CN115494232B - Application of 4,4-dimethyl oxazoline in preparation of reagent containing horseradish peroxidase - Google Patents
Application of 4,4-dimethyl oxazoline in preparation of reagent containing horseradish peroxidase Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title description 8
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- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 2
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Classifications
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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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
Abstract
The application relates to an application of 4,4-dimethyl oxazoline in preparing a reagent containing horseradish peroxidase. The inventor of the application discovers that the 4,4-dimethyl oxazoline can obviously improve the stability of horseradish peroxidase and horseradish peroxidase labeled antibodies in a solution. Therefore, the application of the 4,4-dimethyl oxazoline in preparing the reagent containing the horseradish peroxidase can improve the stability of the horseradish peroxidase in the reagent containing the horseradish peroxidase and widen the application field of the horseradish peroxidase.
Description
Technical Field
The application relates to the technical field of immunodetection, in particular to application of 4,4-dimethyl oxazoline in preparation of a reagent containing horseradish peroxidase.
Background
Horseradish peroxidase (horseradish peroxidase, HRP) is a secreted plant peroxidase derived from horseradish, and is also the most well-studied enzyme among members of the peroxidase family, which contains a single heme and has a plant peroxidase molecular weight of about 44kDa. Horseradish peroxidase can decompose its natural substrate hydrogen peroxide, and two molecules of hydrogen peroxide into water and oxygen, and this redox reaction is achieved in the presence of an electron transfer intermediate, typically an aromatic ring-containing compound. Horseradish peroxidase can be used as a reagent for tissue synthesis and bioconversion, and is commonly applied to the fields of enzyme analysis, chemiluminescence test, immunoassay, bioremediation and the like.
Horseradish peroxidase has a significant disadvantage as a mature, efficient and widely used marker tracer: the stability of the solution is not high. Horseradish peroxidase is not only stable as small molecules like ruthenium terpyridyl, acridinium esters, etc., but also far from alkaline phosphatase (AP enzyme) which is an important marker. The stability problem of horseradish peroxidase limits its application and also reduces the performance of the corresponding immunoassay kit. In particular, when used as a clinical diagnostic reagent, the decreasing activity of horseradish peroxidase causes the measured result to become lower, which makes it necessary for the user to shorten the interval between calibration, thus causing waste and inconvenience.
Therefore, it is important to study how to improve the stability of horseradish peroxidase and horseradish peroxidase-labeled antibodies. The current methods focus mainly on selecting the proper buffer medium, optimal pH, adding bovine serum albumin, etc. However, these methods are difficult to greatly improve the stability of horseradish peroxidase, and even if various existing methods are adopted, the stability of horseradish peroxidase solution is still not ideal, and the half-life period of the signal is about half a year. Therefore, how to improve the stability of horseradish peroxidase and horseradish peroxidase-labeled antibodies has been a difficult problem in the industry.
Disclosure of Invention
The inventor of the application discovers that the 4,4-dimethyl oxazoline can obviously improve the stability of horseradish peroxidase and horseradish peroxidase labeled antibodies in a solution. Based on the above, an embodiment of the application provides an application of 4,4-dimethyl oxazoline in preparing a reagent containing horseradish peroxidase.
Use of 4, 4-dimethyloxazoline in the preparation of a horseradish peroxidase-containing reagent comprising 4, 4-dimethyloxazoline and horseradish peroxidase.
In one embodiment, the horseradish peroxidase-containing reagent comprises 4, 4-dimethyloxazoline and a horseradish peroxidase-labeled antibody.
In one embodiment, the mass percentage of the 4,4-dimethyl oxazoline in the reagent containing horseradish peroxidase is 0.01-1%.
In one embodiment, the horseradish peroxidase-containing reagent further comprises a buffer.
In one embodiment, the buffer comprises one or more of a phosphate buffer, a MES buffer, and an acetate buffer.
In one embodiment, the buffer solution is phosphate buffer solution, and the concentration of the buffer solution is 0.01mol/L to 0.2mol/L.
