CN101363846A - Quality-control product of drying human lymphocyte surface antigen and method for making same - Google Patents
Quality-control product of drying human lymphocyte surface antigen and method for making same Download PDFInfo
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- CN101363846A CN101363846A CNA200810150909XA CN200810150909A CN101363846A CN 101363846 A CN101363846 A CN 101363846A CN A200810150909X A CNA200810150909X A CN A200810150909XA CN 200810150909 A CN200810150909 A CN 200810150909A CN 101363846 A CN101363846 A CN 101363846A
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Abstract
The invention relates to a product for controlling the quality of lyophilized human lymphocyte surface antigens, which is a grey-white solid prepared by lyophilizing a cell suspension having a cell concentration of 400,000-600,000/mL and containing lyophilized human lymphocytes immobilized by a neutral buffer formaldehyde solution and a protective agent. The mark values respectively are: CD<3> of 37.25-69.1%, CD<4> of 17.9-33.7%, CD<8> of 12.4-23.9%, and the ratio of CD<4>/CD<8> of 0.98-1.94. The invention also provides the preparation method of the quality control product. The product has the characteristics of good antigen storability and stable antigenicity, and can be used for the quality control reagent for detecting the lymphocyte surface mark antigenicity (CD series) by adopting the cell analyzer method, the immunofluorescence method, the immunoenzyme method, a SPA (Staphylococcus aureus A protein) rosettes method and the like.
Description
Technical field
The present invention relates to the quality-control product in the Medical Immunology, relate in particular to a kind of quality-control product of drying human lymphocyte surface antigen and preparation method thereof.
Background technology
Lymphocyte plays central role in whole machine body immune response process.In peripheral blood leucocyte, lymphocyte accounts for 20%～50%[absolute value (1～3.3) * 10
9/ L], it mainly comprises T cell, B cell and NK cell.Because lymphocyte is inhomogenous cell colony, it has comprised many subgroups with different immunologic functions, therefore measure the lymphocytic cell surface mark with immunological technique, can be used for judging the immunologic function of human body, help the prevention and health care of human body and diagnosis, treatment and the prognosis of disease.
At present, the method of measuring the lymphocytic cell surface mark is a lot, but which kind of method no matter, it all is the principle of applied immunology antigen-antibody reaction, be the monoclonal antibody specific and the lymphocyte reaction of measured surface sign, show that by (as fluorescence, dyeing or garland) in different ways positive cell obtains the result.
Current; measure the monoclonal antibody (McAb) of lymphocyte CD series and worldwide carried out scale, commercialization production, different CDMcAb manufacturers, different CD McAb kind and the assay method of lymphocyte subgroup are selected according to the actual conditions of self in each laboratory.At present, because the detection method difference that each laboratory is selected for use, the experimental apparatus model differs, and Chinese lymphocyte subgroup reference value does not still have unified final conclusion, therefore in real work temporary transient with reference to the reference value in the selected separately kit instructions as criterion, thereby cause the reliability of lymphocyte subgroup testing result, comparability poor, detect difficult quality guarantee.
" national clinical examination working specification " at present (Department of Medical Administration of Ministry of Health of the People's Republic of China compiles, the third edition, publishing house of Southeast China University, 2007:349; Second edition 1997:381-384 and first published 1991:286-287) in the reagent that is used for the quality control that cellular immunity detects of regulation be to use the lymphocyte alive of liquid nitrogen cryopreservation, this reagent is selected qualified donor by trial test, get anti-freezing (heparin or ethylenediamine tetraacetic acid) blood on an empty stomach from vein, isolated lymphocytes, wash 2～3 times, be suspended in the cell freezing preservation liquid and obtain, carry out then preserving in the rearmounted liquid nitrogen of a small amount of packing, liquid nitrogen container places freezer, is used as quality control in every batch of test.During use, from liquid nitrogen container, take out a cell cryopreservation pipe, move into rapidly in 37 ℃ of water-baths and recover, the sucking-off cell suspension is centrifugal in another centrifuge tube, abandons supernatant, washs 2～3 times, add quantitative cell suspending liquid according to purposes and suspend, it is standby to be adjusted to required cell concentration.But in this technology, cell must freezing preservation in liquid nitrogen, and liquid nitrogen container must place freezer, preserves requirement condition strictness, appointed condition harshness, makes necessary expense very high; Be the cell of living because of what preserve simultaneously, in case it is improper that operating process and experiment condition have slightly, cell just is easy to damage and dead, thereby influence cellular antigens and test findings, therefore common this condition of the Chang Yinwu of clinical labororatory and ability and can't carry out this test, and the person of having ready conditions when reality is used often owing to cell separation, frozen, recovery process technology requirement height, complicated operation, waste time and energy, and the term of validity is short and be difficult to effectively be promoted.
