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WO2015058158A1 - Procédé pour la détection d'une association lp-pla2 spécifique aux lipoprotéines - Google Patents

Procédé pour la détection d'une association lp-pla2 spécifique aux lipoprotéines Download PDF

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Publication number
WO2015058158A1
WO2015058158A1 PCT/US2014/061259 US2014061259W WO2015058158A1 WO 2015058158 A1 WO2015058158 A1 WO 2015058158A1 US 2014061259 W US2014061259 W US 2014061259W WO 2015058158 A1 WO2015058158 A1 WO 2015058158A1
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Prior art keywords
pla2
solid
lipoprotein
probing
phase support
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Inventor
Yan Liu
Thomas D. Schaal
Xiaozhu Duan
Shaoqui ZHUO
Leonard MENDIOLA
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Diadexus Inc
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Diadexus Inc
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/44—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving esterase
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/916—Hydrolases (3) acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
    • G01N2333/918—Carboxylic ester hydrolases (3.1.1)

Definitions

  • compositions, kits and methods related to detection for detection of specified lipoproteins expressing Lp-PLA2 in a sample are described herein.
  • Lp-PLA 2 lipoprotein-associated phospholipase A2
  • Lp-PLA 2 lipoprotein-associated phospholipase A2
  • HDL high-density lipoprotein
  • Described herein are methods and systems capable of achieving adequate signal in an assay (e.g., a hybrid ELISA assay) using a PLAC antibody (Lp-PLA 2 antibody). These methods and systems may allow determination of lipoprotein specific Lp-PLA 2 levels in patient samples and/or the determination of relative levels of Lp-PLA 2 associated with different lipoproteins.
  • an assay e.g., a hybrid ELISA assay
  • PLAC antibody Lp-PLA 2 antibody
  • an antibody recognizing an Lp-PLA 2 antigen was used as the capture antibody, which may be bound to a substrate, and one or a set of individual antibody conjugates recognizing various lipoprotein subspecies (for example, Apo-Al, Apo-B and apo[a]) are used as a detection antibody.
  • the subspecies may be assayed in parallel, either separately or concurrently.
  • Antibodies may be conjugated to different reporters (e.g., fluorophore).
  • the methods and systems implementing these methods includes the use of a Lp-PLA2 antibody as the capture antibody to initially capture lipoproteins associated with Lp- PLA2, then screening the resulting enriched population for specific lipoprotein subclasses (e.g., LDL, HDL, Lp(a)).
  • This approach is the inverse of prior art systems, in which capture antibodies were directed to lipoproteins which were then screened to determine association with Lp-PLA2.
  • Lp-PLA2 Lipoprotein-associated phospholipase A2
  • a method may include: enriching a patient sample for lipoproteins expressing Lp- PLA2; and detecting a subclass of lipoprotein particles from the enriched sample.
  • Enrichment of the patient sample typically refers to enrichment by binding of Lp-PLA2 within the patient sample to a solid-phase material to which a capture molecule for Lp-PLA2 has been linked.
  • a method of detecting lipoprotein particles having an associated Lipoprotein- associated phospholipase A2 (Lp-PLA2) molecule from a patient sample may include, for example: exposing a solid-phase support material in which a capture molecule that specifically binds Lp-PLA2 have been immobilized to the patient sample; and probing the solid-phase support material with a detection molecule specific a subclass of lipoprotein particles.
  • any of these methods may also be methods of detecting a patient's relative association of Lipoprotein-associated phospholipase A2 (Lp-PLA2) with one or more lipoproteins.
  • the method may include: exposing a solid-phase support material to a patient sample, wherein a capture molecule that specifically binds Lp-PLA2 has been immobilized on the one or more solid phase supports; probing the solid-phase support material with a detection molecule specific to Lp-PLA2; and probing the solid-phase support material with a detection molecule specific to a subclass of lipoprotein particles.
  • the capture molecule that specifically binds Lp-PLA2 may be, for example, an antibody or antibody fragment directed to Lp-PLA2.
