CN110585754A - Reagent mixture for drying, preparation of reagent mixture and drying method - Google Patents
Reagent mixture for drying, preparation of reagent mixture and drying method Download PDFInfo
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- CN110585754A CN110585754A CN201910836129.9A CN201910836129A CN110585754A CN 110585754 A CN110585754 A CN 110585754A CN 201910836129 A CN201910836129 A CN 201910836129A CN 110585754 A CN110585754 A CN 110585754A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D19/00—Degasification of liquids
- B01D19/0042—Degasification of liquids modifying the liquid flow
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D19/00—Degasification of liquids
- B01D19/0042—Degasification of liquids modifying the liquid flow
- B01D19/0052—Degasification of liquids modifying the liquid flow in rotating vessels, vessels containing movable parts or in which centrifugal movement is caused
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D19/00—Degasification of liquids
- B01D19/0073—Degasification of liquids by a method not covered by groups B01D19/0005 - B01D19/0042
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D7/00—Sublimation
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
- B01F23/80—After-treatment of the mixture
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
- B01F23/80—After-treatment of the mixture
- B01F23/804—Drying the mixture
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B05—SPRAYING OR ATOMISING IN GENERAL; APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
- B05D—PROCESSES FOR APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
- B05D1/00—Processes for applying liquids or other fluent materials
- B05D1/26—Processes for applying liquids or other fluent materials performed by applying the liquid or other fluent material from an outlet device in contact with, or almost in contact with, the surface
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B05—SPRAYING OR ATOMISING IN GENERAL; APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
- B05D—PROCESSES FOR APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
- B05D7/00—Processes, other than flocking, specially adapted for applying liquids or other fluent materials to particular surfaces or for applying particular liquids or other fluent materials
- B05D7/24—Processes, other than flocking, specially adapted for applying liquids or other fluent materials to particular surfaces or for applying particular liquids or other fluent materials for applying particular liquids or other fluent materials
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B05—SPRAYING OR ATOMISING IN GENERAL; APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
- B05D—PROCESSES FOR APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
- B05D7/00—Processes, other than flocking, specially adapted for applying liquids or other fluent materials to particular surfaces or for applying particular liquids or other fluent materials
- B05D7/50—Multilayers
- B05D7/52—Two layers
- B05D7/53—Base coat plus clear coat type
- B05D7/534—Base coat plus clear coat type the first layer being let to dry at least partially before applying the second layer
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B05—SPRAYING OR ATOMISING IN GENERAL; APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
- B05D—PROCESSES FOR APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
- B05D7/00—Processes, other than flocking, specially adapted for applying liquids or other fluent materials to particular surfaces or for applying particular liquids or other fluent materials
- B05D7/50—Multilayers
- B05D7/52—Two layers
- B05D7/53—Base coat plus clear coat type
- B05D7/536—Base coat plus clear coat type each layer being cured, at least partially, separately
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B05—SPRAYING OR ATOMISING IN GENERAL; APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
- B05D—PROCESSES FOR APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
- B05D7/00—Processes, other than flocking, specially adapted for applying liquids or other fluent materials to particular surfaces or for applying particular liquids or other fluent materials
- B05D7/50—Multilayers
- B05D7/56—Three layers or more
- B05D7/58—No clear coat specified
- B05D7/584—No clear coat specified at least some layers being let to dry, at least partially, before applying the next layer
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B05—SPRAYING OR ATOMISING IN GENERAL; APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
- B05D—PROCESSES FOR APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
- B05D7/00—Processes, other than flocking, specially adapted for applying liquids or other fluent materials to particular surfaces or for applying particular liquids or other fluent materials
- B05D7/50—Multilayers
- B05D7/56—Three layers or more
- B05D7/58—No clear coat specified
- B05D7/586—No clear coat specified each layer being cured, at least partially, separately
Abstract
The invention discloses a reagent mixture for drying, a preparation method of the reagent mixture and a drying method, wherein the reagent mixture consists of a reagent to be dried and a shaping agent; the volume of the reagent mixture consisting of the reagent to be dried and the shaping agent is V, the mass of the shaping agent is M1, the mass concentration of the shaping agent in the mixture is C1-M1/V, and the value range of C1 is 0.01g/L-50 g/L; according to the invention, the shaping agent is added into the reagent to be dried, so that the appearance of the dried reagent can be optimized, the viscosity of the reagent to be dried can be increased, the reagent to be dried can be conveniently fixed on a platform made of a hydrophobic material or a platform coated with the hydrophobic material or other platforms through the viscous force, and the probability that the dried reagent is powdery due to the fact that the reagent to be dried cannot be completely adhered to the platform, bubbles are generated when the reagent is redissolved, and the bubbles are increased in the heating process, take away components in the reagent to be dried, and influence the detection result of the reagent to be dried after redissolved is reduced.
