer and a fluid handling system integrated with the analytical component and / or the mass spectrometer. In some embodiments, the analytical component includes an immunological assay analyzer, a clinical chemistry analyzer, a protein chemistry analyzer, a hematology analyzer, or a urinalysis analyzer.

[0010] Another aspect concerns a method for calibrating an immunoassay analyzer or clinical chemistry analyzer, the method being carried out by a clinical laboratory automation system comprising (i) a fluid handling system comprising a container handler, at least one fluid container, and a pipette arrangement, (ii) an analyzer component, and (iii) a mass spectrometer. In one embodiment, the method includes dispensing a first required volume of diluent fluid from a first pipette in the pipette arrangement of the fluid handling system into a fluid container, dispensing a second required volume of calibrator from a second pipette in the pipette arrangement of the fluid handling system into the same fluid container to produce a dilution series containing at least one dilution of the calibrator, performing an evaluation of the concentration of at least one dilution of the calibrator from the dilution series using a mass spectrometer, generating corresponding sets of values, thereby generating an RLU dose calibration curve, and calibrating the immunoassay analyzer or clinical chemistry analyzer at least in part using the RLU dose calibration curve. In an alternative method, the immunological assay analyzer or clinical chemistry analyzer includes an RLU dose master calibration curve, and the corresponding values ​​are used to adjust the RLU dose master calibration curve, thereby calibrating the immunological assay analyzer or clinical chemistry analyzer.

[0011] Further aspects concern a method for calibrating an immunoassay analyzer or clinical chemistry analyzer, the method being carried out by a clinical laboratory automation system comprising (i) a container handler, a fluid handling system comprising at least one fluid container and a pipette arrangement, (ii) a sample pipette station, (iii) an analyzer component, and (iv) a mass spectrometer. The method includes dispensing a first required volume of diluent fluid from a first pipette in the pipette arrangement of the fluid handling system into the fluid container, dispensing a required volume of calibrator from the sample pipette station into the same fluid container to produce a dilution series containing at least one dilution of the calibrator, performing an evaluation of the concentration of at least one dilution of the calibrator from the dilution series using a mass spectrometer, generating corresponding sets of values, thereby generating an RLU dose calibration curve, and calibrating the immunoassay analyzer or clinical chemistry analyzer at least in part using the RLU dose calibration curve.

[0012] Another aspect concerns a method for adjusting pipette dispensing volume, the method being carried out by a clinical laboratory automation system comprising (i) a container handler, a pipette arrangement comprising at least one fluid container, at least a first pipette, and a second pipette, and a fluid handling system comprising at least one pump driven by an actuator and associated with the first and / or second pipette, (ii) an analyzer component, and (iii) a mass spectrometer. The method includes dispensing a first required volume of a first diagnostic reagent containing a sample from the first pipette into the fluid container, dispensing a second required volume of a second diagnostic reagent containing an antibody from the second pipette into the same fluid container, quantifying the mixture of diagnostic reagents by mass spectrometer, generating corresponding sets of values, evaluating with respect to pipette dispensing inaccuracy using at least corresponding sets of values, and, if dispensing inaccuracy is determined, adjusting the actuator as necessary to dispense the correct pipette dispensing volume.

[0013] Another aspect concerns a method for providing variable dilution of a fluid, the method being carried out by a laboratory automation system comprising a fluid handling system comprising a container handler and a pipette arrangement comprising at least a first fluid container, at least a first pipette and a second pipette, the fluid handling system being configured to produce a set of dilution series of a calibrator. The method comprises providing a calibrator, providing a diluent, dispensing a first required volume of the diluent from the first pipette into the first fluid container, and dispensing a first required volume of the calibrator from the second pipette into the same fluid container.

[0014] A further aspect concerns a method for providing variable dilution of a fluid, the method being carried out by a clinical laboratory automation system comprising a fluid handling system comprising a sample pipette station, a container handler, at least a first fluid container, and a pipette arrangement having at least a first pipette, the fluid handling system being configured to produce a set of dilution series of a calibrator. The method comprises providing a calibrator, providing a diluent, dispensing a first required volume of the diluent from a first pipette into a first fluid container, and dispensing a required volume of the calibrator from a sample pipette station into the same fluid container.