Because lymphocyte surface antigen belongs to labile antigen, in the preservation process, very easily lose, therefore use the lymphocyte quality-control product of living, can't solve the problem of widespread usage in the real work.
Summary of the invention
Technical matters to be solved by this invention provides and a kind ofly makes lymphocyte lose biologic activity and preserve its antigen active, and keeps the breadboard different operating person's who is applicable to different people lymphocyte surface antigen detection method, different region, different condition of lymphocyte morphosis quality-control product of drying human lymphocyte surface antigen as much as possible.
Another technical matters to be solved by this invention provides the preparation method that the cryodesiccated method of a kind of usefulness makes the lymphocyte surface antigen quality-control product of drying human lymphocyte surface antigen that exists steady in a long-term.
For addressing the above problem, a kind of quality-control product of drying human lymphocyte surface antigen of the present invention is characterized in that: the cell concentration that it is made up of human lymphocyte after fixing agent is fixing and protective agent is the cell suspension of 400～6,000,000/mL; Its sign value is CD3
+: 37.2～69.1%, CD4
+: 17.9～33.7%, CD8
+: 12.4～23.9%, CD4
+/ CD8
+: 0.98～1.94.
Described fixing agent is that volumetric concentration is 0.02%～4%, the pH value is 7.0～7.4 neutral buffered formalin; This solution is meant that with distilled water and mass concentration be after 40% formaldehyde mixes by the volume ratio of 9:1, add lime carbonate, make it be hypersaturated state, promptly obtain the pH value and be 7.0, mass concentration is 4% neutral formalin solution, is that 7.0～7.4 no calcium, magnesium Han Keshi balanced salt solution is formulated by the volume ratio of 1:99～0 then with this neutral formalin solution and pH value.
Described protective agent is that mass concentration is 0.15～2.5%, the pH value is 7.0～7.4 no calcium, magnesium Han Keshi gelatin solution; This solution is that 0.15～2.5 gram gelatin is dissolved in 100mL pH value is in 7.0～7.4 no calcium, the magnesium Han Keshi balanced salt solution, and is formulated after filtering.
A kind of preparation method of aforesaid quality-control product of drying human lymphocyte surface antigen may further comprise the steps:
(1) select qualified donor by trial test, venous blood samples, adding the pH value with the volume ratio of 1:1～3 is 7.0～7.4 no calcium, magnesium Han Keshi balanced salt solution, mixing is superimposed upon on the lymphocyte separation medium, and is centrifugal;
(2) get mononuclearcell, be 7.0～7.4 no calcium, the washing of magnesium Han Keshi balanced salt solution with the pH value after, centrifugal, abandon supernatant;
(3) be that 7.0～7.4 no calcium, magnesium Han Keshi balanced salt solution are diluted to 400～6,000,000/mL cell suspension with the cell of post precipitation with the pH value, drip then in fixing agent; Under-20～30 ℃, stir and fix 0.25～48h, get the fixed cell suspension;
(4) the fixed cell suspension is filtered with nylon mesh after, centrifugal, abandon supernatant; Be to abandon supernatant after 7.0～7.4 no calcium, magnesium Han Keshi balanced salt solution or the distilled water washing with the pH value again, the fixed cell precipitation;
(5) protective agent is added in the fixed cell precipitation, is made into 400～6,000,000/mL cell suspension, be sub-packed in frozen pipe or ampere bottle with the amount of every bottle of 0.2～1.0mL after, place-80～-20 ℃ of refrigerator and cooled to freeze 4～72h at once;
(6) freeze drier is chilled in advance-38～-42 ℃, takes out frozen pipe or the ampere bottle fill cell suspension and put into freeze drier, take out behind freeze-drying 4～18h, seal immediately, and frozen pipe or ampere bottle are put-20 ℃～4 ℃ preserve down and get final product.