  • any of these methods may include incubating the solid-phase support in the patient sample for greater than 10 minutes before washing the solid-phase support material, to enrich the sample.
  • probing may comprise probing with a labeled antibody (and/or antibody fragment, e.g., Fab, etc.).
  • Probing the solid-phase support material with a detection molecule specific to Lp-PLA2 and probing the solid-phase support material with a detection molecule specific to a subclass of lipoprotein particles may comprise probing parallel samples including the solid-phase support material.
  • probing the solid-phase support material with a detection molecule specific to Lp-PLA2 and probing the solid-phase support material with a detection molecule specific to a subclass of lipoprotein particles may comprise probing the same sample including the solid-phase support material.
  • Probing may comprises probing with an antibody specific to one or more of: Apo-Al, ApoB-100/-48, and Apo(a).
  • the methods include probing the solid phase support with an antibody to a second (or more) subclass of lipoprotein particles, and may also include calculating a ratio from the detected levels of the different subclasses of lipoprotein particles. [00015] In general, any of these methods may also include calculating a ratio from detected levels of the lipoprotein particles and the Lp-PLA2 determined by the probing steps.
  • kits for detecting lipoprotein particles having an associated Lipoprotein-associated phospholipase A2 (Lp-PLA2) molecule from a patient sample may include: a solid-phase support material to which a protein that specifically binds Lp-PLA2 has been immobilized; a wash solution; a buffer solution; a first detection antibody specific to a first subclass of lipoprotein particles; and a second detection antibody specific to a second subclass of lipoprotein particles.
  • FIG. 1 A schematically illustrates prior-art assays to determine levels of lipoproteins associated with Lp-PLA 2 (described, for example, in figure 1C of Ami et al (2012)).
  • FIG. IB schematically illustrates a replicated version of a prior art assays to determine levels of lipoproteins associated with Lp-PLA 2 .
  • FIG. 2 shows the results of an ELISA assay using a commercially available
  • FIG. 3 shows the results of an ELISA assay using the commercially available Mercodia Lp(a) as a capture antibody and 4B4 (Lp-PLA2) antibody as a detection antibody from a serum sample.
  • FIG. 4 shows the results of an ELISA assay using 2C 10 as a capture antibody and Mercodia Lp(a) as a detection antibody from a serum sample.
  • FIG. 5 schematically illustrates the configuration for detection of lipoprotein-associated Lp-PLA2 described herein.
  • FIG. 6 shows the results of an ELISA assay using an Lp-PLA2 capture antibody (2C10) with a variety of different detection antibodies targeting specific subclasses of lipoproteins, including LDL, Lp(a), and HDL from a serum sample.
  • FIGS. 7A-7C show graphs analyzing the Lp-PLA2 (e.g., 2C10 antibody) capture and enrichment method for various lipoprotein detection antibody configurations.
  • Lp-PLA2 e.g., 2C10 antibody
  • FIG. 8A is a schematic illustration of one variation of a method for detecting subclasses of lipoproteins (such as HDL, LDL, etc.) that associate with Lp-PLA2 form a patient sample.
  • subclasses of lipoproteins such as HDL, LDL, etc.
  • FIG. 8B is a schematic illustration of different methods (method 1, method 2) for detecting subclasses of lipoproteins that associate with Lp-PLA2 from a patient sample.
  • FIG. 9 is a comparison of the two methods for detecting subclasses of lipoproteins illustrated in FIGS. 8B using patient samples. In this example, method 2 resulted in a slightly higher signal, which may represent an enrichment of the Lp-PLA2 and associated lipoprotein.
  • FIGS. 10A and 10B illustrate the results of a side-by-side comparison of an Lp-PLA2 capture method (similar to that shown in FIGS. 8A and 8B, also referred to as a "Hybrid ELISA" assay) and a prior art Lp-PLA2 detection assay (the commercially available PLAC assay described herein).