In one embodiment, the horseradish peroxidase-containing reagent further comprises one or more of bovine serum albumin, polyethylene glycol, and sodium chloride.
In one embodiment, the concentration of the bovine serum albumin in the horseradish peroxidase-containing reagent is 0.1g/L to 10g/L,
in one embodiment, the polyethylene glycol is polyethylene glycol 2000, and the concentration of the polyethylene glycol in the reagent containing horseradish peroxidase is 0.1g/L to 10g/L.
In one embodiment, the concentration of the sodium chloride in the horseradish peroxidase-containing reagent is no more than 50g/L.
In one embodiment, the use of 4-dimethyloxazoline in the preparation of a horseradish peroxidase-containing reagent further comprising a preservative selected from one or more of Proclin300 and BND.
In one embodiment, the application of the 4-dimethyl oxazoline in preparing a reagent containing horseradish peroxidase comprises 0.01 to 0.5 percent of preservative in the reagent containing horseradish peroxidase in percentage by mass.
Drawings
For a clearer description of the technical solutions of the embodiments of the present application, the drawings that are needed in the description of the embodiments will be briefly introduced below, it being obvious that the drawings in the description below are only some embodiments of the present application, and that other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art, wherein:
FIG. 1 is a graph showing the results of measurement of the activity of horseradish peroxidase in example 1.
Detailed Description
The present application will be described more fully hereinafter in order to facilitate an understanding of the application, which may be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
It is noted that, in this document, the term "and/or" includes any and all combinations of one or more of the associated listed items.
The term "antibody" refers to a protein or polypeptide of an immunoglobulin molecule that specifically binds to a corresponding antigen. Antibodies may be polyclonal or monoclonal, multi-chain or single-chain or intact immunoglobulins, derived from natural sources or recombinant sources. For example, naturally occurring IgG antibodies are tetramers comprising at least two heavy (H) chains and two light (L) chains connected to each other by disulfide bonds. Each heavy chain is composed of a heavy chain variable region (VH) and a heavy chain constant region. The heavy chain constant region is composed of three domains (CH 1, CH2 and CH 3). Each light chain is composed of a light chain variable region (VL) and a light chain constant region. The light chain constant region is composed of one domain (CL). VH and VL regions can be further subdivided into regions of hypervariability, termed Complementarity Determining Regions (CDRs), interspersed with regions that are more conserved, termed Framework Regions (FR). Each VH and VL is composed of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of an antibody may mediate the binding of an immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component of the classical complement system (C1 q). "antibodies" include, but are not limited to, monoclonal antibodies, human antibodies, humanized antibodies, chimeric antibodies, and anti-idiotype (anti-Id) antibodies. Antibodies can be of any isotype/class (e.g., igG, igE, igM, igD, igA and IgY) or subclass (e.g., igG1, igG2, igG3, igG4, igA1, and IgA 2).
The term "antigen" as used herein refers to a substance that specifically binds to an antibody. Unless the context contradicts, the terms "antigen" and "test substance" are used interchangeably in the context of the present application. In this context, "optionally" means by way of example, and not necessarily in any way.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the description of the application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.
4, 4-dimethyl-oxazoline (4, 4-dimethyl-oxazolidine) is an organic intermediate useful in the preparation of bromodomain inhibitors. The research shows that 4,4-dimethyl oxazoline has better antibacterial effect on a plurality of bacteria and can be used as a preservative, but the inventor of the application unexpectedly discovers that the effect of improving the stability of horseradish peroxidase by 4,4-dimethyl oxazoline is not caused by the antibacterial capability of the horseradish peroxidase, even if the 4,4-dimethyl oxazoline in the horseradish peroxidase solution subjected to strict sterilization can also exert obvious effect of improving the stability of HRP, and is irrelevant to whether sterilization is carried out, so that the key sites possibly in the horseradish peroxidase structure are combined with the 4,4-dimethyl oxazoline, thereby stabilizing the conformation of protein and improving the stability of horseradish peroxidase. Therefore, the 4,4-dimethyl oxazoline not only can realize the anti-corrosion effect, but also can obviously improve the stability of horseradish peroxidase and horseradish peroxidase labeled antibodies in the solution.