Description
All as the field of technology
The invention relates to the technical field of reagent drying, in particular to a reagent mixture for drying and a reagent drying method.
All the above-mentioned background techniques
The reagent is dried and stored, which is a common method for storing the reagent in the prior art, and compared with the reagent in a liquid state, the reagent is dried and stored, which has the advantages of long storage time, small occupied space, convenient package, convenient transportation and the like. However, as shown in fig. 1, due to the low viscosity between the reagent and the hydrophobic material, bubbles are generated in the process of drying the reagent on the hydrophobic material and then adding the liquid for redissolution, and the bubbles are increased in the heating process, so that a large amount of bubbles can take away active ingredients in the reagent, thereby affecting the subsequent detection result of the reagent.
All the contents of the invention
The first purpose of the invention is to provide a reagent mixture which is convenient to dry and reduces air bubbles generated when the reagent is redissolved after being dried.
In order to realize the first purpose of the invention, the invention adopts the following technical scheme:
a reagent mixture for drying, consisting of a reagent to be dried and a shaping agent; the volume of the reagent mixture consisting of the reagent to be dried and the shaping agent is V, the mass of the shaping agent is M1, the mass concentration of the shaping agent in the mixture is C1-M1/V, and the value range of C1 is 0.01g/L-50 g/L.
As a specific embodiment, the shaping agent employs at least one of mannitol, gum arabic, pectin, gelatin, or maltodextrin.
The second purpose of the invention is to provide a reagent mixture which is convenient to dry, prolongs the storage life of the reagent mixture and reduces bubbles generated when the reagent is redissolved after being dried.
In order to achieve the second purpose of the invention, the invention adopts the following technical scheme:
a reagent mixture for drying is composed of a reagent to be dried, a shaping agent and a stabilizing agent; the volume of the reagent mixture consisting of the reagent to be dried, the shaping agent and the stabilizing agent is V, the mass of the shaping agent is M1, the mass of the stabilizing agent is M2, the mass concentration of the shaping agent in the mixture is C1-M1/V, and the value range of C1 is 0.01g/L-50 g/L; the mass concentration of the stabilizer in the mixture is C2-M2/V, and the value range of C2 is 0.01g/L-50 g/L.
In a specific embodiment, the stabilizer is composed of an organic substance, preferably at least one of a saccharide, a protein or another stabilizer.
In a specific embodiment, the saccharide is at least one of glucose, trehalose, sucrose, galactose or maltose; the protein is at least one of whey protein isolate, soy protein isolate or bovine serum albumin; the other stabilizer is at least one of glycerol, skimmed milk powder, PVP, Tween and glycine.
Further, the reagent to be dried is composed of at least one of an inorganic substance, an organic substance, an enzyme, and an antibody.
In a specific embodiment, the reagent to be dried is a DNA detection reagent or a protein detection reagent or an immunofluorescence reaction system reagent or a chemiluminescence reaction system reagent.
The third purpose of the invention is to provide a preparation method of the reagent mixture, which can reduce the bubbles generated when the reagent is redissolved after being dried.
In order to achieve the third object of the present invention, the present invention adopts the following technical solutions:
a method of preparing a reagent mixture comprising the steps of: s11, adding the plasticizer or the stabilizer into the reagent to be dried according to the requirement of mass concentration, and stirring; s12, performing at least one of ultrasonic treatment, centrifugal treatment or vacuum treatment on the mixture obtained in the step 11.
The fourth purpose of the invention is to provide a preparation method of the reagent mixture, which can reduce the bubbles generated when the reagent is redissolved after being dried.
In order to achieve the fourth object of the present invention, the present invention adopts the following technical solutions:
a method of drying a reagent mixture comprising the steps of: s21, spotting the reagent mixture on a platform coated with a hydrophobic material or made of a hydrophobic material, S22, drying the spotted mixture on the platform,
or, comprising the steps of: s31, spotting a glue layer on the platform, drying the glue layer, S32, spotting the reagent mixture on the glue layer obtained in the previous step, drying the reagent mixture,
or, comprising the steps of: s41, spotting a glue layer on the platform, and drying the glue layer; s42, spotting the reagent mixture on the glue layer obtained in the previous step and drying the reagent mixture, S43, spotting a glue layer on the reagent mixture obtained in the previous step and drying the glue layer, S44, spotting the reagent mixture on the glue layer obtained in the previous step and drying the reagent mixture,
or, comprising the steps of: s41, spotting a glue layer on the platform, and drying the glue layer; s42, spotting the reagent mixture on the glue layer obtained in the last step and drying the reagent mixture, S43, spotting a glue layer on the reagent mixture obtained in the last step and drying the glue layer, S44, spotting the reagent mixture on the glue layer obtained in the last step and drying the reagent mixture, and repeating the steps S43 and S44 one or more times;
the glue layer is prepared from a sticky material.