[0015] These and other embodiments of the present invention are described in further detail below with reference to the drawings. The present invention provides, for example, the following: (Item 1) A clinical laboratory automation system (100), wherein the clinical laboratory automation system (100) is A fluid handling system (104) comprising a container handler (101), at least one fluid container (103), and a pipette arrangement (421), wherein the fluid handling system (104) is configured to dispense at least one fluid into the at least one fluid container, Analysis component (102), A mass spectrometer (106) configured to evaluate the properties of at least one of the fluids and thereby produce a corresponding set of values Equipped with, The analytical component (102) is calibrated at least partially using the corresponding set of values ​​in the automated clinical laboratory system (100). (Item 2) The automated clinical laboratory system (100) according to item 1, further configured to evaluate the dispensing inaccuracy of the pipette arrangement (421) using at least a corresponding set of the aforementioned values. (Item 3) The analytical component (102) is a clinical laboratory automation system (100) as described in item 1 or 2, including an immunoassay analyzer. (Item 4) The analytical component (102) is a clinical laboratory automation system (100) as described in item 1 or 2, which includes a clinical chemistry analyzer, a protein chemistry analyzer, a hematology analyzer, or a urinalysis analyzer. (Item 5) The aforementioned automated clinical laboratory system is an automated clinical laboratory system (100) according to any one of items 1-4, further comprising a sample pipette station (402a). (Item 6) The pipette arrangement (421) comprises a pump (414), the pump (414) being driven by an actuator (415), the clinical laboratory automation system (100) according to any one of items 1-5. (Item 7) The actuator is a motor, as described in item 6 of the clinical laboratory automation system (100). (Item 8) The motor is a stepper motor, as described in item 7 of the clinical laboratory automation system (100). (Item 9) The at least one fluid is selected from the group consisting of calibrators, diagnostic reagents, diluents, or mixtures thereof, in the automated clinical laboratory system (100) according to any one of items 1-8. (Item 10) The calibrator is a clinical laboratory automation system (100) as described in item 9, comprising at least one type of sample or antibody. (Item 11) The calibrator comprises multiple different types of specimens and / or antibodies, as described in item 9 of the automated clinical laboratory system. (Item 12) The calibrator or diagnostic reagent comprises a sample, the sample being independently selected from the group consisting of thyroid-stimulating hormone (TSH), prostate-specific antigen (PSA), troponin, vitamin D, and free thyroxine (T4), according to item 9 (100). (Item 13) The diagnostic reagent comprises an antibody or antigen, as described in item 9, for the automated clinical laboratory system (100). (Item 14) The diluent is TRIS buffer or bovine serous albumin (BSA) buffer, as described in item 9 (100). (Item 15) The corresponding set of the aforementioned values ​​is a clinical laboratory automation system (100) described in any of items 1-14, which includes a single value. (Item 16) The pipette arrangement further comprises at least a first pipette (404), the first pipette (404) configured to dispense at least one required volume of a fluid, as described in any one of items 5-15, the clinical laboratory automation system (100). (Item 17) The fluid handling system (104) is configured to dispense at least two fluids, preferably a first fluid and a second fluid, as described in item 16, for the automated clinical laboratory system (100). (Item 18) The first pipette (404) is configured to dispense either a first required volume of the first fluid or a second required volume of the second fluid into the same fluid container, as in the clinical laboratory automation system (100) described in item 17. (Item 19) The pipette arrangement further comprises at least a second pipette (405), wherein the first pipette (404) is configured to dispense a first required volume of a first fluid into a fluid container, and the second pipette (405) is configured to dispense a second required volume of a second fluid into the same fluid container, as described in item 16 or 17, for the clinical laboratory automation system (100). (Item 20) The automated laboratory system (100) according to item 16, wherein the first pipette (404) is configured to dispense a first required volume of a first fluid into a fluid container, and the sample pipette station (402a) is configured to dispense a second required volume of a second fluid into the same fluid container. (Item 21) A clinical laboratory automation system (100) according to any one of items 17-20, wherein the first fluid is the diluent and the second fluid is the calibrator. (Item 22) The fluid handling system (104) is configured to produce a dilution series of the calibrator, the dilution series comprising at least one dilution of the calibrator, according to item 21, the clinical laboratory automation system (100). (Item 23) The clinical laboratory automation system (100) according to item 22, wherein the characteristic to be evaluated is the concentration of the at least one dilution of the calibrator from the set of dilution series of the calibrator. (Item 24) A clinical laboratory automation system (100) according to item 23, wherein an RLU dose conversion curve is generated from a corresponding set of the aforementioned values, and the analytical component is calibrated at least partially using the RLU dose conversion curve. (Item 25) The automated clinical laboratory system (100) described in item 23 includes an RLU dose master calibration curve, wherein the corresponding set of values ​​is used to generate an adjusted calibration curve. (Item 26) A clinical laboratory automation system (100) as described in any one of items 17-19, wherein the first fluid is a first diagnostic reagent and the second fluid is a second diagnostic reagent. (Item 27) The first diagnostic reagent comprises a specimen, and the second diagnostic reagent comprises an antibody or antigen, according to item 26 (100). (Item 28) The characteristic to be evaluated is the quantification of a mixture of the first diagnostic reagent and the second diagnostic reagent, as described in item 27 (100) of the automated clinical laboratory system. (Item 29) The quantification of the mixture of diagnostic reagents comprises quantifying the mixture of the first and second diagnostic reagents by molecular weight shift, according to the automated clinical laboratory system (100) of item 28. (Item 30) If the aforementioned dispensing inaccuracy is determined, the motor step of the actuator (107) is adjusted as necessary to dispense the respective volumes, as described in item 29 of the automated laboratory system (100). (Item 31) If the aforementioned dispensing inaccuracy is determined, the pipette configuration is calibrated at least partially using the corresponding set of values, according to the clinical laboratory automation system (100) described in item 29. (Item 32) A clinical laboratory automation system (100) according to any one of items 1-31, wherein at least a portion of the fluid handling system (104) is integrated with the analytical component (102). (Item 33) A clinical laboratory automation system (100) according to any one of items 1-32, wherein at least a portion of the fluid handling system (104) is integrated with the mass spectrometer (106). (Item 34) The clinical laboratory automation system (100) according to any one of items 1-33 further comprises a control system (108) configured to control the fluid handling system (104), the analytical component (102), and / or the mass spectrometer (106). (Item 35) A method for calibrating an immunoassay analyzer or a clinical chemistry analyzer, wherein the method is: The step includes providing a clinical laboratory automation system (100), The aforementioned automated clinical laboratory system (100) (a) A fluid handling system (104) comprising a container handler (101), at least one fluid container (103), and a pipette arrangement (421), wherein the pipette arrangement (421) comprises at least a first pipette (404) and at least a second pipette (405), The first pipette (404) is configured to dispense a first required volume of the first fluid into the fluid container (103), and the second pipette (405) is configured to dispense a second required volume of the second fluid into the same fluid container (103). The first fluid is a diluent, and the second fluid is a calibrator. The fluid handling system (104) is configured to produce at least a dilution series of the second fluid, and the dilution series comprises at least one diluent of at least the second fluid, (b) an immunoassay analyzer or clinical chemistry analyzer, (c) A mass spectrometer (106) configured to evaluate the concentration of at least one dilution from the set of dilution series of the calibrator and generate a corresponding set of values. Equipped with, A method wherein an RLU dose conversion curve is generated from a corresponding set of the aforementioned values, and the immunoassay analyzer or the clinical chemistry analyzer is calibrated, at least in part, using the RLU dose conversion curve. (Item 36) A method for adjusting the pipette dispensing volume, wherein the method is The step includes providing a clinical laboratory automation system (100), The aforementioned automated clinical laboratory system (100) (a) A fluid handling system (104) comprising a container handler (101), at least one fluid container (103), and a pipette arrangement (421), wherein the pipette arrangement (421) comprises at least a first pipette (404), at least a second pipette (405), and a pump (414), the pump (414) being driven by an actuator (415), The first pipette (404) is configured to dispense a first required volume of the first fluid into the fluid container, and the second pipette (405) is configured to dispense a second required volume of the second fluid into the same fluid container (103). The fluid handling system (104) comprises the following: the first fluid is a first diagnostic reagent, the second fluid is a second diagnostic reagent, the first diagnostic reagent contains a sample, and the second diagnostic reagent contains an antibody or antigen. (b) an immunoassay analyzer or clinical chemistry analyzer, (c) A mass spectrometer (106) configured to quantify a mixture of the first and second diagnostic reagents, thereby producing a corresponding set of values. Equipped with, The method further comprises a clinical laboratory automation system (100) configured to evaluate the dispensing inaccuracy of the pipette arrangement (421) using at least a corresponding set of the aforementioned values, and if dispensing inaccuracy is determined, the actuator (415) is adjusted as necessary to dispense the respective volumes. (Item 37) A method for providing variable dilution of a fluid, wherein the method is To provide a fluid handling system (104), the fluid handling system (104) comprising a container handler (101), at least a first fluid container (103), and a pipette arrangement (421), wherein the pipette arrangement comprises at least a first pipette (404) and a second pipette (405), and the fluid handling system (104) is configured to produce a set of dilution series of calibrators. To provide a calibrator, To provide a diluent, Diluting the aforementioned calibrator in a diluent and Includes, Diluting the aforementioned calibrator in a diluent is, Using the pipette arrangement (421), the required first volume of the diluent is dispensed into the first fluid container (103), Using the pipette arrangement (421), the required first volume of the calibration agent is dispensed into the first fluid container (103). A method by which. (Item 38) The fluid handling system (104) includes at least a second fluid container (103a), and the method is Using the aforementioned pipette arrangement (421), the required second volume of the diluent is dispensed into the second fluid container. Using the pipette arrangement (421), the required second volume of the calibration agent is dispensed into the second fluid container (103a). The method described in item 37, further including the method described in item 37. (Item 39) The fluid handling system (104) includes at least a third fluid container (103b), and the method is Using the pipette arrangement (421), the required third volume of the diluent is dispensed into the third fluid container (103b), Using the pipette arrangement (421), the required third volume of the calibration agent is dispensed into the third fluid container (103b). The method described in item 38, further including the method described in item 38. (Item 40) The fluid handling system (104) includes at least a fourth fluid container (103c), and the method is Using the pipette arrangement (421), the required fourth volume of the diluent is dispensed into the fourth fluid container (103c), Using the pipette arrangement (421), the required fourth volume of the calibration agent is dispensed into the fourth fluid container (103c). The method described in item 39, further including the method described in item 39. (Item 41) A method for calibrating an immunoassay analyzer or a clinical chemistry analyzer, wherein the method is: A step of providing a clinical laboratory automation system (100), wherein the clinical laboratory automation system (100) is (a) A fluid handling system (104) including a container handler (101), at least one fluid container (103), and a pipette arrangement (421) having at least a first pipette (404), (b) Sample pipette station (402a), (c) an immunoassay analyzer or clinical chemistry analyzer, (d) Mass spectrometer (106) and It has steps, The steps include: dispensing a first required volume of diluent fluid from the first pipette (404) in the pipette arrangement (421) into the fluid container (103); The steps include: dispensing a required volume of the calibrator from the sample pipette station (402a) into the same fluid container to produce a dilution series containing at least one dilution of the calibrator; The mass spectrometer (106) is used to evaluate the concentration of at least one dilution of the calibrator from the dilution series, generate corresponding sets of values, and thereby generate an RLU dose calibration curve. The steps include calibrating the immunoassay analyzer or clinical chemistry analyzer using, at least partially, the RLU dose calibration curve, and Methods that include... (Item 42) A method for providing variable dilution of a fluid, wherein the method is To provide a sample pipette station (402a) and a fluid handling system (104), wherein the fluid handling system (104) includes a container handler (101), at least a first fluid container (103), and a pipette arrangement (421), the pipette arrangement comprising at least a first pipette (404), and the fluid handling system (104) is configured to produce a set of dilution series of calibrators. To provide a calibrator, To provide a diluent, Diluting the aforementioned calibrator in a diluent and Including, before Diluting the calibration agent in a diluent is Using the first pipette (404) in the pipette arrangement (421), the required first volume of the diluent is dispensed into the first fluid container (103), Using the sample pipette station (402a), the required volume of the calibration agent is dispensed into the first fluid container (103). A method by which. [Brief explanation of the drawing]

[0016] Embodiments of the present disclosure will be described hereby only by reference to the accompanying figures.