Based on the above, an embodiment of the application provides an application of 4,4-dimethyl oxazoline in preparing a reagent containing horseradish peroxidase.
Specifically, the horseradish peroxidase-containing reagent comprises 4, 4-dimethyloxazoline and horseradish peroxidase. In some embodiments, horseradish peroxidase is present in a separate form (or "free form") in a reagent containing horseradish peroxidase, such as a horseradish peroxidase solution.
In other embodiments, horseradish peroxidase is present in the reagent containing horseradish peroxidase as a label for the antibody, i.e., horseradish peroxidase is labeled on the antibody. The most important application of horseradish peroxidase in the field of in vitro diagnosis is to label antibodies, and the labeled effects such as color development or luminescence are exerted in the final test. If the horseradish peroxidase portion of the labeled antibody gradually fails with the lapse of the storage time, the whole reagent gradually fails. In contrast, the studies of the present application have found that 4, 4-dimethyloxazoline is equally effective on horseradish peroxidase-labeled antibodies. The half life of the reagent can be prolonged from about half a year to about 3 years by adding one thousandth of 4,4-dimethyl oxazoline into the original enzyme-labeled antibody solution system, so that the effective period of the reagent is greatly prolonged.
In some embodiments, the mass percent of 4, 4-dimethyloxazoline in the horseradish peroxidase-containing reagent is 0.01 to 1 percent. In an alternative specific example, the mass percentage of 4, 4-dimethyloxazoline in the horseradish peroxidase-containing reagent is 0.01%, 0.03%, 0.05%, 0.08%, 0.1%, 0.5% or 0.8%. Further, in the reagent containing horseradish peroxidase, the mass percentage of the 4,4-dimethyl oxazoline is 0.01-0.15%. Further, in the reagent containing horseradish peroxidase, the mass percentage of the 4,4-dimethyl oxazoline is 0.07-0.12%.
In some embodiments, the horseradish peroxidase-containing reagent further comprises a buffer. Optionally, the buffer comprises one or more of phosphate buffer, MES buffer, and acetate buffer. It will be appreciated that in other embodiments, the buffer for the horseradish peroxidase-containing reagent is not limited to the above, but may be other buffer systems. Alternatively, the concentration of the buffer solution in the reagent containing horseradish peroxidase is 0.01mol/L to 0.2mol/L. For example, 0.01mol/L, 0.03mol/L, 0.05mol/L, 0.08mol/L, 0.1mol/L, 0.15mol/L or 0.2mol/L.
In one embodiment, the buffer is phosphate buffer, and the concentration of the buffer is 0.01mol/L to 0.2mol/L. Further, the concentration of the phosphoric acid buffer solution is 0.05mol/L to 0.15mol/L. Further, the concentration of the phosphate buffer is 0.08mol/L to 0.12mol/L.
In some embodiments, the horseradish peroxidase-containing reagent of any of the above embodiments further comprises one or more of bovine serum albumin, polyethylene glycol, and sodium chloride.
Alternatively, the concentration of bovine serum albumin in the horseradish peroxidase-containing reagent is 0.1g/L to 10g/L. In an alternative specific example, the concentration of bovine serum albumin in the horseradish peroxidase-containing reagent is 0.1g/L, 0.5g/L, 1g/L, 3g/L, 5g/L, or 8g/L. Further, the concentration of bovine serum albumin in the reagent containing horseradish peroxidase is 1g/L to 5g/L.
Alternatively, the polyethylene glycol is polyethylene glycol 2000. Optionally, the concentration of polyethylene glycol in the horseradish peroxidase-containing reagent is 0.1g/L to 10g/L. In an alternative specific example, the concentration of polyethylene glycol 2000 in the horseradish peroxidase-containing reagent is 0.1g/L, 0.5g/L, 1g/L, 3g/L, 5g/L, or 8g/L. Further, the concentration of polyethylene glycol 2000 in the reagent containing horseradish peroxidase is 0.5g/L to 3g/L.