A method of drying a reagent mixture comprising the steps of: s31, spotting a glue layer on the platform, drying the glue layer, S32, spotting the reagent mixture on the glue layer obtained in the previous step, drying the reagent mixture,
or, comprising the steps of: s41, spotting a glue layer on the platform, and drying the glue layer; s42, spotting the reagent mixture on the glue layer obtained in the previous step and drying the reagent mixture, S43, spotting a glue layer on the reagent mixture obtained in the previous step and drying the glue layer, S44, spotting the reagent mixture on the glue layer obtained in the previous step and drying the reagent mixture,
or, comprising the steps of: s41, spotting a glue layer on the platform, and drying the glue layer; s42, spotting the reagent mixture on the glue layer obtained in the last step and drying the reagent mixture, S43, spotting a glue layer on the reagent mixture obtained in the last step and drying the glue layer, S44, spotting the reagent mixture on the glue layer obtained in the last step and drying the reagent mixture, and repeating the steps S43 and S44 one or more times;
the reagent mixture consists of a reagent to be dried and a shaping agent; the adhesive layer consists of a shaping agent; the volume of each sample application of the reagent mixture is V0, wherein the mass concentration of the plasticizer is C1, and the mass of the plasticizer used for each sample application of the glue layer is M0, (C1V 0+ M0)/V0 is in the range of 0.01g/L-50 g/L.
A method of drying a reagent mixture comprising the steps of: s31, spotting a glue layer on the platform, drying the glue layer, S32, spotting the reagent mixture on the glue layer obtained in the previous step, drying the reagent mixture,
or, comprising the steps of: s41, spotting a glue layer on the platform, and drying the glue layer; s42, spotting the reagent mixture on the glue layer obtained in the last step and drying the reagent mixture, S43, spotting a glue layer on the reagent mixture obtained in the last step and drying the glue layer, S44, spotting the reagent mixture on the glue layer obtained in the last step and drying the reagent mixture, and repeating the steps S43 and S44 one or more times;
the reagent mixture consists of a reagent to be dried, a shaping agent and a stabilizing agent; the adhesive layer consists of a shaping agent; the volume of the reagent mixture is V1, wherein the mass concentration of the plasticizer is C1, and the mass concentration of the stabilizer is C2; the volume of each sample application of the reagent mixture is V0, the mass M0 of the shaping agent used for each sample application of the glue layer is equal to C1V 0, the number of the glue layers is N, the value range of (C1V 0+ N M0)/V0 is 0.01g/L-50g/L, and the value range of C2 is 0.01g/L-50 g/L.
Further, the number of spotting layers of the reagent mixture ranges from 1 to 50 layers, preferably from 1 to 5 layers.
As a specific embodiment, the drying treatment is at least one of freeze drying, drying or vacuum drying.
The invention has the beneficial effects that:
according to the invention, the shaping agent is added into the reagent to be dried, so that the appearance of the dried reagent can be optimized, the viscosity of the reagent to be dried can be increased, the reagent to be dried can be conveniently fixed on a platform made of a hydrophobic material or a platform coated with the hydrophobic material or a platform made of other materials through viscosity force, the dried reagent is powdery or separated from the platform to generate gaps due to the fact that the reagent to be dried cannot be completely adhered to the platform, when liquid is added for redissolution, bubbles can be generated in the reagent, and can be increased in the heating process, a large number of bubbles can take away components in the reagent to be dried, and the detection result of the redissolved reagent to be dried is influenced. Further, the stabilizing agent and the shaping agent are added into the reagent to be dried, so that the storage life of a reagent mixture can be prolonged, the shaping agent can optimize the appearance of the dried reagent and increase the viscosity of the reagent to be dried, the reagent to be dried is conveniently fixed on a platform made of a hydrophobic material or a platform coated with the hydrophobic material through the viscous force, the problem that the dried reagent is powdery or separated from the platform to generate gaps due to the fact that the reagent to be dried cannot be completely adhered to the platform is reduced, bubbles can be generated when liquid is added for heating and redissolving, the bubbles can be increased in the heating process, a large number of bubbles can take away components in the reagent to be dried, and the detection result after the reagent to be dried is redissolved is influenced; furthermore, the invention achieves the purpose of removing air in the reagent mixture by carrying out ultrasonic treatment, centrifugal treatment or vacuum pumping treatment on the uniformly mixed reagent before the reagent is dried, and reduces the probability of generating bubbles after the dried reagent is redissolved; furthermore, the reagent mixture is spotted on the platform for drying, so that the reagent mixture is convenient to transport; furthermore, the dried reagent mixture is adhered to the platform through the adhesive layer, so that bubbles generated after the reagent mixture is redissolved are reduced; further, the invention completes the drying treatment by freeze-drying, drying or vacuumizing.
