CN101178409A - Monoclonal antibody IgM type RhD blood type shaped reagent - Google Patents
Monoclonal antibody IgM type RhD blood type shaped reagent Download PDFInfo
- Publication number
- CN101178409A CN101178409A CNA2006101180257A CN200610118025A CN101178409A CN 101178409 A CN101178409 A CN 101178409A CN A2006101180257 A CNA2006101180257 A CN A2006101180257A CN 200610118025 A CN200610118025 A CN 200610118025A CN 101178409 A CN101178409 A CN 101178409A
- Authority
- CN
- China
- Prior art keywords
- monoclonal
- monoclonal cell
- cell system
- concentrate
- monoclonal antibody
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention provides an RhD blood grouping reagent with a monoclonal antibody IgM type, which consists of anti-D monoclonal cell culture concentrated solution and dilution solution, the anti-D monoclonal cell culture concentrated solution is prepared by using the following method, firstly, the invention includes an establishing process of a hybrid cell system, the establishing process of the hybrid cell system includes a process of making use of lymph cells transformed by EB virus to hybridize with the mouse bone marrow cells, the hybrid monoclonal cell system is established by the process, secondly, the invention includes a culture process of the hybrid monoclonal cell system, the culture solution of the monoclonal cells is collected to carry out the concentration by the culture of the hybrid monoclonal cell system, so as to obtain the anti-D monoclonal cell culture concentrated solution; the dilution solution consists of sodium chloride, sodium phosphate with 12 water molecules, potassium dihydrogen phosphate, stabilizer, sodium azide, Tween 20 and distilled water; as the monoclonal antibody is adopted, the invention has strong specific antibody response and high accuracy rate of the detection.
Description
Technical field:
The present invention relates to biochemical field, relate in particular to the reagent of identifying blood group, is a kind of RhD blood grouping reagent of monoclonal antibody IgM type specifically.
Background technology:
General clinically according to the having or not of erythrocyte surface D antigen, the Rh blood group is divided into the positive and Rh feminine gender of Rh.The negative mother of Rh is when having bred the positive fetus of Rh, and its parent can produce Rh antibody, can cause neonatal hemolytic disease.Therefore the detection of the Rh positive is even more important for this disease.
Take from people's serum for the antibody of surveyor's erythrocyte blood type clinically now more, so, not only consume a large amount of human serums, in addition some that may exist in Shang Keyin human serum infections sources of understanding and cause unthinkable consequence not as yet.And how to detect the RhD blood group clinically at present with polyclonal antibody, for its antibody response poor specificity of polyclonal antibody, False Rate is higher.
Summary of the invention:
The object of the present invention is to provide a kind of RhD blood grouping reagent of monoclonal antibody IgM type, the antibody that described monoclonal antibody IgM type RhD blood grouping reagent will solve surveyor's erythrocyte blood type of the prior art is taken from people's serum more, need to consume a large amount of human serums, and some infection sources technical matters that can cause the people to infect of understanding not as yet that may exist in the human serum, solved simultaneously that the polyclonal antibody atopic is poor, the False Rate technical problems of high.
The RhD blood grouping reagent of this monoclonal antibody IgM type of the present invention, form by anti-D monoclonal cell cultivation concentrate and dilution, wherein, described anti-D monoclonal cell is cultivated concentrate and is adopted following method preparation, at first comprise a process of setting up hybrid cell line, the described process of setting up hybrid cell line comprises that one is utilized the lymphocyte of Epstein-Barr virus conversion and the process of mouse bone marrow cells hybridization, set up the monoclonal cell system of hybridization by this process, next comprises a process of cultivating the monoclonal cell system of hybridization, the cultivation of the monoclonal cell system by hybridization, collect the nutrient solution of monoclonal cell, concentrate, the monoclonal cell that obtains anti-D is cultivated concentrate, described dilution is by sodium chloride, sodium hydrogen phosphate with 12 hydrones, potassium dihydrogen phosphate, stabilizing agent, Sodium azide and polysorbas20 and distilled water are formed, with every 10L dilution metering, the sodium chloride that wherein contains 170-190g, the sodium hydrogen phosphate of 90-100g with 12 hydrones, the potassium dihydrogen phosphate of 30-35g, the stabilizing agent of 480-520g, the Sodium azide of 8-12g and the polysorbas20 of 0.8-1.3ml, surplus are distilled water.