  • LpPLA2-liproprotein e.g., ApoB
  • the results indicate the detection of Lp-PLA2 and the ability to correlate the results with the patient's physiology.
  • kits for detecting lipoproteins, e.g., lipoprotein sub-classes, associated with Lp-PLA2 from a patient sample (material).
  • These systems and methods may initially enrich for Lp-PLA2 containing lipoproteins from a patient material (e.g., blood, blood fraction, bodily fluid, etc.), and then probe the resulting enriched sample to identify one or more (e.g., two, three, four, five, six, etc.) types of lipoproteins, including (but not limited to) HDL, LDL, and Lp(a).
  • the methods and systems implementing them may normalize the amount of starting antigen bound in a first incubation. For example, in the first incubation, one may want to bind a significant amount (e.g. virtually all) of the antigen present in the sample so that the sensitivity of the assay is governed significantly (e.g. solely) by the concentration of the detection antibody.
  • the configuration described herein may allow that scenario to be achieved, by using the Lp-PLA2 antibody as a capture antibody to enrich the antigen present.
  • the approach described herein may normalize the assay by capturing Lp- PLA 2 in a first incubation and allowing the lipoprotein-specific detection antibody concentration to be adjusted in a second incubation, thereby potentially allowing calibration to some established concentration of the individual lipoprotein.
  • configurations described herein may be used as part of a kit including standard reagents and processes to provide novel assays for determining, for example, the distribution of Lp-PLA2 between one or more lipoprotein fractions within a tissue (e.g., blood) sample.
  • novel and surprisingly effective method and systems for detecting the association (including in some variations, one or more of the level, amount, percent, degree, fraction, or some combination thereof) of association of Lp-PLA2 with one or more lipoprotein (e.g., LDL, HDL, etc.) and/or with lipoprotein-associated proteins (including sub-classes of lipoproteins).
  • lipoprotein e.g., LDL, HDL, etc.
  • lipoprotein-associated proteins including sub-classes of lipoproteins
  • Lp-PLA 2 sub-stoichiometric relationship of Lp-PLA 2 with the individual lipoprotein is low (e.g., between 1 : 100 and 1 : 10,000). Further, immobilized lipoprotein-specific coating antibody may become saturated with non-Lp-PLA 2 -containing lipoprotein complexes, resulting in low signal generation with the a-Lp-PLA 2 detection antibody.
  • the methods and assays (including systems) described herein may provide a solution that is not effected by the stoichiometric relationship of Lp-PLA2 and various lipoproteins. Instead, these assays may begin by enriching for lipoproteins displaying Lp-PLA2 and then probe to determine the presence, and/or distribution of different lipoprotein sub-classes.
  • a capture molecule e.g., an antibody, e.g., 2C10 antibody, diaDexus
  • Lp-PLA 2 antigen e.g., an Lp-PLA 2 antigen
  • a set of individual antibody conjugates recognizing various lipoprotein subspecies such as, Apo-Al, Apo-B and apo[a]; Table 1
  • Table 1 Relationship between Apolipoproteins:Lipoprotein Particles
  • lipoproteins containing Lp-PLA2 By enriching for lipoproteins containing Lp-PLA2, an enhanced signal may be achieved.
  • the methods described herein may enrich in a manner that is independent of the subclass of lipoprotein, allowing determination of normalized and/or relative distributions of various subclasses of lipoproteins containing Lp-PLA2.
  • the methods described herein may allow the determination of one or more ratios of one or more sub-classes of lipoproteins containing Lp-PLA2.
  • the methods described herein may allow determination of a ratio of HDL containing Lp-PLA2 to LDL containing Lp-PLA2 (or LDL containing Lp-PLA2 to HDL containing Lp-PLA2), HDL containing Lp-PLA2to Lp(a) containing Lp-PLA2 (or Lp(a) containing Lp-PLA2to HDL containing Lp-PLA2), or LDL containing Lp-PLA2 to Lp(a) containing Lp-PLA2 (or Lp(a) containing Lp-PLA2 to LDL containing Lp-PLA2) or a relative fraction of LP A containing Lp-PLA2:Lp(a) containing Lp-PLA2:HDL containing Lp-PLA2.