Alternatively, the concentration of sodium chloride in the horseradish peroxidase-containing reagent is no more than 50g/L. In an alternative specific example, the concentration of sodium chloride in the horseradish peroxidase-containing reagent is 0.5g/L, 1g/L, 10g/L, 20g/L, 30g/L, or 40g/L. Further, the concentration of sodium chloride in the reagent containing horseradish peroxidase is 0.5g/L to 10g/L.
In some embodiments, the horseradish peroxidase-containing reagent further comprises bovine serum albumin, polyethylene glycol 2000 and sodium chloride, wherein the concentration of the bovine serum albumin is 0.1g/L to 10g/L, the concentration of the polyethylene glycol 2000 is 0.1g/L to 10g/L, and the concentration of the sodium chloride is not more than 50g/L. Further, in the reagent containing horseradish peroxidase, the concentration of bovine serum albumin is 1 g/L-5 g/L, the concentration of polyethylene glycol 2000 is 0.5 g/L-3 g/L, and the concentration of sodium chloride is 0.5 g/L-10 g/L.
In some embodiments, the horseradish peroxidase-containing reagent of any of the above embodiments further comprises a preservative. Optionally, the preservative in the horseradish peroxidase-containing reagent is selected from one or more of Proclin300 and 5-bromo-5 nitro-1, 3-dioxane (BND). Optionally, the mass percent of preservative in the reagent containing horseradish peroxidase is 0.01-0.5%. In an alternative specific example, optionally, the preservative is 0.05%, 0.2%, or 0.4% by mass of the horseradish peroxidase-containing reagent. Further, optionally, the preservative in the reagent containing horseradish peroxidase is 0.01-0.4% by mass.
In some embodiments, the pH of the horseradish peroxidase-containing reagent is between 6.5 and 7.5. Further, the pH of the reagent containing horseradish peroxidase is 6.8-7.5.
In some embodiments, the reagent containing horseradish peroxidase-labeled antibody comprises horseradish peroxidase-labeled antibody, 0.01-1% by mass of 4,4-dimethyl oxazoline, 0.01-0.2 mol/L of PBS buffer solution, 0.1-10 g/L of bovine serum albumin, 0.1-10 g/L of polyethylene glycol 2000, not more than 50g/L of sodium chloride, 0.01-0.5% by mass of Proclin300, and pH of 6.5-7.5. Further, the reagent containing the horseradish peroxidase-labeled antibody comprises the horseradish peroxidase-labeled antibody, 0.01-0.15% of 4,4-dimethyl oxazoline, 0.05-0.15 mol/L of PBS buffer solution, 1-5 g/L of bovine serum albumin, 0.5-3 g/L of polyethylene glycol 2000, 0.5-10 g/L of sodium chloride and 0.1-0.5% of Proclin300, wherein the pH is 6.8-7.5.
Based on the above, an embodiment of the application also provides an application of the 4,4-dimethyl oxazoline in preparing a product containing horseradish peroxidase. Specifically, the horseradish peroxidase-containing product comprises the reagent containing the horseradish peroxidase-labeled antibody of any of the above embodiments.
DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION
The following is a detailed description of specific embodiments. The following examples are not specifically described but do not include other components than the unavoidable impurities. Reagents and apparatus used in the examples, unless otherwise specified, are all routine choices in the art. The experimental methods without specific conditions noted in the examples were carried out according to conventional conditions, such as those described in the literature, books, or recommended by the manufacturer. In this context, the expression "M" is a shorthand for mol/L.
EXAMPLE 1 preparation of Horseradish peroxidase solution
1. Preparation of horseradish peroxidase dilution solution: to a 0.1M aqueous PBS (pH 7.4) was added 1g BSA (bovine serum albumin), 0.5g polyethylene glycol 2000, 0.1mL Proclin300 and 0.5g NaCl, and the volume was set to 1 liter to obtain a solution for horseradish peroxidase dilution.