Description of the drawings
In order to more clearly illustrate the embodiments of the present invention, the drawings used in the embodiments will be briefly described below. The drawings in the following description are only embodiments of the invention and other drawings may be derived from those drawings by a person skilled in the art without inventive effort.
FIG. 1 is a graph of the reconstitution results of a prior art reagent of the present invention after drying;
FIG. 2 is a flow chart of a method of preparing a reagent mixture according to an embodiment of the present invention;
FIG. 3 is a flow chart of a method for drying a reagent mixture according to an embodiment of the present invention;
FIG. 4 is a graph showing the reconstitution results of a dried reagent according to one embodiment of the present invention;
FIG. 5 is a flow chart of a method for drying a reagent mixture according to example two of the present invention;
FIG. 6 is a graph showing the results of reconstitution after drying of reagents according to example two of the present invention;
FIG. 7 is a graph showing the results of reconstitution after drying of the third reagent in the example of the present invention;
FIG. 8 is a flow chart of a method for drying a reagent mixture according to an embodiment of the present invention;
FIG. 9 is a graph showing the results of reconstitution of a dried reagent of the fourth embodiment of the present invention;
FIG. 10 is a graph showing the results of reconstitution after drying of five reagents according to example of the present invention;
FIG. 11 is a graph showing the results of reconstitution after drying of six reagents according to examples of the present invention.
(specific embodiments) in all cases
The present invention will be described in detail below with reference to the accompanying drawings.
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example one
A reagent mixture for drying is composed of a reagent to be dried, a shaping agent and a stabilizing agent; the total volume of the reagent mixture is V, wherein the mass of the shaping agent is M1, the mass of the stabilizing agent is M2, the mass concentration of the shaping agent in the reagent mixture is C1, C1 is M1/V, the mass concentration of the stabilizing agent in the reagent mixture is C2, and C2 is M2/V.
In this example, the reagent to be dried is a DNA detection reagent prepared from HBVPCR reaction solution, Taq enzyme and UDG enzyme in a hepatitis b virus nucleic acid amplification (PCR) fluorescence quantitative detection kit (20 persons/cassette) manufactured by bosch technologies ltd, hangzhou, and the volume ratio of the HBV PCR reaction solution, the Taq enzyme and the UDG enzyme is 377: 3: 1. In this example, the volume of HBV PCR reaction solution used for preparing DNA detection reagent was 37.7ul, the volume of Taq enzyme was 0.3ul, and the volume of UDG enzyme was 0.1 ul. When preparing the reagent mixed solution, taking out the reagent to be dried with the required volume from the prepared reagent to be dried, and adding the stabilizer and the shaping agent into the taken-out reagent to be dried according to the requirements of the mass concentration of the stabilizer and the shaping agent.
In the embodiment, galactose is adopted as the stabilizing agent, and the mass concentration of the stabilizing agent in the reagent mixture is 0.01 g/L; in other embodiments, the stabilizer is at least one of a saccharide, glucose, trehalose, sucrose, galactose or maltose, or a protein such as at least one of whey protein isolate, soy protein isolate or bovine serum albumin, or other stabilizer such as at least one of glycerol, skim milk powder, PVP, tween, glycine.
In this example, gum arabic is used as the shaping agent; in other embodiments, the shaping agent employs at least one of mannitol, acacia, gelatin, or maltodextrin.
In this embodiment, the solute in the HBV PCR reaction solution is inorganic, Taq enzyme and UDG enzyme are enzymes, and the reagent to be dried is composed of inorganic and enzyme; the sugar, protein and other stabilizers in the stabilizer are all organic matters.
As shown in fig. 1, in this embodiment, a method for preparing a reagent mixture includes the following steps: s11, adding the plasticizer or the stabilizer into the reagent to be dried according to the requirement of mass concentration, and stirring; s12, carrying out ultrasonic treatment on the mixture obtained in the step 12; s13, centrifuging the mixture obtained in the step 12; and S14, performing vacuum-pumping treatment on the mixture in the step 13.
In this example, step S12, the frequency of the ultrasonic treatment of the reagent mixture in the ultrasonic machine is 30KHZ, and the time is 5 min: step S12, placing the reagent mixture into a centrifuge for centrifugation at 3500-4500rpm for 5 min; in step S12, the reagent mixture is placed in a vacuum machine for vacuuming for 5 min.