Further, in described dilution, with every 10L dilution metering, wherein contain the sodium chloride of 180g, the sodium hydrogen phosphate of 95g, the potassium dihydrogen phosphate of 33g, the stabilizing agent of 500g, the Sodium azide of 10g and the polysorbas20 of 1.0ml with 12 hydrones, surplus is a distilled water.
Further, the monoclonal cell of described hybridization system adopts the RUM-1 clone of Britain Serologicals company.
Further, described monoclonal cell is cultivated the FFMU concentrate that concentrate adopts Britain Serologicals company.
Principle of work of the present invention is: utilize monoclonal antibody, mix with the human erythrocyte of certain volume, on glass sheet or other carrier aggregation takes place, according to the cohesion result, think and judge the blood type.If erythrocyte surface has D antigen, do the time spent with the present invention, specific antigen-antibody reaction will take place, make red born of the same parents that aggegation take place, be the Rh positive.If aggegation does not take place, show that erythrocyte surface does not have D antigen, be the Rh feminine gender.
The present invention and prior art contrast, and its effect is actively with tangible.The present invention is because adopted monoclonal antibody to be used for the evaluation of human red blood cell blood group, saved a large amount of human serums, avoided the infectious agent that may exist in the human serum, simultaneously monoclonal antibody of the present invention has that height is tired and specificity, has improved the accuracy rate height that detects.
Embodiment:
Embodiment 1
The RhD blood grouping reagent of a kind of monoclonal antibody IgM type of the present invention, the IgM type RhD blood grouping reagent of described monoclonal antibody has solved polyclonal antibody liquid injection of the prior art owing to its antibody response poor specificity, False Rate technical problems of high.A kind of RhD blood grouping reagent of monoclonal antibody IgM type, form by anti-D monoclonal cell cultivation concentrate and dilution, wherein, described anti-D monoclonal cell is cultivated concentrate and is adopted following method preparation, at first comprise a process of setting up hybrid cell line, the described process of setting up hybrid cell line comprises that one is utilized the lymphocyte of Epstein-Barr virus conversion and the process of mouse bone marrow cells hybridization, set up the monoclonal cell system of hybridization by this process, next comprises a process of cultivating the monoclonal cell system of hybridization, the cultivation of the monoclonal cell system by hybridization, collect the nutrient solution of monoclonal cell, concentrate, the monoclonal cell that obtains anti-D is cultivated concentrate, described dilution is by sodium chloride, sodium hydrogen phosphate with 12 hydrones, potassium dihydrogen phosphate, stabilizing agent, Sodium azide and polysorbas20 and distilled water are formed, with every 10L dilution metering, the sodium chloride that wherein contains 180g, the sodium hydrogen phosphate of 95g with 12 hydrones, the potassium dihydrogen phosphate of 33g, the stabilizing agent of 500g, the Sodium azide of 10g and the polysorbas20 of 1.0ml, surplus are distilled water.
Further, the monoclonal cell of described hybridization system adopts the RUM-1 clone of Britain Serologicals company.
Further, described monoclonal cell is cultivated the FFMU concentrate that concentrate adopts Britain Serologicals company.
Embodiment 2
Flat band method detects the RhD blood group
1. add an anti-D reagent on slide;
2. the tested red cell suspension that adds a 35-45% again, this red cell suspension can be the salt aqueous suspensions, also can be the red blood cells that is suspended in self blood plasma or the serum;
3. circular again slide zone mixing, slide is slowly rocked in front and back, naked eyes sentence read result about 2 minutes;
4. as the arnotto born of the same parents aggegation taking place, is the Rh positive.If aggegation does not take place, show that erythrocyte surface does not have D antigen, be the Rh feminine gender.
5. if negative, can detect again once, increase reliability.
Embodiment 3
Test tube method detects the RhD blood group
1. add an anti-D reagent in a test tube that mark is good in advance;
2. the tested red cell suspension that adds an about 3-5% again in test tube, mixing;
1. centrifugal, speed and time can select following any one:
Rotating speed 1000RPM, 1 minute time;
Rotating speed 3400RPM, 15 seconds time;
4. whether check haemolysis (haemolysis may be positive findings, or bacterial contamination), jiggle then, cell is suspended again;
5. observe the aggegation situation, and the immediate record result.