[0017] [Figure 1] Figure 1 shows a block diagram of an automated clinical laboratory system according to one embodiment of the present invention.

[0018] [Figure 2A] Figure 2A shows a schematic diagram of an analyzer component in an automated clinical laboratory system according to one embodiment of the present invention.

[0019] [Figure 2B] Figure 2B shows a schematic diagram of an alternative embodiment of an analyzer component in a clinical laboratory automation system according to another embodiment of the present invention.

[0020] [Figure 3] Figure 3 shows an illustrative flowchart illustrating the operating procedure for running a fluid handling system to prepare a calibration dilution series.

[0021] [Figure 4] Figure 4 shows an illustrative flowchart illustrating the operating procedure for running a fluid handling system to prepare a reagent mixture for evaluating pipette dispensing volume.

[0022] [Figure 5] Figure 5 shows a block diagram of the mass spectrometer.

[0023] [Figure 6] Figure 6 shows a part of a mass spectrometer that uses the electrospray method.

[0024] [Figure 7] Figure 7 shows the structure of an ion detector used in a mass spectrometer.

[0025] [Figure 8] Figure 8 shows a flowchart illustrating the calibration curve formation process according to one embodiment of the present invention.

[0026] [Figure 9] Figure 9 shows a flowchart illustrating a process for evaluating and adjusting pipette dispensing volume according to one embodiment of the present invention.

[0027] [Figure 10A] Figure 10A shows the extent to which the calibration signal of a single thyroid-stimulating hormone calibrator dilution curve can degrade over time.

[0028] [Figure 10B] Figure 10B shows a single thyroid-stimulating hormone calibrated dilution curve prepared according to one embodiment of the present invention.

[0029] [Figure 10C] Figure 10C shows the adjustment of the master calibration curve according to an alternative embodiment of the present invention.

[0030] [Figure 11] Figure 11 shows the analysis of diagnostic reagents using a mass spectrometer. [Modes for carrying out the invention]

[0031] Various embodiments will be described in detail with reference to drawings in which similar reference numerals represent similar parts and assemblies throughout several figures. It should be understood that this disclosure is not limited to and is therefore subject to variation of the specific methodologies, protocols, and reagents described herein. It should also be understood that the terminology used herein is for the purpose of describing specific embodiments only and is not intended to limit the scope of this disclosure or the appended claims.

[0032] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly indicates otherwise.

[0033] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which this disclosure pertains.

[0034] Some embodiments may be used to calibrate analyzers used to detect the presence, absence, or concentration of a sample in a biological or chemical sample. Biological samples, such as biological fluids, may include, but are not limited to, blood, plasma, serous fluid, or other bodily fluids or excretions (including, but not limited to, saliva, urine, cerebrospinal fluid, tears, sweat, gastrointestinal fluid, amniotic fluid, mucosal fluid, pleural fluid, sebum, breath, etc.). Chemical samples may include any suitable type of sample containing a chemical substance (including water samples).

[0035] Prior to discussing some embodiments, some terms may be explained in more detail.

[0036] The term “analyzer” or “analytical component” may include any suitable instrument capable of analyzing a sample such as a component, fluid, or biological sample. Examples of analyzers or analytical components include mass spectrometers, immunological assay analyzers, hematology analyzers, microbiology analyzers, and / or molecular biology analyzers.

[0037] In some embodiments, the analyzer may be an immunological assay analyzer (typically detecting labels (chemiluminescence, electrochemiluminescence fluorescence, radioactivity, isotopes, DNA, etc.)) or a label-free system. Other types of analyzers may include hematological analyzers, microbiological analyzers, chemical analyzers, urine analyzers, biochemical analyzers, and / or molecular biology analyzers. When analyzing a biological sample, one or more of these types of analyzers in any suitable combination may be used to analyze the biological sample.

[0038] Hematological analyzers can be used to perform whole blood count, erythrocyte sedimentation rate (ESR), and / or coagulation tests. Automated cell counters sample blood and quantify, classify, and describe the cell population using both electrical and optical techniques.

[0039] Microbiological analyzers can function as diagnostic tools for determining the identification of biological organisms. In some embodiments, microbiological analyzers can identify infectious microorganisms. Depending on the type of test performed, such analyzers can use multiple small sample testing microwells or multiwell panels containing different substrates within a centrifugal rotor to analyze biochemicals.

[0040] A molecular biology analyzer can be a device capable of analyzing a biological sample at its molecular level. Examples of molecular biology analyzers may include nucleic acid analyzers such as DNA analyzers.

[0041] Chemical analyzers can perform assays on clinical samples such as serum, plasma, urine, and cerebrospinal fluid to detect the presence of disease- or drug-related samples. Chemical analyzers may use photometric methods. In photometric methods, the sample is mixed with appropriate reagents to produce a reaction that yields a color. The concentration of the sample determines the intensity of the color produced. A photometer emits light of the appropriate wavelength in the sample and measures the amount of absorbed light, which is directly related to the concentration of the sample in the sample. Another analytical method used within chemical analyzers is Na + , K + Ca + F - Cl - , and Li + This involves the use of ion-selective electrodes (ISEs) to measure ions such as [specific ions]. An ISE is a sensor that determines the concentration of ions in a solution by measuring the current flow through an ion-selective membrane.

[0042] The term "analyte" can include substances whose presence, absence, or concentration is to be determined according to embodiments of the present invention. Exemplary analytes include, but are not limited to, organic molecules, hormones (such as thyroid hormones, estradiol, testosterone, progesterone, estrogen, etc.), metabolites (such as glucose or ethanol, etc.), proteins, lipids, carbohydrates, and glycans, steroids (such as vitamin D, etc.), peptides (such as procalcitonin, etc.), nucleic acid segments, biomarkers (such as pharmaceuticals like antibiotics, benzodiazepines, etc.), drugs (such as immunosuppressants, narcotics, opioids, etc.), molecules having an adjustment effect in enzymatic processes such as promoters, activators, inhibitors, or cofactors, microorganisms (including viruses such as EBV, HPV, HIV, HCV, HBV, influenza, norovirus, rotavirus, adenovirus, etc.), bacteria (such as Helicobacter pylori, Streptococcus, MRSA, Clostridium difficile, Legionella, etc.), fungi, parasites (such as Plasmodium malariae, etc.), cells, cell components (such as cell membranes, etc.), spores, nucleic acids (such as DNA and RNA, etc.), etc. Embodiments of the present invention can also enable simultaneous analysis of multiple analytes in the same class or different classes (e.g., simultaneous analysis of metabolites and proteins). In embodiments of the present invention, analysis of a specific analyte such as a biomarker can indicate that a specific medical condition (e.g., a disease) is associated with a sample containing the analyte.

[0043] The term "immunological assay" refers to laboratory methods used to determine the amount of an analyte in a sample. It can be based on the interaction between an antibody and an antigen, and depending on the degree of selectivity for the analyte (either antigen or antibody), an immunological assay can be used to quantitatively determine very low concentrations of the analyte in a test sample. An "immunoassay analyzer" or "immunological assay analyzer" can include an instrument in which the immunological assay based thereon is automated. Dxl TM System (Beckman Coulter, CA), AD VIA TM and CENTAUR TM System (Siemens Healthcare, Germany), COB AS TMSystem (Roche Diagnostic, Germany), ARCHITECT TM System (Abbott, IL), VITROS TM The system (Ortho-clinical Diagnostic, NJ), and VIDAS TM Various immunoassay analyzers, including the Biomerieux system (France), are commercially available.

[0044] A "mass spectrometer" is an instrument capable of measuring mass and the relative concentrations of atoms and molecules. One example of a mass spectrometer utilizes the fundamental magnetic force on a moving charged particle. Essentially, the instrument ionizes the sample and then deflects the ions through a magnetic field based on the ion's mass-to-charge ratio. The mass spectrum can then be used to determine the elemental or isotopic characteristics of the sample, the mass of particles and molecules, and to elucidate the chemical structure of molecules such as peptides and other chemical compounds. Commercially available mass spectrometers can be categorized based on the method of mass selection, including time-of-flight, quadrupole MS, ion traps (including 3D quadrupole, cylindrical ion trap, linear quadrupole ion trap, and orbit trap), and Fourier transform ion cyclotron resonance (FTMS). Alternatively, they can also be categorized based on the ion source (laser desorption, matrix-assisted laser desorption, thermal ionization, plasma, spark source, etc.) or detector (electron multiplier tubes (Faraday cups and ion / photon detectors, etc.), induction detectors, etc.). In a preferred embodiment, the mass spectrometer may be a triple quadrupole mass spectrometer.