In other embodiments, one or both of sonication, centrifugation, and evacuation of the uniformly mixed reagent mixture; ultrasonic treatment, centrifugal treatment and vacuum pumping treatment can all achieve the purpose of removing air in the reagent mixture, and reduce the probability of generating bubbles after redissolving the dried reagent.
In the embodiment, the shaping agent is added into the reagent to be dried, so that the appearance of the dried reagent can be optimized, the viscosity of the reagent to be dried can be increased, the reagent to be dried can be conveniently fixed on a platform made of a hydrophobic material or a platform coated with the hydrophobic material through viscous force, the dried reagent is powdery due to the fact that the reagent to be dried cannot be completely adhered to the platform, bubbles can be generated when liquid is added for redissolving, components in the reagent to be dried are taken away, and the detection result after the reagent to be dried is redissolved is influenced; the shelf life of the reagent mixture can be extended by adding a stabilizer to the reagent to be dried.
In this example, the shelf life of the reagent mixture may be up to 12 months.
As shown in fig. 2, in this embodiment, a method for drying a reagent mixture includes the following steps: s31, spotting a glue layer on the platform, and drying the glue layer, S32, spotting the reagent mixture on the glue layer obtained in the previous step, and drying the reagent mixture.
In this embodiment, the glue layer is composed of a molding agent; the volume V0 of each spot of the reagent mixture was equal to 2ul, and the mass M10 of the shaping agent was equal to 2 × C1 for each spot of the reagent mixture; when the glue layer is spotted, the mass of the plasticizer in the glue layer is M0, and the ratio of (M0+ M10)/V0 is 0.01 g/L.
In this example, the mass concentration C1 of the plasticizer in the reagent mixture is equal to 0.005g/L, and the mass M0 of the plasticizer spotted on the gel layer is equal to 0.01 ug. In other embodiments, the mass concentration C1 of the plasticizer in the reagent mixture is equal to 0.004g/L and the mass M0 of the gel-spotted plasticizer is equal to 0.012 ug.
In this embodiment, a glue layer is disposed between the dried reagent mixture and the platform; the two surfaces of the adhesive layer, which are respectively contacted with the reagent mixture and the platform, are provided with viscosity for fixing and adhering the reagent mixture on the platform.
As shown in fig. 3, compared with the prior art, in the present embodiment, the air bubbles generated after the redissolution of the reagent mixture are reduced by adding the plasticizer to the reagent to be dried and adhering the dried reagent mixture to the platform through the adhesive layer.
In this embodiment, the platform is a substrate coated with a hydrophobic layer in the microfluidic chip.
In this embodiment, the drying process adopts freeze-drying, and includes the following steps: pre-freezing, namely pre-freezing the platform subjected to sample application (sample application of a reagent mixture or sample application of a glue layer) in a refrigerator at the temperature of-20 ℃ for 8 hours; placing the pre-frozen platform on a shelf of a freeze dryer; carrying out first sublimation, vacuumizing the freeze dryer to 10pa, setting the temperature of a shelf at-20 ℃, and keeping for 6 hours; for the second sublimation, the freeze dryer was evacuated to 1pa and the shelf temperature was raised to 20 ℃ for 4 hours.
Example two
The difference between this embodiment and the first embodiment is: the reagent to be dried is a protein detection reagent, and is prepared by adopting a BCA reagent A and a BCA reagent B in a BCA protein quantitative kit (PA115-01) produced by Tiangen Biochemical technology (Beijing) Limited company according to the volume ratio of 50: 1; as shown in fig. 5, the drying method of the reagent mixture includes the following steps: s41, spotting a glue layer on the platform, and drying the glue layer; s42, spotting the reagent mixture on the glue layer obtained in the last step, and drying the reagent mixture, S43, spotting a glue layer on the reagent mixture obtained in the last step, and drying the glue layer, S44, spotting the reagent mixture on the glue layer obtained in the last step, and drying the reagent mixture, repeating the steps S43 and S44, and performing the steps S43 and S44 for four times, namely, each spotting point is finally overlapped with 5 layers of dried reagent mixtures through 5 layers of glue layers; the mass concentration of the stabilizer in the reagent mixture is 10 g/L; the ratio of the sum of the mass of the plasticizer in the 5 layers of glue and 5 layers of reagent mixtures to the volume of the five spotted reagent mixtures, i.e. (M0 × 5+ M10 × 5)/(5 × V0) ═ 10g/L, i.e., (M0+ M10)/(V0) ═ 10 g/L.
In this example, the mass concentration C1 of the plasticizer in the reagent mixture is equal to 5g/L, and the mass M0 of the plasticizer per one spotting of the glue layer is equal to 10 ug. In other embodiments, the mass concentration of plasticizer C1 in the reagent mixture is equal to 4g/L and the mass of the gel-spotted plasticizer M0 is equal to 12 ug.