Venous blood in the described step (1) adopts heparin or ethylenediamine tetraacetic acid anti-coagulants to extract.
The cell suspension in the described step (3) and the volume ratio of fixing agent are 1:5～20.
Nylon mesh in the described step (4) is 200～450 orders.
The application of a kind of aforesaid quality-control product of drying human lymphocyte surface antigen in the human lymphocyte surface antigen detects, it is characterized in that: during use, in human lymphocyte surface antigen quality-control product, add in the no calcium, magnesium Han Keshi balanced salt solution, the no calcium that contains 20% newborn calf serum, magnesium Han Keshi balanced salt solution, phosphate buffer, distilled water of 100～1000 μ L any one, make its dissolving, the pH value of wherein said no calcium, magnesium Han Keshi balanced salt solution or phosphate buffer is 7.0～7.4.
The present invention compared with prior art has the following advantages:
1, because the present invention is the dried frozen aquatic products after fixing through fixing agent, be the lymphocyte that surface antigen is preserved good inanimate object activity, so antigenicity is stable, the term of validity is grown to 2～5 years, reliable in quality.
2, because the present invention be dried frozen aquatic products and sealing, and its volume, weight are little, can be lower than under 25 ℃ the normal temperature or-20 ℃～4 ℃ transportations, preservation, latter's first-selection need not special technique and condition, so can reduce storage expenses greatly.
3, the present invention does not need specific installation and condition in using, and is simple to operate, can be widely used in common clinical labororatory and carry out Quality Control work, thereby promote and improve active quality and the level that detects of human lymphocyte surface antigen.
4, product price of the present invention is cheap, is easy to get the transportation convenient storage.
5, product of the present invention can be used as methods detection lymphocytic cell surface sign (CD series) antigenic quality control reagent (reference material) such as flow cytometer method, immunofluorescence technique, immunoenzyme, SPA (staphylococcal protein A) garland method, and is used for other test items (acceptor) of relevant lymphocytic cell surface sign and the indoor and outdoor Quality Control of method; Also can be used for detecting simultaneously lymphocytic cell surface sign McAb affinity, tire, detection such as type, estimate lymphocyte surface antigen McAb.
6, as to product of the present invention through International or National Quality Control relevant speciality department---demarcate antigen active as WHO (World Health Organization (WHO)), IUIS (international immunology federation), ministry of Health of China clinical examination centralab after, International or National one-level, secondary reference material can be provided, as lymphocyte surface antigen quality control unified standard reference material, be used for the lymphocyte surface antigen quality control, estimate monoclonal antibody (McAb) quality and McAb quality control.
Embodiment
1 one kinds of quality-control product of drying human lymphocyte surface antigen of embodiment, the cell concentration that it is made up of human lymphocyte after formalin is fixing and gelatin solution is the cell suspension of 400～6,000,000/mL, is the cotton-shaped solid of canescence after freeze-drying; Adopt the flow cytometer method to measure lymphocyte subgroup (CD), obtaining its sign value is CD3
+: 37.2～69.1%, CD4
+: 17.9～33.7%, CD8
+: 12.4～23.9%, CD4
+/ CD8
+: 0.98～1.94.
The preparation method of above-mentioned quality-control product of drying human lymphocyte surface antigen is as follows:
(1) at first by 18～38 years old sex cellular immune function---lymphocyte CD 3 to healthy, non-communicable disease and other acute and chronic disease, no disease of immune system
+, CD4
+, CD8
+, CD4
+/ CD8
+Detect, the normal person of result is defined as the qualified donor of trial test.
Then the qualified donor of trial test is adopted heparin or ethylenediamine tetraacetic acid anti-coagulants venous blood samples on an empty stomach, adding the pH value with the volume ratio of 1:1～3 is 7.0～7.4 no calcium, magnesium Han Keshi balanced salt solution (no calcium, magnesium Hank ' S liquid), mixing, be superimposed upon lymphocyte separation medium (source: blood disease research institute of the Chinese Academy of Medical Sciences), with the centrifugal 15～30min of the rotating speed of 1500～2500rpm.
(2) get mononuclearcell (PBMC), be 7.0～7.4 no calcium, the washing of magnesium Hank ' S liquid 2～3 times with the pH value of 5～10 times of amounts after, with the centrifugal 5～10min of the rotating speed of 500～1500rpm, abandon supernatant.