6. as the arnotto born of the same parents aggegation taking place, is the Rh positive.If aggegation does not take place, show that erythrocyte surface does not have D antigen, be the Rh feminine gender.
7. if negative, can detect again once, increase reliability.
Embodiment 4
The micro plate method detects the RhD blood group
1. add an anti-D reagent in the micropore of micro plate;
2. add the tested red cell suspension of an about 3-5% again in micropore, cell suspension needs prepared beforehand in physiological saline;
3. use the method for machinery with the micro plate mixing;
4. centrifugal with board-like hydro-extractor: 2000RPM, 30 seconds time;
5. whether check haemolysis (haemolysis may be positive findings, or bacterial contamination);
6. use the mechanical oscillation micro plate, suspension cell is judged aggegation according to the suspension situation again, and red cell agglutination appears in positive reaction, and negative reaction then cell suspension can occur in micropore.
7. if negative, can detect again once, increase reliability.
Claims (4)
1. the RhD blood grouping reagent of a monoclonal antibody IgM type, form by anti-D monoclonal cell cultivation concentrate and dilution, wherein, described anti-D monoclonal cell is cultivated concentrate and is prepared by the following method, at first comprise a process of setting up hybrid cell line, the described process of setting up hybrid cell line comprises that one is utilized the lymphocyte of Epstein-Barr virus conversion and the process of mouse bone marrow cells hybridization, set up the monoclonal cell system of hybridization by this process, next comprises a process of cultivating the monoclonal cell system of hybridization, the cultivation of the monoclonal cell system by hybridization, collect the nutrient solution of monoclonal cell, concentrate, the monoclonal cell that obtains anti-D is cultivated concentrate, it is characterized in that: described dilution is by sodium chloride, sodium hydrogen phosphate with 12 hydrones, potassium dihydrogen phosphate, stabilizing agent, Sodium azide and polysorbas20 and distilled water are formed, with every 10L dilution metering, the sodium chloride that wherein contains 170-190g, the sodium hydrogen phosphate of 90-100g with 12 hydrones, the potassium dihydrogen phosphate of 30-35g, the stabilizing agent of 480-520g, the Sodium azide of 8-12g and the polysorbas20 of 0.8-1.3ml, surplus are distilled water.
2. the IgM type RhD blood grouping reagent of monoclonal antibody as claimed in claim 1, it is characterized in that: in described dilution, with every 10L dilution metering, wherein contain the sodium chloride of 180g, the sodium hydrogen phosphate with 12 hydrones of 95g, the potassium dihydrogen phosphate of 33g, the stabilizing agent of 500g, the Sodium azide of 10g and the polysorbas20 of 1.0ml, surplus is a distilled water.
3. the IgM type RhD blood grouping reagent of monoclonal antibody as claimed in claim 1 is characterized in that: the RUM-1 clone of the monoclonal cell system employing Britain Serologicals company of described hybridization.