In this example, the reagents to be dried were formulated with a volume of BCA reagent a of 50ul and a volume of BCA reagent B of 1 ul. When preparing the reagent mixed solution, taking out the reagent to be dried with the required volume from the prepared reagent to be dried, and then adding the stabilizer and the plasticizer into the reagent according to the requirement of the mass concentration of the stabilizer and the plasticizer.
In this example, the BCA reagent a and the BCA reagent B contain Bovine Serum Albumin (BSA) solution as a protein standard solution, Bovine Serum Albumin (BSA) is an organic substance, and the reagents to be dried are composed of an organic substance.
As shown in fig. 6, in this embodiment, compared with the prior art, the dried reagent mixture is spotted in multiple layers through glue layers, and the dried reagent mixture is adhered to the platform, so that bubbles generated after the reagent mixture is redissolved are reduced.
In this embodiment, the drying process adopts drying, and includes the following steps: the platform after sample application (reagent mixture sample application or glue layer sample application) is placed in a blast drying oven at 60 ℃ and dried for 60 min.
In the third embodiment, the first step is that,
the difference between this embodiment and the second embodiment is: the reagent to be dried is an immunofluorescence reaction system reagent and is prepared by adopting a buffer solution in a Myoglobin (MYO) detection kit (25 persons/box) produced by Beijing KangXingrun biotechnology limited; the steps S43 and S44 are performed 49 times in total, that is, 50 layers of dried reagent mixture are finally stacked on each sample addition point through 50 layers of glue layers; the mass concentration of the stabilizer in the reagent mixture is 50 g/L; the ratio of the sum of the mass of the shaping agent in 50 layers of glue and 50 layers of reagent mixture to the volume of the reagent mixture spotted at 50 times, i.e. (M0 × 50+ M10 × 50)/(50 × V0) ═ 50g/L, i.e., (M0+ M10)/(V0) ═ 50 g/L.
In this example, the buffer consisted of sodium salt, BSA, sugar, surfactant, fluorescently labeled MYO monoclonal antibody II, rabbit anti-chicken antibody, etc.; the sodium salt is an inorganic substance, BSA, sugar and a surfactant are organic substances, the fluorescence-labeled MYO monoclonal antibody II and chicken antibody are antibodies, and the reagent to be dried is composed of the inorganic substance, the organic substance and the antibodies.
In this example, the mass concentration C1 of the plasticizer in the reagent mixture is equal to 25g/L, and the mass M0 of the plasticizer per one spotting of the glue layer is equal to 50 ug. In other embodiments, the mass concentration of plasticizer C1 in the reagent mixture is equal to 20g/L and the mass of the gel-spotted plasticizer M0 is equal to 60 ug.
In this example, the reagents to be dried were prepared with a buffer volume of 50 ul. When preparing the reagent mixed solution, taking out the reagent to be dried with the required volume from the prepared reagent to be dried, and then adding the stabilizer and the plasticizer into the reagent according to the requirement of the mass concentration of the stabilizer and the plasticizer.
As shown in fig. 7, in this embodiment, compared with the prior art, the dried reagent mixture is spotted in multiple layers through glue layers, and the dried reagent mixture is adhered to the platform, so that bubbles generated after the reagent mixture is redissolved are reduced.
In this embodiment, the drying process adopts vacuum drying, and includes the specific steps of: the platform after sample application (sample application of the reagent mixture or the glue layer) is treated for 10min at 4 ℃ in vacuum.
Example four
The difference between this embodiment and the first embodiment is: the reagent to be dried is a chemiluminescence reaction system reagent, and is prepared by labeling and concentrating a working concentration mark of progesterone (Pro) in a progesterone (Pro) chemiluminescence kit (96 parts) produced by Jinan Xingyen biotechnology limited; as shown in fig. 8, the drying method of the reagent mixture includes the steps of: s21 spotting the reagent mixture on a platform coated with a hydrophobic material or made of a hydrophobic material, S22 drying the spotted mixture on the platform; in the reagent mixture, the mass concentration of the plasticizer is 0.01 g/L.
In this example, the progesterone (Pro) working concentration labeled concentrated conjugate solution contains Anti-Pro (labeled conjugate solution), 1% BSA, PBS (phosphate buffered saline) PH 7.4-7.5; PBS is inorganic matter, Anti-Pro and BSA are organic matter, and the reagent to be dried is composed of inorganic matter and organic matter.
In this example, reagents to be dried were prepared using a volume of 100ul of progesterone (Pro) working concentration labeled concentrated coupling solution. When preparing the reagent mixed solution, taking out the reagent to be dried with the required volume from the prepared reagent to be dried, and adding the stabilizing agent and the plasticizer into the reagent to be dried according to the mass concentration of the stabilizing agent and the plasticizer.