  • the measure of lipoprotein containing Lp-PLA2 may be normalized by a normalization factor (e.g., based on total capture Lp-PLA2).
  • a normalization factor e.g., based on total capture Lp-PLA2.
  • the amount of sample applied may be in excess, so that biding to lipoproteins is not limiting.
  • the same samples, or parallel samples may be used to determine the distribution of different associated lipoproteins (and/or sub-classes of lipoproteins) from the enriched Lp-PLA2 in a sample.
  • different detection signals e.g., different flourophores, etc.
  • kits are not limited by the Lp-PLA 2 bound lipoprotein concentration, since associated lipoprotein particles are captured first, followed by detection with the lipoprotein-specific antibody on the Lp-PLA 2 -enriched population of lipoproteins now immobilized on the solid phase support.
  • FIG. 5 is a schematic of one variation of this method.
  • apolipoprotein ApoB is not necessarily absolutely specific for LDL, see Table 1 [00042]
  • a hybrid ELISAs similar to those described by Arai et al. (2012) in which the Hpoprotein-specific antibody was utilized as the coating antibody and a a-Lp-PLA 2 detection antibody; conjugate, resulted in low signal generation.
  • results using the Mercodia capture antibody-coated plate from their commercially available Lp(a) kit were examined.
  • kits oc-Apo(a) coated plates coupled with parallel detection performed using both the kit detection antibodyxonjugate (a- Apo[a] -HRP) and the ct-Lp-PLA 2 detection antibody conjugate (4B4-HRP).
  • kit detection antibodyxonjugate a- Apo[a] -HRP
  • 4B4-HRP the kit detection antibodyxonjugate
  • FIG. 2 the complete Mercodia Lp(a) ELISA kit gave adequate signal (control, FIG. 2)
  • parallel detection with the Lp-PLA 2 detection antibodyxonjugate yielded only very weak signal generation (FIG. 3).
  • FIG. 3 Conceptually similar configurations in which a Hpoprotein-specific antibody was used as the capture antibody in combination with the 4B4-HRP detection antibody gave weak signal generation (data not shown).
  • Configurations described herein may use a different assay configuration in which a plate or other support substrate (e.g. a 2C10-coated plate such as a standard Gen-3 PLAC ELISA assay) may be used to capture Lp-PLA 2 antigen, which is followed by detection with lipoprotein- specific polyclonal antibodies directed against a lipoprotein (e.g., ApoB-100, Apo(a) and Apo- A l ). Any appropriate antibody directed to a lipoprotein (monoclonal, polyclonal, fragment, etc.), or to another constituent indicative or characteristic of a lipoprotein particle to be detected, may be used.
  • a plate or other support substrate e.g. a 2C10-coated plate such as a standard Gen-3 PLAC ELISA assay
  • lipoprotein-specific polyclonal antibodies directed against a lipoprotein e.g., ApoB-100, Apo(a) and Apo- A l
  • PLAC assays were performed on undiluted human serum samples using otherwise standard PLAC assay conditions (i.e., the standard conjugate diluent, detection antibody incubation times, wash buffers, etc.).
  • Standard PLAC assay conditions are known based on, for example, diaDexus' published PLAC assay.
  • a standard (low) detergent concentration and (high) salt concentration utilized in the PLAC assay may be beneficial to avoid dissociation of Lp-PLA 2 from the different lipoproteins.
  • Good signal intensity was obtained using all three lipoprotein-specific detection antibodies in these studies (FIG. 6).
  • different human serum samples demonstrated different OD levels within a given analyte measurement (FIG. 7) and/or different relative OD levels between different analyte measurements on percentage basis (Table 3).
  • Lp-PLA2 lipoprotein-associated phospholipase A2
  • Apolipoprotein B is the main protein component of LDL and ApoB is regarded as a negative factor in cardiovascular disease.