[0045] The term "calibration" refers to the process of determining the relationship between the instrument response (measured response) and the known sample concentration to ensure effective quantification of the sample.

[0046] The term "calibration curve" refers to the mathematical relationship between a measured response and a known sample concentration. Calibration curves are used to convert relative light units (RLU) measurements of a sample into specific quantitative sample concentrations.

[0047] In the specific examples provided below, a clinical laboratory automation system including a fluid handling system, an immunoassay analyzer, and a mass spectrometer is described in detail. However, embodiments of the present invention are not limited thereto. Another type of analyzer, such as a chemical analyzer, may be used instead of an immunoassay analyzer. Many of the functions and features found in an immunoassay analyzer may also be found in a chemical analyzer (e.g., reagent storage, preparative station, sample preparation station, etc.). Furthermore, additional components such as sample introduction devices may also be used in conjunction with the chemical analyzer and mass spectrometer within the clinical laboratory automation system.

[0048] Specific embodiments may include a fluid handling system, which may be separate from or integrated with the analytical components. The fluid handling system may also be separate from or at least partially integrated with the mass spectrometer. In some embodiments, the fluid handling system, analytical components, and mass spectrometer are individual components within a modular laboratory automation system. The modular laboratory automation system may have a workflow comprising a pre-analysis section, a post-analysis section, and at least one connection to the analytical components. The pre-analysis section may include a batch loading component, at least one centrifuge, and / or a sample quality detection component. The pre-analysis section may also include a fluid handling system. The post-analysis section may include a volume detection component and / or a storage and recovery component. Recovery may be automated or manual. Direct tracking sampling may be used, for example, to connect to an immunological assay analyzer or coagulation instrument. Rack construction units may be used, for example, to connect to a clinical chemistry analyzer or hematology analyzer. The mass spectrometer may be offline from the workflow or connected through other analytical connectors known in the art.

[0049] In some embodiments, the fluid handling system, analytical components, and mass spectrometer can be a fully integrated platform. In some embodiments, the fluid handling system is part of a sample preparation station. In some embodiments, the automated system includes a sample introduction station that can transfer samples to the mass spectrometer for analysis. The laboratory automated system also includes a control system that can control the fluid handling system, analytical components, and mass spectrometer.

[0050] Figure 1 shows a high-level block diagram of an automated clinical laboratory system according to one embodiment of the present invention. The automated clinical laboratory system 100 comprises an analyzer component (e.g., an immunoassay analyzer) 102, a mass spectrometer 106, and a fluid handling system 104. In some embodiments, the fluid handling system 104 may be integrated with a sample processing system. An automated sample processing system is described in detail in published PCT application WO2018 / 217778, published on November 29, 2018 (which is incorporated herein by reference as a whole). The fluid handling system 104 may be physically and / or operably coupled to the analyzer 102 and the mass spectrometer 106, and in some embodiments, the fluid handling system 104, the analyzer component 102, and the mass spectrometer 106 may form a single instrument. The fluid handling system 104 may play a role in preparing calibrator dilutions for quantitative calibration of the analyzer component and / or preparing reagent mixtures for evaluating pipette dispensing volumes. The calibrator dilution and reagent mixture can be transferred to the mass spectrometer 106 for analysis.

[0051] The analyzer component 102 may include an immunological assay analyzer, a clinical chemistry analyzer, a protein chemistry analyzer, a hematology analyzer, or a urinalysis analyzer. The analyzer component 102 may include several sample aliquot processing devices for forming processed sample aliquots for analysis. Such processing devices may process samples or sample aliquots in any preferred manner. Examples of sample aliquot processing devices include a reagent dosing station (e.g., a reagent pipetting station), a sample pipetting station, an incubator, a washing station (e.g., a magnetic washing station), a sample storage unit, etc. The analyzer component 102 may be an automated analyzer component 400.

[0052] A control system 108 may also be present within the clinical laboratory automation system 100. The control system 108 can control the analyzer component 102, the fluid handling system 104, and / or the mass spectrometer 106. The control system 108 may comprise a data processor 108A, a non-transient computer-readable medium 108B, and a data storage device 108C coupled to the data processor 108A. The non-transient computer-readable medium 108B may contain code executable by the data processor 108A to perform the functions described herein. The data processor 108C may store data for processing samples, sample data, or data for analyzing sample data.

[0053] The data processor 108A may include any suitable data computing device or combination of such devices. An exemplary data processor may comprise one or more microprocessors cooperating to perform a desired function. The data processor 108A may include a CPU having at least one fast data processor suitable for executing program components to perform user and / or system-generated requests. The CPU may be a microprocessor such as AMD Athlon, Duron, and / or Opteron, IBM and / or Motorola PowerPC, IBM and Sony Cell processors, Apple M1, Intel Celeron, Itanium, Pentium®, Xeon, and / or XScale processors.

[0054] The computer-readable medium 108B and the data storage device 108C may be any suitable device or a set of devices capable of storing electronic data. Examples of memory may comprise one or more memory chips, disk drives, etc. Such memory may operate using any suitable electrical, optical, and / or magnetic mode of operation.

[0055] The computer-readable medium 108B may contain code executable by the data processor 108A for performing any preferred method. For example, the computer-readable medium 108B may contain code executable by the processor 108A for automatically generating a calibration curve in the laboratory automation system using calibration dilution measurements from the mass spectrometer 106. In another embodiment, the computer-readable medium 108B may contain code executable by the data processor 108A for performing a method in the laboratory automation system that includes causing the fluid handling system to prepare a reagent mixture, such as a mixture of a sample and an antibody or antigen, used to evaluate whether there is any inaccuracy in the pipette dispensing volume based on molecular weight shift. If inaccuracy is detected, the data processor 108A may cause the fluid handling system 104 to adjust the pipette dispensing volume.

[0056] The fluid handling system 104 comprises a container handler 101, at least one fluid container 103, and a pipette arrangement 421. The container handler 101 can be any device used for handling or transporting a container. Suitable container handlers include, but are not limited to, pick-and-place devices such as a pick-and-place transfer gantry, transfer shuttles such as an extended linear reaction shuttle, or a combination of a pick-and-place transfer gantry and an extended linear reaction shuttle. The fluid container 103 can be a cuvette, tube, glass bottle, well in a pack, etc. In some embodiments, the fluid handling system 104 comprises multiple fluid containers 103, 103a, 103b, 103c, such as two containers, three containers, or four or more containers. The pipette arrangement 421 includes at least one pipette 404 (see Figures 2A and B) for dispensing measured volumes of at least one fluid into the fluid container 103. In some embodiments, the pipette arrangement 421 may include a second pipette 405 or may have three or more pipettes. Each pipette may include an ultrasonic transducer and a probe. The ultrasonic transducer applies ultrasonic vibrations to the tip of the probe to mix reagents in a reagent pack, mix contents in a fluid container, clean the probe after each use, and sense the fluid level in the fluid container. Each pipette may also include a fluid pump 414 for drawing diluents, calibrators, and reagents into the probe, and an associated valve. The fluid pump 414 may be driven by an actuator 415, such as a motor. In some embodiments, the motor is a stepper motor, which allows for precise control of the volume of fluid dispensed by the pipette.

[0057] The fluids dispensed by pipette configuration 421 can be calibrators, diagnostic reagents, diluents, or mixtures thereof, and patient samples. In some embodiments, a separate sample pipette station may be used to dispense patient samples. A separate sample pipette may also be used to dispense calibrators in some embodiments. In some embodiments, the calibrator and / or diagnostic reagents comprise specimens. Examples of specimens that may be analyzed within the automated clinical laboratory system include thyroid-stimulating hormone (TSH), prostate-specific antigen (PSA), troponin, vitamin D, and free thyroxine (T4). Diagnostic reagents may also comprise antibodies or antigens. Diluents that may be used to prepare calibrator diluents or patient samples include TRIS buffer and bovine serous albumin (BSA) buffer.