(3) be that 7.0～7.4 no calcium, magnesium Han Keshi balanced salt solution are diluted to 400～6,000,000/mL cell suspension with the cell of post precipitation with the pH value, drip then in the pH value be 7.0～7.4, volumetric concentration is that the volume ratio of cell suspension and formalin is 1:5～20 in 0.02%～4% the buffering formalin; Fix 0.25～48h-20～30 ℃ of lower magnetic forces stirrings, get the fixed cell suspension.
Wherein formalin is meant with distilled water and mass concentration to be after 40% formaldehyde mixes by the volume ratio of 9:1, to add lime carbonate, makes it be hypersaturated state, promptly obtains the pH value and be 7.0, mass concentration is 4% neutral formalin solution; Be that 7.0～7.4 no calcium, magnesium Han Keshi balanced salt solution are formulated by the volume ratio of 1:99～0 then with this neutral formalin solution and pH value.
(4) the fixed cell suspension that step (3) is obtained filters in back immigration 5～50mL centrifuge tube with 200～450 order nylon mesh, with the centrifugal 5～10min of the rotating speed of 500～2500rpm, abandons supernatant; Be to abandon supernatant after 7.0～7.4 no calcium, magnesium Hank ' S liquid or the distilled water washing 2～3 times with the pH value again, the fixed cell precipitation.
(5) 0.15～2.5 gram gelatin being dissolved in 100mL pH value is in 7.0～7.4 no calcium, the magnesium Han Keshi balanced salt solution, with the gelatin solution that is mixed with 0.15～2.5% after the filtration of 100～200 order nylon mesh.
This gelatin solution is added in the fixed cell precipitation that step (4) obtains; use the method counting cells of blood count behind the mixing; be made into 400～6,000,000/mL cell suspension; obtain being added with the fixed cell suspension of freeze drying protectant; after being sub-packed in frozen pipe or ampere bottle with the amount of every bottle of 0.2～1.0mL then, place-80～-20 ℃ of refrigerator and cooled to freeze 4～72h at once.
(6) freeze drier is chilled in advance-38～-42 ℃, taking-up fills the frozen pipe or the ampere bottle of cell suspension and puts into freeze drier (must guard against freezing thing dissolving) rapidly, take out behind freeze-drying 4～18h, seal immediately, and frozen pipe or ampere bottle are put-20 ℃～4 ℃ preserve down and get final product.
Aforesaid operations all carries out under gnotobasis.
When embodiment 2 is applied to flow cytometer method detection lymphocyte subgroup (CD) when this quality-control product of drying human lymphocyte surface antigen, in human lymphocyte surface antigen quality-control product, add no calcium, magnesium Hank ' the S liquid of 100～1000 μ L, pH=7.0～7.4; Perhaps in quality-control product, add 100～1000 μ L, volumetric molar concentration is the phosphate buffer (PBS) of 0.01mol/L, pH=7.0～7.4; The distilled water that perhaps in quality-control product, adds 100～1000 μ L, dissolving, and then with the filtration of 250～450 purpose nylon mesh, standby.
When embodiment 3 is applied to immunofluorescence technique, AP-AAP bridging enzyme immunoassay detection lymphocyte subgroup (CD) when this quality-control product of drying human lymphocyte surface antigen, in human lymphocyte surface antigen quality-control product, add the no calcium that contains 20% newborn calf serum, magnesium Hank ' the S liquid of 100～1000 μ L, pH=7.0～7.4; Perhaps in quality-control product, add 100～1000 μ L, volumetric molar concentration is the PBS that contains 20% newborn calf serum of 0.01mol/L, pH=7.0～7.4; The distilled water that contains 20% newborn calf serum that perhaps adds 100～1000 μ L in quality-control product, the dissolving back is standby.
Embodiment 4 is when this quality-control product of drying human lymphocyte surface antigen is applied to monoclonal antibody SPA garland method (McAb-A-E direct method or indirect method) when detecting lymphocyte subgroup (CD), adds 100～1000 μ L, pH value and be 7.0～7.4 no calcium, magnesium Hank ' S liquid in human lymphocyte surface antigen quality-control product; The distilled water that perhaps adds 100～1000 μ L in quality-control product, the dissolving back is standby.