  • ApoB- 100 is normally 98% of the ApoB in plasma and at a
  • an LpPLA2-Apolipoprotein hybrid Assay as described herein may be used to quantify the Apolipoprotein associated LpPLA2 (or LpPLA2 associated
  • Apolipoprotein using a capture molecule (such as an antibody or antibody fragment) that recognizes an Lp-PLA2 antigen (e.g., the diaDexus 2C10 antibody) to capture Lp-PLA2 in a manner that enriches for the capture from the sample, while preserving the association with lipoproteins, and then probing for the lipoproteins or associated markers from the enriched sample.
  • a capture molecule such as an antibody or antibody fragment
  • an Lp-PLA2 antigen e.g., the diaDexus 2C10 antibody
  • Apo-AI , Apo-B and Lp(a) detection antibodies may be used. In such an enriched system, it may be difficult to quantify the concentration of Lp-PLA2/Apolipoprotein complexes.
  • FIG. 8A schematically illustrates one example of a method of detecting lipoprotein particles having an associated Lipoprotein-associated phospholipase A2 (Lp-PLA2) molecule from a patient sample.
  • Lp-PLA2 capture antibody (2C10) is bound to a substrate and is exposed to a biological sample (e.g., blood, serum, plasma, etc.). This step may be performed over a sufficient time to drive binding (e.g., 10 min or longer, e.g., 15 min or longer, 20 min or longer, 30 min or longer, 40 min or longer, 1 hr.
  • the enriched sample (solid phase) may be probed to detect one or more class/subclass of lipoprotein particles.
  • the enriched samples are probed for an ApoB protein using an antibody directed to ApoB. Detection may be made, and in some cases quantified, using an indicator such as HRP (e.g., by using a biotinylated detection antibody and a streptavidin-HRP molecule).
  • HRP e.g., by using a biotinylated detection antibody and a streptavidin-HRP molecule.
  • this method identified a direct interaction between ApoB and Lp-PLA2 in the LDL (and for these patients, only in LDL).
  • FIG. 8B illustrates two alternative methods.
  • method 1 an initial incubation to enrich the Lp-PLA2 from the sample by binding to a solid phase substrate (e.g., dish, beads, etc.) onto which anti-Lp-PLA2 was bound at relatively high density.
  • a lipoprotein detection molecule e.g., a biotinylated anti-ApoB antibody
  • the initial step of enriching for Lp-PLA2 on the solid phase substrate may be performed for a longer period (e.g., in this example, 2 hours), followed by a wash. Thereafter, the enriched sample may be probed with the lipoprotein detection moiety, and detected.
  • the method of enriching for LpPLA2 to the solid phase support/bound LpPLA2 and washing prior to detecting any affiliated molecules (e.g., lipoproteins) enhanced the resulting signals.
  • the enrichment over >10 min may allow the LpPLA2 capture molecules (bound anti-Lp-PLA2) to capture more Lp-PLA2 (and therefore more Lp-PLA2/ApoB or other lipoproteins complex).
  • the methods described herein may be used to analyze the relative amount of Lp-PLA2 bound to one or more lipoprotein.
  • the method may identify Lp-PLA2 using an Lp-PLA2 capture molecule and an Lp-PLA2 detection molecule, and in parallel (from parallel or concurrent samples) may identify one or more lipoproteins associated with the Lp-PLA2, as shown in FIGS. 10A and 10B, using an Lp-PLA2 capture molecule and a I ipoprote in-associated detection molecule (e.g., anti-ApoB).
  • the relative amounts are determined even in the absence of one or more calibrators for the absolute amounts.
  • FIG. 10B shows a comparison (shown as a percentage of lipoprotein- associated [ApoB] Lp-PLA2 to Lp-PLA2).
  • Comparisons of the distribution of Lp-PLA2 among different lipoproteins over time may be used to guide treatment.