[0058] Figure 2A shows a block diagram of an automated analyzer component 400 that may be used in an automated clinical laboratory system according to one embodiment. The basic structural and functional modules of the automated analyzer component 400 may include: a sample dispensing station with a sample dispensing unit 401; a dispensing station with a main sample pipetting station 402; a bulk container dispenser 403; a fluid handling system including a pipette arrangement 421 which may include a first pair of dual reagent pipettes 404 and 405 and a second pair of dual reagent pipettes 406 and 407; a container handler which may include a first pick-and-place grasper 408, a second pick-and-place grasper 409 and a third pick-and-place grasper 410; an incubator / washing / reading station 412; a sample storage unit 411; and a reagent storage unit 413. Optionally, the sample and / or reagent storage units may be refrigerated. The sample supply unit 401 can be used to load calibrators, matrices, and reagent packs and transport them to the pipette configuration.

[0059] One or more of the pipettes 404, 405, 406, and 407 in pipette arrangement 421 may be used to prepare calibrator dilutions for use in calibrating analyzer components. Pipettes 404, 405, 406, and 407 may also be used to prepare diagnostic reagent mixtures that can be used to evaluate pipette dispensing volumes. The four pipettes 404, 405, 406, and 407 may be arranged as two double pipettes and may be independent of each other. Each of the four pipettes 404, 405, 406, and 407 may have its own fluid pump and valve, a monitoring tower, a reaction vessel carrier, and a probe. Each of the fluid pumps may be driven by a motor, preferably an actuator such as a stepper motor. Although four pipettes 404, 405, 406, and 407 are shown, it should be understood that embodiments of the present invention may include more or fewer pipettes.

[0060] Three pick-and-place grippers 408, 409, and 410 can be used to transport samples and reaction vessels (fluid containers) between various modules of the analyzer components. The first pick-and-place gripper 408 can be used to transport fluid containers between the bulk container feeder 403 or sample storage 411 and the pipette arrangement 421. The second pick-and-place gripper 409 can be used to transport fluid containers between the pipette arrangement 421 and the incubator / washing / reading station 412. The third pick-and-place gripper 410 can be used to transport fluid containers between the incubator of the incubator / washing / reading station 412 and the washing wheel (example of a washing station). A detailed description of the configuration and function of the pick-and-place grippers 408, 409, and 410 is provided in U.S. Patent No. 7,128,874 (which is incorporated herein in whole by reference). It should be understood that embodiments of the present invention may have more or fewer pick-and-place grippers. A further detailed description of the automated analyzer components is provided in PCT published application WO2018 / 217778, which is incorporated herein by reference in its entirety.

[0061] Figure 2B shows a block diagram of an alternative embodiment of the automated analyzer component 400a, where similar numbers represent a similar structure to that shown in Figure 2A. In this embodiment, the main sample pipette station 402a may be used not only to dispense patient samples for processing but also to dispense calibrators used to prepare calibrator dilutions. In this embodiment, one or more of the pipettes 404, 405, 406, or 407 dispense reagents used to prepare calibrator dilutions, and the sample pipette station 402a dispenses the calibrator. As in the embodiment of Figure 2A, pick-and-place grippers 408, 409, and 410 may be used to transport sample and fluid containers between the various modules of the analyzer component. Assay incubation, washing, and reading steps may be performed within the incubation station 412a, washing station 412b, and reading station 412c.

[0062] Figure 3 shows an illustrative flowchart illustrating the basic operating procedure for operating a fluid handling system to prepare a calibrator dilution series. Referring to Figures 1, 2A, and 2B, the process involves the operator loading a matrix pack containing the appropriate diluents for assays requiring calibration within the automated clinical laboratory system. The operator also loads calibrator glass vials or other fluid containers containing high concentrations of the appropriate calibrator onto the rack. A calibrator may contain a single sample or multiple samples. If multiple samples are present in a calibrator requiring different diluents or reagents, the different diluents or reagents may be provided through different wells in the matrix pack. A calibrator card associated with the calibrator provides an identifier, such as a barcode, for each calibrator diluent. In some embodiments, the operator manually scans the barcode information and transmits it to the control system. In preferred embodiments, the calibration card may be mounted directly on the rack so that analyzer components can directly read the calibrator information and transmit it to the control system 108. The calibrator rack is advanced to pipette position 421, where the identifier on the calibrator glass bottle is read and the assay requiring calibration is identified. Simultaneously, the fluid containers necessary for preparing the calibration dilutions are delivered to pipette position 421 by a container handler such as a pick-and-place gripper 408.

[0063] In pipette configuration 421, the required number of diluents to generate the calibration curve are prepared by dispensing the required volume of diluent from the matrix pack into the fluid container 103 from one of pipettes 404, 405, 406, or 407. In the embodiment of Figure 2A, the required volume of high-concentration calibrator is then dispensed from one of pipettes 404, 405, 406, or 407 into the same fluid container. In the embodiment of Figure 2B, the fluid container 103 is moved from the pipette configuration to the main sample pipette station 402a, which dispenses the required volume of high-concentration calibrator into the same fluid container. If a second dilution (or higher) of the calibrator is required, it may be prepared in the second fluid container 103a in the same manner as the first dilution prepared according to each embodiment (except that the second dilution differs from the first dilution, such that the required volume of diluent dispensed into the second fluid container 103a by the pipette differs for the second dilution of the calibrator). Additional calibrator dilutions may be prepared depending on the number of calibration points required for the calibration curve for a particular assay. Each prepared dilution has a different concentration of the calibrator, resulting in a set or series of calibrator dilutions. Examples of dilution series may be 1 / 1, 1 / 2, 1 / 5, 1 / 10, 1 / 15, and 1 / 20. In embodiments, each dilution may be in the range of 1 / 1 to 1 / 200. In some embodiments, each dilution volume is sufficient to perform at least three assay repeats. One set of dilution series may be transferred to mass spectrometer 106 for calibrator measurement. The measurement from mass spectrometer 106 may be used to generate an RLU dose calibration curve. The other two dilution series may be transferred to analyzer component 102 for assay testing.

[0064] Figure 4 shows an illustrative flowchart illustrating the basic operating procedure for operating a fluid handling system to prepare a mixture of diagnostic reagents for use in evaluating pipette dispensing volumes. In the process, referring to Figures 1 and 2A, the operator loads a reagent pack into the automated clinical laboratory system. The reagent pack contains at least two diagnostic reagents, one containing a sample and the second containing an antibody or antigen. The reagent pack is transferred to the pipette arrangement 421. Simultaneously, a container handler 101, such as a pick-and-place grasper 408, provides the fluid container to the pipette arrangement 421 for dispensing the diagnostic reagents into the fluid container 103. With the reagent pack and fluid container 103 in place, a first pipette, which may be one of pipettes 404, 405, 406, or 407, uses a high-precision pump to dispense a first required volume of the sample into the fluid container 103, and a second pipette, which may be another of pipettes 404, 405, 406, or 407, uses a high-precision pump to dispense a second required volume of the antibody or antigen into the same fluid container 103. The diagnostic reagents are then allowed to undergo an immunoassay and form a mixture. The mixture can then be transferred to a mass spectrometer for evaluation.

[0065] A wide variety of mass spectrometer systems that can form part of a mass spectrometer can be used in various embodiments of automated clinical laboratory systems. A suitable mass spectrometer system includes two mass separators, which are accompanied by an ion fragmentator positioned in the ion flight path between the two mass separators. Examples of suitable mass separators, but not limited to, include quadrupoles, RF multipole systems, ion traps, time-of-flight (TOF) systems, and TOF systems combined with time-delayed ion selectors. Suitable ion fragmentators, but not limited to, those that operate on the principles of collision-induced dissociation (CID, also known as collision-assisted dissociation (CAD)), photo-induced dissociation (PID), surface-induced dissociation (SID), and post-source dissociation, through interaction with electron beams (e.g., electron-induced dissociation (BID), electron-captured dissociation (BCD)), interaction with thermal radiation (e.g., thermal / blackbody infrared dissociation (BIRD)), post-source decomposition, or combinations thereof.