4. the IgM type RhD blood grouping reagent of monoclonal antibody as claimed in claim 1 is characterized in that: described monoclonal cell is cultivated the FFMU concentrate that concentrate adopts Britain Serologicals company.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006101180257A CN101178409A (en) | 2006-11-07 | 2006-11-07 | Monoclonal antibody IgM type RhD blood type shaped reagent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006101180257A CN101178409A (en) | 2006-11-07 | 2006-11-07 | Monoclonal antibody IgM type RhD blood type shaped reagent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101178409A true CN101178409A (en) | 2008-05-14 |
Family
ID=39404742
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2006101180257A Pending CN101178409A (en) | 2006-11-07 | 2006-11-07 | Monoclonal antibody IgM type RhD blood type shaped reagent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101178409A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102445550A (en) * | 2010-10-09 | 2012-05-09 | 苏州苏大赛尔免疫生物技术有限公司 | ABO, RhD blood typing reagent card, preparation method thereof and antibody diluting solution adopted in preparation method |
| CN101701961B (en) * | 2009-11-25 | 2013-03-06 | 江阴力博医药生物技术有限公司 | Method for preparing typing detection reagent card of blood types of A, B and O |
| CN106267405B (en) * | 2016-07-01 | 2019-02-19 | 翁炳焕 | Female tire blood group incompatibility haemolysis disease therapeutic apparatus |
| WO2023143629A1 (en) * | 2022-01-26 | 2023-08-03 | 江苏力博医药生物技术股份有限公司 | Fully-humanized anti-human erythrocyte rhd whole-molecule igg, and preparation method therefor and use thereof |
-
2006
- 2006-11-07 CN CNA2006101180257A patent/CN101178409A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101701961B (en) * | 2009-11-25 | 2013-03-06 | 江阴力博医药生物技术有限公司 | Method for preparing typing detection reagent card of blood types of A, B and O |
| CN102445550A (en) * | 2010-10-09 | 2012-05-09 | 苏州苏大赛尔免疫生物技术有限公司 | ABO, RhD blood typing reagent card, preparation method thereof and antibody diluting solution adopted in preparation method |
| CN102445550B (en) * | 2010-10-09 | 2014-02-05 | 苏州苏大赛尔免疫生物技术有限公司 | ABO, RhD blood typing reagent card, preparation method thereof |
| CN106267405B (en) * | 2016-07-01 | 2019-02-19 | 翁炳焕 | Female tire blood group incompatibility haemolysis disease therapeutic apparatus |
| WO2023143629A1 (en) * | 2022-01-26 | 2023-08-03 | 江苏力博医药生物技术股份有限公司 | Fully-humanized anti-human erythrocyte rhd whole-molecule igg, and preparation method therefor and use thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Nébor et al. | Fetal hemoglobin and hydroxycarbamide moduate both plasma concentration and cellular origin of circulating microparticles in sickle cell anemia children | |
| CN103336110B (en) | Whole blood quality control material and preparation method thereof | |
| CN102445550B (en) | ABO, RhD blood typing reagent card, preparation method thereof | |
| CN111551712A (en) | Novel test strip and kit for quickly detecting coronavirus IgM/IgG two-in-one and preparation method of test strip and kit | |
| CN102692507A (en) | Kit for measuring lipoprotein-associated phospholipase A2 (LPPLA2) (by adopting latex enhanced turbidimetric immunoassay) | |
| CN108680756A (en) | A kind of incomplete antibody detection kit and detection method | |
| CN101178409A (en) | Monoclonal antibody IgM type RhD blood type shaped reagent | |
| CN103344772A (en) | Novel Miltenberger blood group antibody detecting method | |
| CN104965089A (en) | Novel platelet antibody kit using micro-column gel technique and preparing method thereof | |
| CN102507961B (en) | Newborn ABO/Rh blood grouping reagent card and preparation method thereof | |
| CN103063845A (en) | Nanometer magnetic particle chemiluminiscence kit, preparation method and detection method of hepatitis B virus surface-antibody | |
| CN203083997U (en) | ABO and RhD blood group antigen detection card | |
| CN102692516B (en) | Lupus anticoagulant (LA) screening and determining reagent kit (freezing method) | |
| CN102879560A (en) | Method for accurately and rapidly detecting antigens and antibodies in human serum in trace mode | |
| CN103602632A (en) | Method for separating fetal nucleated red blood cells in maternal peripheral blood | |
| CN104818280A (en) | Single-stranded aptamer and application thereof | |
| CN113252914B (en) | Antibody diluent for Rh system parting detection card and detection card | |
| CN102798718A (en) | Previous S1 antigen detection kit for hepatitis B virus and preparation method of previous S1 antigen detection kit | |
| CN1206537C (en) | Reagent red blood cell | |
| CN102445549A (en) | Novel column agglutination glass microsphere clamping type blood type detection reagent | |
| CN105784868A (en) | Method for detecting lactulose in milk | |
| CN103323586A (en) | Blood platelet magnetizing and immunolabeling analysis method | |
| CN104730231B (en) | A kind of sample buffer for fluorescence immunoassay detection by quantitative and application thereof | |
| CN105092863B (en) | In-vitro hemolytic test blood matching method | |
| CN106771134B (en) | A kind of carp herpesvirusⅡtype sandwich ELISA detection kit and detection method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20080514 |