As shown in fig. 9, compared with the prior art, the present embodiment increases the viscosity of the reagent to be dried by adding the plasticizer to the reagent to be dried, and reduces the bubbles after the re-dissolution of the reagent mixture.
EXAMPLE five
The present embodiment is different from the fourth embodiment in that: in the reagent mixture, the mass concentration of the plasticizer is 10 g/L.
As shown in fig. 10, compared with the prior art, the present embodiment increases the viscosity of the reagent to be dried by adding the plasticizer to the reagent to be dried, and reduces the bubbles after the re-dissolution of the reagent mixture.
EXAMPLE six
The present embodiment is different from the fourth embodiment in that: in the reagent mixture, the mass concentration of the plasticizer is 50 g/L.
As shown in fig. 11, compared with the prior art, the present embodiment increases the viscosity of the reagent to be dried by adding the plasticizer to the reagent to be dried, and reduces the bubbles after the re-dissolution of the reagent mixture.
The above embodiments are only preferred embodiments of the present invention, and the protection scope of the present invention is not limited to the above embodiments, and all technical solutions belonging to the idea of the present invention belong to the protection scope of the present invention. It should be noted that modifications and embellishments within the scope of the invention may occur to those skilled in the art without departing from the principle of the invention, and are considered to be within the scope of the invention.
Claims (13)
1. A reagent mixture for drying, characterized by: consists of a reagent to be dried and a shaping agent; the volume of the reagent mixture consisting of the reagent to be dried and the shaping agent is V, the mass of the shaping agent is M1, the mass concentration of the shaping agent in the mixture is C1-M1/V, and the value range of C1 is 0.01g/L-50 g/L.
2. The reagent mixture of claim 1, wherein: the plastic agent is at least one of mannitol, Arabic gum, pectin, gelatin or maltodextrin.
3. A reagent mixture for drying, characterized by: consists of a reagent to be dried, a shaping agent and a stabilizing agent; the volume of the reagent mixture consisting of the reagent to be dried, the shaping agent and the stabilizing agent is V, the mass of the shaping agent is M1, the mass of the stabilizing agent is M2, the mass concentration of the shaping agent in the mixture is C1-M1/V, and the value range of C1 is 0.01g/L-50 g/L; the mass concentration of the stabilizer in the mixture is C2-M2/V, and the value range of C2 is 0.01g/L-50 g/L.
4. The reagent mixture of claim 3, wherein: the stabilizer adopts at least one of saccharide, protein or other stabilizers.
5. The reagent mixture of claim 4, wherein: the saccharide is at least one of glucose, trehalose, sucrose, galactose or maltose; the protein is at least one of whey protein isolate, soy protein isolate or bovine serum albumin; the other stabilizer is at least one of glycerol, skimmed milk powder, PVP, Tween and glycine.
6. The reagent mixture of any one of claims 1-5, wherein: the reagent to be dried is composed of at least one of an inorganic substance, an organic substance, an enzyme, and an antibody.
7. The reagent mixture of claim 6, wherein: the reagent to be dried is a DNA detection reagent or a protein detection reagent or an immunofluorescence reaction system reagent or a chemiluminescence reaction system reagent.
8. A method for preparing a reagent mixture according to any one of claims 1 to 7, comprising the steps of: s11, adding the plasticizer or the stabilizer into the reagent to be dried according to the requirement of mass concentration, and stirring; s12, at least one of ultrasonic treatment, centrifugal treatment or vacuum pumping treatment is carried out on the mixture obtained in the step S11.
9. A method of drying a reagent mixture according to any one of claims 1 to 7, comprising the steps of: s21, spotting the reagent mixture on a platform coated with a hydrophobic material or made of a hydrophobic material, S22, drying the spotted mixture on the platform,
or, comprising the steps of: s31, spotting a glue layer on the platform, drying the glue layer, S32, spotting the reagent mixture on the glue layer obtained in the previous step, drying the reagent mixture,
or, comprising the steps of: s41, spotting a glue layer on the platform, and drying the glue layer; s42, spotting the reagent mixture on the glue layer obtained in the previous step and drying the reagent mixture, S43, spotting a glue layer on the reagent mixture obtained in the previous step and drying the glue layer, S44, spotting the reagent mixture on the glue layer obtained in the previous step and drying the reagent mixture,
or, comprising the steps of: s41, spotting a glue layer on the platform, and drying the glue layer; s42, spotting the reagent mixture on the glue layer obtained in the last step and drying the reagent mixture, S43, spotting a glue layer on the reagent mixture obtained in the last step and drying the glue layer, S44, spotting the reagent mixture on the glue layer obtained in the last step and drying the reagent mixture, and repeating the steps S43 and S44 one or more times;
the glue layer is prepared from a sticky material.