Should be appreciated that embodiment discussed here and embodiment can propose various improvement and variation just in order to illustrate to the people who is familiar with this field, these improvement and variation will be included in the application's spirit and scope and the appended claim scope.
Claims (8)
1. quality-control product of drying human lymphocyte surface antigen is characterized in that: the cell concentration that it is made up of human lymphocyte after fixing agent is fixing and protective agent is the cell suspension of 400～6,000,000/mL; Its sign value is CD3
+: 37.2～69.1%, CD4
+: 17.9～33.7%, CD8
+: 12.4～23.9%, CD4
+/ CD8
+: 0.98～1.94.
2. a kind of quality-control product of drying human lymphocyte surface antigen as claimed in claim 1 is characterized in that: described fixing agent is that volumetric concentration is 0.02%～4%, the pH value is 7.0～7.4 neutral buffered formalin; This solution is meant that with distilled water and mass concentration be after 40% formaldehyde mixes by the volume ratio of 9:1, add lime carbonate, make it be hypersaturated state, promptly obtain the pH value and be 7.0, mass concentration is 4% neutral formalin solution, is that 7.0～7.4 no calcium, magnesium Han Keshi balanced salt solution is formulated by the volume ratio of 1:99～0 then with this neutral formalin solution and pH value.
3. a kind of quality-control product of drying human lymphocyte surface antigen as claimed in claim 1 is characterized in that: described protective agent is that mass concentration is 0.15～2.5%, the pH value is 7.0～7.4 no calcium, magnesium Han Keshi gelatin solution; This solution is that 0.15～2.5 gram gelatin is dissolved in 100mL pH value is in 7.0～7.4 no calcium, the magnesium Han Keshi balanced salt solution, and is formulated after filtering.
4. the preparation method of a quality-control product of drying human lymphocyte surface antigen as claimed in claim 1 may further comprise the steps:
(1) select qualified donor by trial test, venous blood samples, adding the pH value with the volume ratio of 1:1～3 is 7.0～7.4 no calcium, magnesium Han Keshi balanced salt solution, mixing is superimposed upon on the lymphocyte separation medium, and is centrifugal;
(2) get mononuclearcell, be 7.0～7.4 no calcium, the washing of magnesium Han Keshi balanced salt solution with the pH value after, centrifugal, abandon supernatant;
(3) be that 7.0～7.4 no calcium, magnesium Han Keshi balanced salt solution are diluted to 400～6,000,000/mL cell suspension with the cell of post precipitation with the pH value, drip then in fixing agent; Under-20～30 ℃, stir and fix 0.25～48h, get the fixed cell suspension;
(4) the fixed cell suspension is filtered with nylon mesh after, centrifugal, abandon supernatant; Be to abandon supernatant after 7.0～7.4 no calcium, magnesium Han Keshi balanced salt solution or the distilled water washing with the pH value again, the fixed cell precipitation;
(5) protective agent is added in the fixed cell precipitation, is made into 400～6,000,000/mL cell suspension, be sub-packed in frozen pipe or ampere bottle with the amount of every bottle of 0.2～1.0mL after, place-80～-20 ℃ of refrigerator and cooled to freeze 4～72h at once;
(6) freeze drier is chilled in advance-38～-42 ℃, takes out frozen pipe or the ampere bottle fill cell suspension and put into freeze drier, take out behind freeze-drying 4～18h, seal immediately, and frozen pipe or ampere bottle are put-20 ℃～4 ℃ preserve down and get final product.
5. the preparation method of a kind of quality-control product of drying human lymphocyte surface antigen as claimed in claim 4 is characterized in that: venous blood employing heparin in the described step (1) or the extraction of ethylenediamine tetraacetic acid anti-coagulants.
6. the preparation method of a kind of quality-control product of drying human lymphocyte surface antigen as claimed in claim 4, it is characterized in that: the cell suspension in the described step (3) and the volume ratio of fixing agent are 1:5～20.
7. the preparation method of a kind of quality-control product of drying human lymphocyte surface antigen as claimed in claim 4, it is characterized in that: the nylon mesh in the described step (4) is 200～450 orders.