[0066] Suitable mass spectrometers include, but are not limited to, those featuring one or more of the following: triple quadrupoles, quadrupole linear ion traps (e.g., 4000 Q TRAP® EC / MS / MS systems, Q TRAP® LC / MS / MS systems), quadrupole TOF (e.g., QSTAR® LC / MS / MS systems), and TOF-TOF systems.

[0067] The mass spectrometer may be equipped with a triple quadrupole mass spectrometer for selecting a parent ion and detecting its fragment daughter ions. In this embodiment, the first quadrupole selects the parent ion. The second quadrupole is maintained at a sufficiently high pressure and voltage, thereby causing multiple low-energy collisions to occur and fragment some of the parent ions. The third quadrupole is selected to transmit the selected daughter ions to the detector. In various embodiments, the triple quadrupole mass spectrometer may include an ion trap placed between the ion source and the triple quadrupole. The ion trap can be configured to collect ions (e.g., all ions, ions with a specific m / z range, etc.) and, after a packing time, transmit the selected ions to the first quadrupole by pulsing the end electrodes, allowing the selected ions to exit the ion trap. The desired packing time can be determined based on, for example, the number of ions, the charge density in the ion trap, the time between elutions of different characteristic peptides, the duty cycle, the decay rate of excited state species, the number of highly charged ions, or a combination thereof.

[0068] One or more of the quadrupoles in a triple quadrupole mass spectrometer can be configured as a linear ion trap (e.g., by adding end electrodes to provide a substantially elongated cylindrical capture volume within the quadrupole). In various embodiments, the first quadrupole selects parent ions. The second quadrupole is maintained at a sufficiently high collision gas pressure and voltage to produce multiple low-energy collisions and fragment some of the parent ions. The third quadrupole is selected to capture fragment ions and, after a packing time, transmits the selected daughter ions to the detector by pulsing the end electrodes, allowing the selected daughter ions to exit the ion trap. The desired packing time can be determined based on, for example, the number of fragment ions, the charge density in the ion trap, the time between elutions of different characteristic peptides, the duty cycle, the decay rate of excited state species, or the number of highly charged ions, or a combination thereof.

[0069] In some embodiments, the mass spectrometer may comprise two quadrupole mass separators and a TOF mass spectrometer to select a parent ion and detect its fragment daughter ions. In various embodiments, the first quadrupole selects the parent ion. The second quadrupole is maintained at a sufficiently high pressure and voltage so that multiple low-energy collisions occur and fragment some of the ions, and the TOF mass spectrometer selects the daughter ions for detection, for example, by time-gate the detector to the arrival time window of the selected daughter ions by deflecting ions that appear outside the time window of the selected daughter ions away from the detector, or by monitoring the ions over a mass range encompassing the daughter ion of interest and the extracted ion chromatogram generated by a combination thereof.

[0070] In some embodiments, the mass spectrometer may comprise two TOF mass spectrometers and an ion fragmentator (e.g., CID or SID). In various embodiments, the first TOF selects a parent ion for introduction into the ion fragmentator (e.g., by deflecting ions appearing outside the time window of the selected parent ion away from the fragmentator), and the second TOF mass spectrometer selects a daughter ion for detection by time-gateping the detector to the arrival time window of the selected daughter ion, by deflecting ions appearing outside the time window of the selected daughter ion away from the detector, or by monitoring the ion over a mass range encompassing the daughter ion of interest and the extracted ion chromatogram generated by a combination thereof. The TOF analyzer may be a linear or reflection analyzer.

[0071] The mass spectrometer may comprise a tandem MS-MS instrument comprising a first field-free drift region having a timed ion selector for selecting a parent ion of interest, a fragmentation chamber (or ion fragmentator) for producing daughter ions, and a mass separator for transmitting the selected daughter ions for detection. In various embodiments, the timed ion selector comprises a pulsed ion deflector. In various embodiments, the ion deflector may be used as a pulsed ion deflector. The mass separator may include an ion reflector. In various embodiments, the fragmentation chamber is a collision cell designed to induce ion fragmentation and delay extraction. In various embodiments, the fragmentation chamber may also serve as a delayed extraction ion source for the analysis of fragmented ions by time-of-flight mass spectrometry.

[0072] In some embodiments, ionization can be used to produce structurally specific fragment ions and Q3 MRM ions. Labeling reagents can be fully or partially incorporated into the structurally specific fragment ions. The method can provide both sensitivity and specificity for the Q3 MRM ions. In some embodiments, ionization can be used to produce a dominant neutral loss fragment ion, which is selected in Q3 and then fragmented to produce structurally specific ions. These fragment ions can then be used for identification and quantification in a procedure referred to as MSS.

[0073] Figure 5 shows a block diagram of an exemplary mass spectrometer 600 and an introduction device 601 for introducing a calibrator dilution or diagnostic reagent mixture into the mass spectrometer. In some embodiments, the introduction device 601 may be located within the analyzer component. The introduction device 601 may be coupled to the mass spectrometer 600 through a connecting tube 602. The introduction device 601 may introduce the calibrator dilution to the ion source 603 through the connecting tube 602. The ion source 603 may be controlled by an ion source power supply 604 via a signal line 605A. Ions related to the calibrator molecules generated by the ion source 603 are introduced into the mass spectrometry area 606 and mass-analyzed. The mass spectrometry area 606 is evacuated to a vacuum by a vacuum system 607. The ions thus mass-analyzed are detected by an ion detector 608. The detection signal is fed to a data processing unit 609 via a signal line 605B. The data processing unit 609 may be a separate unit or may be part of the control system described above.

[0074] Figure 6 shows a schematic diagram of a part of a mass spectrometer using the electrospray method. Figure 6 is a cross-sectional view showing the structure of the introduction device 619 coupled to the electrospray ion source. The calibrator dilution or reagent mixture supplied from the introduction device 619 is introduced into the capillary tube 621 through the connecting tube 622 and connection 630 for spraying. By applying a voltage of the magnitude of kV between the spray capillary tube 621 and the counter electrode 632, small charged droplets of the calibrator dilution or reagent mixture are sprayed conically from the end of the spray capillary tube, i.e., the so-called electrospray phenomenon occurs. In the electrospray method, an output 623 for the spray gas is provided, and a gas such as nitrogen gas is injected around the spray capillary tube 621, thereby accelerating the vaporization of the small charged droplets. Furthermore, a gas such as nitrogen gas is blown towards the small charged droplets generated from the outlet 624 to vaporize the gas supplied to the counter electrode 632 and accelerate the vaporization of the small charged droplets. The generated ions are introduced into the vacuum section 626 through the ion sampling opening 625 and subjected to mass spectrometry under high vacuum by the mass spectrometry region 626.

[0075] Figure 7 shows the structure of an ion detector. The structure shown in Figure 7 can be used to improve the signal-to-noise ratio (SIN) in a mass spectrometer. An ion deflection electrode 646 can be provided in the rear portion of the mass spectrometry region 648 for mass separation under a high-frequency electric field to deflect mass-separated ions. The deflected ions are accelerated at a voltage on the scale of kV and collide with a dynode 657, producing secondary electrons. The secondary electrons are emitted from the secondary electron-producing dynode 657 upon which the ions collide. The emitted secondary electrons are detected by an electron detector 658, such as an electron multiplier tube. The structure shown in Figure 7 prevents uncharged neutral molecules, charged droplets, or uncharged droplets from being detected as signals by the ion detector 648, so as to achieve an improved S / N ratio.

[0076] Figure 8 shows a flowchart illustrating the process steps for generating a calibration curve according to an exemplary embodiment. It can be referenced with respect to the automated analyzer components of Figures 2A and 2B. In step 802, the matrix pack and calibrator are loaded into the presentation unit 401 within the automated analyzer component 400. In step 804, one of the pipettes 404, 405, 406, or 407 delivers the required volume of diluent into a fluid container 103 provided by the bulk container feeder 403. In step 806, the same pipette or another of the pipettes 404, 405, 406, or 407 in the embodiment of the automated analyzer component in Figure 2A delivers the required volume of calibrator into the same fluid container 103. In the embodiment of Figure 2B, dispensing the calibrator 806 is performed by the sample pipette station 402a. Steps 804 and 806 are repeated for each calibrator diluent to be prepared. In step 808, a series of calibrator dilutions are transferred to the mass spectrometer 106 for measurement. In step 810, the measurements from the mass spectrometer 106 are used to automatically generate an RLU dose calibration curve. The actual sample can then be analyzed in the analyzer component 102 based on the calibration curve generated in step 810, so that quantitative values ​​of the specimen in the sample can be obtained.