10. A method of drying a reagent mixture according to any one of claims 1 to 7, characterized in that: the method comprises the following steps: s31, spotting a glue layer on the platform, drying the glue layer, S32, spotting the reagent mixture on the glue layer obtained in the previous step, drying the reagent mixture,
or, comprising the steps of: s41, spotting a glue layer on the platform, and drying the glue layer; s42, spotting the reagent mixture on the glue layer obtained in the previous step and drying the reagent mixture, S43, spotting a glue layer on the reagent mixture obtained in the previous step and drying the glue layer, S44, spotting the reagent mixture on the glue layer obtained in the previous step and drying the reagent mixture,
or, comprising the steps of: s41, spotting a glue layer on the platform, and drying the glue layer; s42, spotting the reagent mixture on the glue layer obtained in the last step and drying the reagent mixture, S43, spotting a glue layer on the reagent mixture obtained in the last step and drying the glue layer, S44, spotting the reagent mixture on the glue layer obtained in the last step and drying the reagent mixture, and repeating the steps S43 and S44 one or more times;
the reagent mixture consists of a reagent to be dried and a shaping agent; the adhesive layer consists of a shaping agent; the volume of each sample application of the reagent mixture is V0, wherein the mass concentration of the plasticizer is C1, and the mass of the plasticizer used for each sample application of the glue layer is M0, (C1V 0+ M0)/V0 is in the range of 0.01g/L-50 g/L.
11. A method of drying a reagent mixture according to any one of claims 1 to 7, characterized in that: the method comprises the following steps: s31, spotting a glue layer on the platform, drying the glue layer, S32, spotting the reagent mixture on the glue layer obtained in the previous step, drying the reagent mixture,
or, comprising the steps of: s41, spotting a glue layer on the platform, and drying the glue layer; s42, spotting the reagent mixture on the glue layer obtained in the last step and drying the reagent mixture, S43, spotting a glue layer on the reagent mixture obtained in the last step and drying the glue layer, S44, spotting the reagent mixture on the glue layer obtained in the last step and drying the reagent mixture, and repeating the steps S43 and S44 one or more times;
the reagent mixture consists of a reagent to be dried, a shaping agent and a stabilizing agent; the adhesive layer consists of a shaping agent; the volume of the reagent mixture is V1, wherein the mass concentration of the plasticizer is C1, and the mass concentration of the stabilizer is C2; the volume of each sample application of the reagent mixture is V0, the mass M0 of the shaping agent used for each sample application of the glue layer is equal to C1V 0, the number of the glue layers is N, the value range of (C1V 0+ N M0)/V0 is 0.01g/L-50g/L, and the value range of C2 is 0.01g/L-50 g/L.
12. Drying method according to any of claims 9 to 11, wherein the number of spotting layers of the reagent mixture is in the range of 1 to 50 layers, preferably 1 to 5 layers.
13. Drying method according to any one of claims 9 to 11, characterised in that: the drying treatment adopts at least one of freeze drying, drying or vacuum drying.
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Correction item: Applicant|Address|Applicant Correct: Zhuhai diqifurui Biotechnology Co., Ltd|519000No. 2522 Huandao North Road, Hengqin New District, Zhuhai City, Guangdong Province, business service center-115 (centralized office area) of Guangdong Macao cooperative traditional Chinese medicine science and Technology Industrial Park|postdoctoral research station of Guangzhou Nansha Information Technology Park Co., Ltd False: Zhuhai diqifurui Biotechnology Co., Ltd|519000No.2522 Huandao North Road, Hengqin New District, Zhuhai City, Guangdong Province, business service center-115 (centralized office area) of Guangdong Macao cooperative traditional Chinese medicine science and Technology Industrial Park|postdoctoral research station of Guangzhou Nansha information science and Technology Park Co., Ltd Number: 51-02 Volume: 35 Correction item: Applicant|Address|Applicant Correct: Zhuhai diqifurui Biotechnology Co., Ltd|519000 No. 2522 Huandao North Road, Hengqin New District, Zhuhai City, Guangdong Province, business service center-115 (centralized office area) of Guangdong Macao cooperative traditional Chinese medicine science and Technology Industrial Park|postdoctoral research station of Guangzhou Nansha Information Technology Park Co., Ltd False: Zhuhai diqifurui Biotechnology Co., Ltd|519000 No.2522 Huandao North Road, Hengqin New District, Zhuhai City, Guangdong Province, business service center-115 (centralized office area) of Guangdong Macao cooperative traditional Chinese medicine science and Technology Industrial Park|postdoctoral research station of Guangzhou Nansha information science and Technology Park Co., Ltd Number: 51-02 Page: The title page Volume: 35 |
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