8. the application of quality-control product of drying human lymphocyte surface antigen as claimed in claim 1 in the human lymphocyte surface antigen detects, it is characterized in that: during use, in human lymphocyte surface antigen quality-control product, add in the no calcium, magnesium Han Keshi balanced salt solution, the no calcium that contains 20% newborn calf serum, magnesium Han Keshi balanced salt solution, phosphate buffer, distilled water of 100～1000 μ L any one, make its dissolving, the pH value of wherein said no calcium, magnesium Han Keshi balanced salt solution or phosphate buffer is 7.0～7.4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810150909A CN101363846B (en) | 2008-09-05 | 2008-09-05 | Quality-control product of freeze-drying human lymphocyte surface antigen and method for making same |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810150909A CN101363846B (en) | 2008-09-05 | 2008-09-05 | Quality-control product of freeze-drying human lymphocyte surface antigen and method for making same |
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| Publication Number | Publication Date |
|---|---|
| CN101363846A true CN101363846A (en) | 2009-02-11 |
| CN101363846B CN101363846B (en) | 2012-10-10 |
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|---|---|---|---|
| CN200810150909A Expired - Fee Related CN101363846B (en) | 2008-09-05 | 2008-09-05 | Quality-control product of freeze-drying human lymphocyte surface antigen and method for making same |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012038041A3 (en) * | 2010-09-21 | 2012-05-18 | Herz- Und Diabeteszentrum Nordrhein-Westfalen Krankenhausbetriebsgesellschaft Bad Oeynhausen Mbh | Stabilized leukocytes and their use in hiv-diagnosis and therapy |
| CN107576790A (en) * | 2017-11-03 | 2018-01-12 | 深圳市巴德生物科技有限公司 | A kind of drying human lymphocyte CD4 surface antigen quality-control products and preparation method thereof |
| CN107576788A (en) * | 2017-11-03 | 2018-01-12 | 深圳市巴德生物科技有限公司 | A kind of reference product for detecting leukocyte differentiation antigen and preparation method thereof |
| CN111351930A (en) * | 2020-02-20 | 2020-06-30 | 泛肽生物科技(浙江)有限公司 | Quality control product of active human lymphocyte surface antigen and preparation method thereof |
| CN111528219A (en) * | 2020-05-13 | 2020-08-14 | 上海市计量测试技术研究院 | Freeze-drying protective agent for T lymphocyte subpopulation counting standard substance and application thereof |
| CN114152739A (en) * | 2021-12-08 | 2022-03-08 | 江西赛基生物技术有限公司 | Fluorescent antibody freeze-dried pellet and preparation method thereof |
-
2008
- 2008-09-05 CN CN200810150909A patent/CN101363846B/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012038041A3 (en) * | 2010-09-21 | 2012-05-18 | Herz- Und Diabeteszentrum Nordrhein-Westfalen Krankenhausbetriebsgesellschaft Bad Oeynhausen Mbh | Stabilized leukocytes and their use in hiv-diagnosis and therapy |
| US8900864B2 (en) | 2010-09-21 | 2014-12-02 | Nordrhein-Wesfalen Krankenhausbetriebsgesellschaft Bad Oeynhausen mbH | Stabilized leukocytes and their use |
| CN107576790A (en) * | 2017-11-03 | 2018-01-12 | 深圳市巴德生物科技有限公司 | A kind of drying human lymphocyte CD4 surface antigen quality-control products and preparation method thereof |
| CN107576788A (en) * | 2017-11-03 | 2018-01-12 | 深圳市巴德生物科技有限公司 | A kind of reference product for detecting leukocyte differentiation antigen and preparation method thereof |
| CN111351930A (en) * | 2020-02-20 | 2020-06-30 | 泛肽生物科技(浙江)有限公司 | Quality control product of active human lymphocyte surface antigen and preparation method thereof |
| CN111528219A (en) * | 2020-05-13 | 2020-08-14 | 上海市计量测试技术研究院 | Freeze-drying protective agent for T lymphocyte subpopulation counting standard substance and application thereof |
| CN111528219B (en) * | 2020-05-13 | 2022-03-15 | 上海市计量测试技术研究院 | Freeze-drying protective agent for T lymphocyte subpopulation counting standard substance and application thereof |
| CN114152739A (en) * | 2021-12-08 | 2022-03-08 | 江西赛基生物技术有限公司 | Fluorescent antibody freeze-dried pellet and preparation method thereof |
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| CN101363846B (en) | 2012-10-10 |
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