[0077] It should be understood that additional process steps may need to be performed depending on the specific assay to be calibrated. For example, magnetic beads or particles may be added, and incubation, separation, and / or washing steps may be performed. Other processing steps may include immunopurification processing steps. In the immunopurification process, the sample is captured by the antibody, and any unbound molecules are washed away in the washing process. In the subsequent elution step, the sample is then released from the antibody using a buffer and eluent. The eluent containing the "purified" target can be characterized as a processed sample aliquot, which is then collected and analyzed by mass spectrometry. Other processing steps may include protein cleavage processing and SISCAPA-type processing steps. Rather than directly measuring undamaged protein by mass spectrometry, SISCAPA utilizes proteolysis (e.g., using the enzyme trypsin) to cleave the sample protein into smaller peptides that are ideally suitable for quantification by mass spectrometry. By selecting a target peptide whose configuration occurs only in the target protein (a so-called "proteotype" peptide), the target peptide can serve as a direct quantitative substitute for the target protein.

[0078] Figure 9 shows a flowchart illustrating process steps for preparing a diagnostic reagent mixture that can be used to evaluate the dispensing volume of a selected pipette according to one embodiment of the present invention. Figures 1 and 2A can be referenced. In step 902, a reagent pack comprising a first diagnostic reagent containing a sample and a second diagnostic reagent comprising an antibody or antigen is loaded into a presentation unit 401 in an automated analyzer 400. In step 904, one of pipettes 404, 405, 406, or 407 delivers a required volume of the first diagnostic reagent into a fluid container 103 supplied by a bulk container feeder 403. In step 906, another pipette 404, 405, 406, or 407 delivers a required volume of the second diagnostic reagent into the same fluid container 103. The mixture of diagnostic reagents in the fluid container 103 can be mixed using any preferred mixing process. In step 908, a fluid container 103 containing a mixture of reagents is incubated to form sample / antibody or sample / antigen complexes. In step 910, the sample / antibody or sample / antigen complexes are transferred to a mass spectrometer 106, and the number of sample / antibody or sample / antigen complexes is quantified by molecular weight. In step 912, a bias is calculated based on the ratio of the measured signal from the mass spectrometer to the expected signal. If the measured signal does not meet a certain threshold, such as 90% of the expected signal, an inaccuracy in the dispensing volume is detected in the selected pipette being evaluated. If an inaccuracy is detected, the dispensing volume of the selected pipette requests adjustment. In step 914, a coefficient of 1 / bias value is applied to an actuator for a dispensing pump associated with the target pipette, and the actuator is adjusted as necessary to compensate for the detected inaccuracy in the dispensing volume. In some embodiments, the actuator 415 may be a stepper motor.

[0079] The clinical laboratory automation system 100, in various embodiments, can be used to measure or determine the presence of various specimens, such as hormones, illicit drugs, and tumor markers, in one or more samples. With respect to many of these specimens, the clinical laboratory automation system 100 can provide automated calibration using a single calibrator to prepare one or more calibrator dilutions used to generate calibration curves for quantitative measurement of the target specimen.

[0080] Additional examples are provided below.

[0081] (Examples)

[0082] (Example 1: Dilution of a single thyroid-stimulating hormone calibrator)

[0083] Measuring thyroid-stimulating hormone (TSH) is useful in assessing thyroid function and monitoring patients undergoing thyroid replacement therapy. TSH is part of the hypothalamic-pituitary-thyroid axis, which regulates the body's metabolism. The hypothalamus secretes thyroid-stimulating hormone-releasing hormone (TRH), which stimulates the pituitary gland to secrete TSH. TSH triggers the release of thyroid hormones T3 (triiodothyronine) and T4 (thyroxine), which regulate metabolic function within cells. Excessive amounts of T3 or T4 circulating lead to a process where TRH production stops, controlled by a negative feedback loop.

[0084] Quantitative analysis of TSH in patient samples requires preliminary calibration of the instrument response of the instruments and equipment used to detect the amount of TSH in the sample being analyzed. Calibration is typically performed using a known concentration of the target sample present in the sample to be analyzed. The known concentration can be used to construct a calibration curve that plots the mathematical relationship between the measured response and the known sample concentration on a graph. One difficulty that may arise is that the calibration signal (i.e., relative light units) may degrade over time, while the assigned dose value remains the same. This difficulty is illustrated in Figure 10A.

[0085] A new calibration curve can be prepared by the following method: A single calibrator, TSH, is loaded onto a fluid handling system integrated with the immunoassay analyzer. A series of dilutions are produced by first dispensing the diluent, and then dispensing the calibrator. The diluted TSH calibrator is then sent to a mass spectrometer. The concentration of each calibrator dilution is quantified on the mass spectrometer, and the measured concentrations assigned to each calibrator dilution are provided on the immunoassay analyzer. The new calibration curve, shown in Figure 10B, is then plotted on the immunoassay analyzer.

[0086] (Example 2: Master Calibration Curve Adjustment)

[0087] In some embodiments, a manufacturer may generate a master calibration curve for an automated clinical laboratory system at its manufacturing facility. The manufacturer provides the customer with specific calibration information, such as a barcode or 2D code accompanying the reagent package, and one or two calibrators. The user can then test the calibrators on their analyzer and subsequently adjust the RLU dose calibration curve in-situ based on the values ​​generated with respect to the calibrators. Figure 10C illustrates this adjustment.

[0088] (Example 3: Adjustment of pipette dispensing volume)

[0089] A fluid handler loads two diagnostic reagents onto an immunoassay analyzer. The first diagnostic reagent contains the sample, and the second diagnostic reagent contains the antibody. A mixture of the two diagnostic reagents is produced, in which the amount of one reagent is significantly greater than that of the other. The mixture is then sent to a mass spectrometer. The mass spectrometer quantifies the number of sample / antigen complexes by molecular weight shift (Figure 11), and a bias is calculated from the measured signal / expected signal. If the measurement is not within a predetermined threshold, a coefficient (=1 / bias) is applied to the target pipette motor step.

[0090] (Example 4: Testosterone)

[0091] The Access Testosterone Assay Kit (commercially available from Beckman Coulter, Inc. (Brea, CA)) can be used for initial testing of testosterone in biological samples. The assay can be performed on an immunoassay analyzer in a sample processing system. The Access Testosterone Assay is a competitive binding immunoenzyme assay using paramagnetic particles coated with mouse monoclonal anti-testosterone antibody, testosterone alkaline phosphatase conjugate, and goat anti-mouse polyclonal antibody. Testosterone in the sample is released from the carrier protein and competes with the testosterone alkaline phosphatase conjugate for site binding with respect to a limited amount of specific anti-testosterone monoclonal antibody. The resulting antigen / antibody complex is then bound to the solid phase by the capture antibody. After incubation in a reaction vessel, the material bound to the solid phase is held in a magnetic field, while the unbound material is washed away. Next, the chemiluminescent substrate Lumi-Phos*530 is added to the container, and the light generated by the reaction is measured using a luminance meter. Light production is inversely proportional to the concentration of testosterone in the sample. The amount of sample in the sample is determined from a stored multi-point calibration curve generated from mass spectrometer measurements of a calibrator dilution series prepared from a high-concentration calibrator in the Access Testosterone Kit.

[0092] (Example 5: Amphetamine)

[0093] An automated clinical laboratory system according to one embodiment of the present disclosure can be used to test for drugs in misuse. One exemplary drug of the misuse type is amphetamine. Amphetamines are central nervous system stimulants that produce a sense of alertness, vigilance, increased energy, reduced hunger, and overall well-being.