2. And (2) diluting the solid horseradish peroxidase with the horseradish peroxidase prepared in the step (1) to prepare a 10ng/mL solution, and adding different amounts of 4,4-dimethyl oxazoline to obtain horseradish peroxidase solutions containing different concentrations of 4,4-dimethyl oxazoline (the mass percentage of the 4,4-dimethyl oxazoline in the prepared horseradish peroxidase solution is 0.01 percent (or one part per million) and 0.1 percent (or one thousandth), and the horseradish peroxidase solution is used for researching the influence of the 4,4-dimethyl oxazoline on the stability of the horseradish peroxidase.
3. The stability of horseradish peroxidase solutions with varying amounts of 4, 4-dimethyloxazoline added was determined: the method comprises the following specific steps: 1 transparent luminous cup is taken, 100 microliter of luminol luminous solution A is added into the transparent luminous cup, 10 microliter of enzyme solution to be detected is added, then 100 microliter of luminol luminous solution B is added, after rapid sucking and mixing, photographing is immediately carried out after mixing, and the sensitization time is 30 seconds (CR-0302 type optical signal reader), so that the chemiluminescence signal is measured. The luminescence signal initiated in the stability assay was taken as 100% activity. The subsequent luminescence signal was compared with the initial luminescence signal to obtain an activity ratio, and the results are shown in fig. 1. In FIG. 1, the abscissa indicates time (in "day") and the ordinate indicates horseradish peroxidase activity (initial enzyme activity of horseradish peroxidase solution without 4, 4-dimethyloxazoline added was 1, and all horseradish peroxidase solutions were stored under the same conditions at 4 ℃ C.).
As can be seen from FIG. 1, the activity of horseradish peroxidase in a horseradish peroxidase solution containing 0.01% by mass and 0.1% by mass of 4, 4-dimethyloxazoline was maintained at 80% or more or 90% or more after the horseradish peroxidase solution was left for 180 days. However, horseradish peroxidase in horseradish peroxidase solution without addition of 4, 4-dimethyloxazoline had an activity of less than 50% after 180 days of standing. Therefore, the addition of the 4,4-dimethyl oxazoline in the horseradish peroxidase solution can obviously improve the stability of horseradish peroxidase, and the 4,4-dimethyl oxazoline can maintain the stability of the activity of horseradish peroxidase in the horseradish peroxidase solution.
Comparative example 1
The horseradish peroxidase solution in this comparative example was prepared in substantially the same manner as in example 1, except that in this comparative example, 4-dimethyloxazoline was replaced with Proclin300 in equal amounts (0.01% by mass of 4, 4-dimethyloxazoline was replaced with 0.01% by mass of Proclin300, and 0.1% by mass of 4, 4-dimethyloxazoline was replaced with 0.1% by mass of Proclin 300).
The activity of horseradish peroxidase in the horseradish peroxidase solution in this comparative example was measured in the same manner as in example 1.
From the measurement results, it was found that the horseradish peroxidase activities of the group to which 0.01% of Proclin300 was added and the group to which 0.1% of Proclin300 was added were similarly prolonged over time, the enzyme activities were rapidly decreased, and the group to which Proclin300 was additionally added was not significantly different from the control group, as compared with the control group to which Proclin300 was not additionally added.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples merely illustrate a few embodiments of the present application, which are convenient for a specific and detailed understanding of the technical solutions of the present application, but should not be construed as limiting the scope of the claims. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the application, which are all within the scope of the application. It should be understood that, based on the technical solutions provided by the present application, those skilled in the art can obtain technical solutions through logical analysis, reasoning or limited experiments, which are all within the scope of protection of the appended claims. The scope of the patent is therefore intended to be covered by the appended claims, and the description and drawings may be interpreted as illustrative of the contents of the claims.