  • within-patient monitoring of the distribution of Lp- PLA2 between one or more lipoproteins may be used to guide treatment, indicating the need or benefit of drug or alternative therapies in the prevention and/or treatment of disorders including atherosclerosis, acute and/or chronic inflammation, cardiac disease, coronary heart disease and stroke.
  • Patients that may benefit from such testing and/or monitoring may include those displaying one or more of: moderate risk of cardiovascular disease (>2 risk factors and Framingham 10-year risk score ⁇ 20%), high risk of cardiovascular disease
  • coronary artery disease or risk equivalents e.g., symptomatic carotid artery disease, peripheral artery disease, abdominal aortic aneurysm, diabetes mellitus
  • moderate risk of stroke e.g., symptomatic carotid artery disease, peripheral artery disease, abdominal aortic aneurysm, diabetes mellitus
  • detection/quantification methods may be used. For example, an enriched sample of Lp-PLA2 may be collected (e.g., by immunoprecipitation as described herein), followed by liquid chromatgrophy/mass spectroscopy (LC-MS), western blotting, or flow cytometry, which may allow both identification of associated lipoproteins as well as quantification. Such methods may also identify non-lipoprotein (including any other proteins/metabolite) that interacts with Lp- PLA2 and may contribute to its clinical/pharmacological/diagnostic utility. For example:
  • immunoaffinity of Lp-PLA2 as described herein may be followed by mass spectroscopy (MS) analysis to identify associated lipids/metabolites/proteins/peptides that interact in a regulated manner under a variety of health/disease/treatment conditions.
  • MS mass spectroscopy
  • Other examples of secondary detection methods that may be used on the enriched Lp-PLA2 samples described herein include ELISAs (direct and competitive, as mentioned above), TIA, Lateral-flow
  • first and second may be used herein to describe various features/elements, these features/elements should not be limited by these terms, unless the context indicates otherwise. These terms may be used to distinguish one feature/element from another feature/element. Thus, a first feature/element discussed below could be termed a second feature/element, and similarly, a second feature/element discussed below could be termed a first feature/element without departing from the teachings of the present invention.
  • numeric value may have a value that is +/- 0.1 % of the stated value (or range of values), +/- 1% of the stated value (or range of values), +/- 2% of the stated value (or range of values), +/- 5% of the stated value (or range of values), +/- 10% of the stated value (or range of values), etc. Any numerical range recited herein is intended to include all sub-ranges subsumed therein.

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Abstract

L'invention concerne des dosages (et en particulier des dosages immunologiques) qui pourvoient à un procédé sensible et fortement reproductible visant à déterminer des niveaux de Lp-PLA2 spécifiques aux lipoprotéines dans des échantillons de patients et/ou les niveaux relatifs de Lp-PLA2 associés à différentes lipoprotéines. L'invention concerne les configurations de dosages dans lesquelles une molécule de capture reconnaissant Lp-PLA2, qui peut être liée à un substrat, et une molécule ou un ensemble de molécules de détection individuelle reconnaissant une sous-espèce de lipoprotéine (par exemple, Apo-Al, Apo-B et apo[a]) sont utilisées pour déterminer la relation entre Lp-PLA2 et une ou plusieurs lipoprotéines.
PCT/US2014/061259 2013-10-18 2014-10-17 Procédé pour la détection d'une association lp-pla2 spécifique aux lipoprotéines Ceased WO2015058158A1 (fr)

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CN106771147A (zh) * 2016-12-22 2017-05-31 广州华弘生物科技有限公司 一种快速诊断脂蛋白相关磷脂酶a2试剂盒及其使用方法
CN106771147B (zh) * 2016-12-22 2018-06-05 广州华弘生物科技有限公司 一种快速诊断脂蛋白相关磷脂酶a2试剂盒及其使用方法
CN111662387A (zh) * 2020-07-14 2020-09-15 重庆中元汇吉生物技术有限公司 抗人脂蛋白相关磷脂酶a2单克隆抗体及其应用

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