[0094] Amphetamines appear in the urine within 3 hours of any type of administration and can be detected over a period of 24–48 hours from the last dose by the Emit® II and amphetamine assay (commercially available from Beckman Coulter Inc. (Brea, CA)). The Emit® II and amphetamine assay is a homogeneous enzyme immunoassay. The assay is based on competition between the drug in the sample and the drug labeled with the enzyme glucose-6-phosphate dehydrogenase (G6PDH) with respect to the antibody binding site. Enzyme activity decreases upon binding to the antibody, and therefore the drug concentration in the sample can be measured in terms of enzyme activity. The active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH, resulting in a spectrophotometrically measured change in absorbance. Endogenous serous G6PDH does not interfere because the coenzyme NAD functions only with the enzyme of the bacterium (Leuconostoc mesenteroides) employed in the assay.

[0095] The reagents used in the test may include dextroamphetamine (61 μg / mL) and d-methamphetamine (10 μg / mL), glucose-6-phosphate (5.5 mM), nicotinamide adenine dinucleotide (3.5 mM), bovine serous albumin, amphetamine labeled with bacterial G6PDH (0.72 U / mL), TRIS buffer, preservatives, and mouse monoclonal antibodies against stabilizers. The processed sample is analyzed and compared to an assay threshold generated from mass spectrometer measurements of calibrator dilutions prepared from the calibrator in the assay kit according to the exemplary embodiment.

[0096] (Example 6: Heart disease and stroke)

[0097] In some embodiments, automated clinical laboratory systems can be used to detect the risk of heart disease or stroke. Many forms of cardiovascular disease begin with atherosclerosis, a condition in which arteries harden and narrow due to the accumulation of plaque around the arterial walls. Plaque, consisting of cholesterol, fatty substances, cellular waste products, calcium, and fibrin, can partially or completely block the flow of blood through arteries in the heart, brain, pelvis, lower extremities, arms, or kidneys. This blockage can develop into serious conditions such as coronary heart disease, chest pain, carotid artery disease, peripheral artery disease (PAD), and chronic kidney disease. Worse still, if fragments of plaque rupture or blood clots (thrombi) form on the surface of the plaque, a heart attack or stroke may result.

[0098] Several lipoprotein markers are good biomarkers for cardiac disease, and stroke can be measured from body fluid samples collected from patients, e.g., blood, plasma, and serous fluid, using mass spectrometry. These markers include type B natriuretic peptide (BNP), proBNP (an inactive prohormone that produces BNP), human C-reactive protein (hs-CRP), and pregnancy-associated plasma protein A (PAPP-A). Many of these natriuretic peptides can help determine the risk of plaque progression and stroke development. Other markers include HDLp (high-density lipoprotein) ratio, lipofolin / cholesterol ratio, lipid / lipofolin ratio, LDL cholesterol levels, HDLp and apolipoprotein levels, lipofolin and LTP ratio, sphingolipids, omega-3 index, and triglycerides for ST2 levels, which can be assayed using mass spectrometry or analyzer components of automated systems. Quantitative measurements can be determined based on calibration curves generated from mass spectrometer measurements of calibrator dilutions. The measurements can be compared to reference ranges according to pre-established rules for determining the risk of heart disease or stroke.

[0099] If a small portion of the tumor markers are indicated as positive according to the mass spectrometer data, the control system in the sample processing system instructs the sample preparation module in the immunoassay analyzer to prepare and process a second aliquot of the sample. The immunoassay analyzer then detects the portion of the tumor markers using a multiplex fluorescence-based sandwich immunoassay. The assay may involve adding a primary antibody specific to each tumor marker in the portion, and a detection antibody conjugated to a fluorescent material and capable of recognizing each of the primary antibodies, to the sample aliquot. The fluorescent materials have different excitation and emission wavelengths, and therefore the fluorescence signals from the detection antibodies will not interfere with each other. The fluorescence signal from each detection antibody is measured, and it represents the amount of each corresponding tumor marker in the sample. The results for the tumor markers determined to be positive by the immunoassay analyzer are then reported.

[0100] The above description is illustrative and not restrictive. Many modifications of the invention will become apparent to those skilled in the art upon closer examination of this disclosure. The scope of the invention should therefore not be determined by reference to the above description, but rather by reference to the pending claims, together with their entire scope or equivalents.

[0101] All patents, patent applications, publications, and descriptions mentioned above are incorporated herein by reference as a whole.

Claims

1. A clinical laboratory automation system, wherein the clinical laboratory automation system is A fluid handling system comprising a container handler, at least one fluid container, and a pipette arrangement, wherein the fluid handling system is configured to dispense at least a first fluid and a second fluid into the at least one fluid container, the first fluid and the second fluid being different fluids selected from the group consisting of calibrators, diluents, or mixtures thereof, and the fluid handling system is configured to produce a dilution series of the calibrator, the dilution series comprising at least one diluent of the calibrator, Analysis components, A mass spectrometer configured to evaluate the properties of at least one dilution of the calibrator, thereby producing a corresponding set of values ​​by using the dilution series of the calibrator. Equipped with, A clinical laboratory automation system in which the analytical component is calibrated at least partially using a corresponding set of the aforementioned values.

2. The clinical laboratory automation system according to claim 1, wherein the analytical component includes an immunoassay analyzer.

3. The automated clinical laboratory system according to claim 1, wherein the analytical component includes a clinical chemistry analyzer, a protein chemistry analyzer, a hematology analyzer, or a urinalysis analyzer.

4. The clinical laboratory automation system according to claim 1, further comprising a sample pipette station.

5. The automated clinical laboratory system according to any one of claims 1 to 4, wherein the pipette arrangement comprises a pump, the pump being driven by an actuator.

6. The automated clinical laboratory system according to claim 1, wherein the calibrator comprises at least one type of sample or antibody.

7. The automated clinical laboratory system according to claim 4, wherein the pipette arrangement further comprises at least a first pipette, the first pipette being configured to dispense at least one required volume of a fluid.

8. The automated clinical laboratory system according to claim 7, wherein the first fluid is the diluent and the second fluid is the calibrator.

9. The clinical laboratory automation system according to claim 8, wherein the characteristic to be evaluated is the concentration of the at least one dilution of the calibrator from the set of dilution series of the calibrator.

10. The automated clinical laboratory system according to claim 9, wherein an RLU dose conversion curve is generated from a corresponding set of the values, and the analytical component is calibrated at least partially using the RLU dose conversion curve.

11. The automated clinical laboratory system according to claim 1, wherein at least a portion of the fluid handling system is integrated with the mass spectrometer.

12. The clinical laboratory automation system according to claim 1, further comprising a control system configured to control the fluid handling system, the analytical components, and / or the mass spectrometer.

13. A method for calibrating an analytical component, wherein the method is: The steps include providing an automated clinical laboratory system, The aforementioned automated clinical laboratory system is (a) A fluid handling system comprising a container handler, at least one fluid container, and a pipette arrangement, wherein the pipette arrangement comprises at least a first pipette and at least a second pipette, the first pipette configured to dispense a first required volume of a first fluid into the fluid container, the second pipette configured to dispense a second required volume of a second fluid into the same fluid container, the first fluid and the second fluid being different fluids selected from the group consisting of calibrators, diluents, or mixtures thereof, the fluid handling system being configured to produce a dilution series of the calibrator, the dilution series comprising at least one diluent of the calibrator, (b) an immunoassay analyzer or clinical chemistry analyzer, (c) A mass spectrometer (106) configured to evaluate the properties of at least one dilution of the calibrator, thereby producing a corresponding set of values ​​by using the dilution series of the calibrator. Equipped with, The method further comprises a clinical laboratory automation system configured to calibrate the immunoassay analyzer or clinical chemistry analyzer using, at least in part, a corresponding set of the aforementioned values.

14. A method for calibrating an analytical component, wherein the method is: To provide an automated clinical laboratory system comprising a fluid handling system, analytical components, and a mass spectrometer, wherein the fluid handling system includes a container handler, at least a first fluid container, and a pipette arrangement, and the pipette arrangement comprises at least a first pipette and a second pipette. To provide a calibrator, To provide a diluent, A dilution series comprising at least one dilution of the calibrator, Using the aforementioned pipette arrangement, the required first volume of the diluent is dispensed into the first fluid container (103), Using the aforementioned pipette arrangement, the required first volume of the calibration agent is dispensed into the first fluid container (103). By producing, The mass spectrometer is used to evaluate the properties of at least one dilution of the calibrator, thereby generating corresponding sets of values ​​by using the dilution series. Calibrating the analysis component using, at least partially, the corresponding set of the aforementioned values ​​and Methods that include...

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