Claims (12)
- The application of 1.4,4-dimethyl oxazoline in improving the stability of horseradish peroxidase is characterized in that 4,4-dimethyl oxazoline is used for preparing a reagent containing horseradish peroxidase, and the reagent containing horseradish peroxidase comprises 4,4-dimethyl oxazoline and horseradish peroxidase;wherein the stability of the 4, 4-dimethyloxazoline to promote horseradish peroxidase is not caused by the bacteriostatic ability of the 4, 4-dimethyloxazoline.
- 2. The use according to claim 1, wherein the horseradish peroxidase is labelled with an antibody.
- 3. The use according to claim 1, wherein the mass percentage of 4,4-dimethyl oxazoline in the horseradish peroxidase-containing reagent is 0.01% -1%.
- 4. The use according to any one of claims 1 to 3, wherein the horseradish peroxidase-containing reagent further comprises a buffer.
- 5. The use of claim 4, wherein the buffer comprises one or more of phosphate buffer, MES buffer, and acetate buffer.
- 6. The use according to claim 5, wherein the buffer is phosphate buffer, and the concentration of the buffer is 0.01mol/L to 0.2mol/L.
- 7. The use according to any one of claims 1-3 and 5-6, wherein the horseradish peroxidase-containing reagent further comprises one or more of bovine serum albumin, polyethylene glycol and sodium chloride.
- 8. The use according to claim 7, wherein the horseradish peroxidase-containing reagent further comprises bovine serum albumin, and the concentration of bovine serum albumin in the horseradish peroxidase-containing reagent is 0.1g/L to 10g/L.
- 9. The use according to claim 7, wherein the horseradish peroxidase-containing reagent further comprises polyethylene glycol, the concentration of the polyethylene glycol in the horseradish peroxidase-containing reagent is 0.1g/L to 10g/L, and the polyethylene glycol is polyethylene glycol 2000.
- 10. The use according to claim 7, wherein the horseradish peroxidase-containing reagent further comprises sodium chloride, and wherein the concentration of sodium chloride in the horseradish peroxidase-containing reagent is no more than 50g/L.
- 11. The use according to claim 8, wherein the horseradish peroxidase-containing reagent further comprises a preservative selected from one or both of Proclin300 and BND.
- 12. The use according to claim 11, wherein the preservative in the horseradish peroxidase-containing reagent is 0.01% -0.5% by mass.
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| US6133229A (en) * | 1993-10-08 | 2000-10-17 | The University Of Leeds Innovations, Ltd. | Stabilization of proteins in solution |
| CN106478767A (en) * | 2016-11-04 | 2017-03-08 | 郑州伊美诺生物技术有限公司 | A kind of preparation method of enzyme-labelled antigen antibody |
| CN112285342A (en) * | 2020-09-28 | 2021-01-29 | 北京森康维特生物技术研究所 | Dilution stability protective agent for horseradish peroxidase labeled antibody, preparation method and kit thereof |
| CN112924665A (en) * | 2021-02-19 | 2021-06-08 | 山东莱博生物科技有限公司 | Antibody horseradish peroxidase marker and preparation and application thereof |
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| JP4729254B2 (en) * | 2001-09-10 | 2011-07-20 | メソ スケイル テクノロジーズ，エルエルシー | Assay buffer, composition containing it, and method of use thereof |
| KR101789356B1 (en) * | 2010-10-25 | 2017-10-23 | 에프. 호프만-라 로슈 아게 | Use of signal enhancing compounds in electrochemiluminescence detection |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6133229A (en) * | 1993-10-08 | 2000-10-17 | The University Of Leeds Innovations, Ltd. | Stabilization of proteins in solution |
| CN106478767A (en) * | 2016-11-04 | 2017-03-08 | 郑州伊美诺生物技术有限公司 | A kind of preparation method of enzyme-labelled antigen antibody |
| CN112285342A (en) * | 2020-09-28 | 2021-01-29 | 北京森康维特生物技术研究所 | Dilution stability protective agent for horseradish peroxidase labeled antibody, preparation method and kit thereof |
| CN112924665A (en) * | 2021-02-19 | 2021-06-08 | 山东莱博生物科技有限公司 | Antibody horseradish peroxidase marker and preparation and application thereof |
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