CN101644704A - Preparation method for rheumatoid factor detection reagent - Google Patents
Preparation method for rheumatoid factor detection reagent Download PDFInfo
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- CN101644704A CN101644704A CN200910092374A CN200910092374A CN101644704A CN 101644704 A CN101644704 A CN 101644704A CN 200910092374 A CN200910092374 A CN 200910092374A CN 200910092374 A CN200910092374 A CN 200910092374A CN 101644704 A CN101644704 A CN 101644704A
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Abstract
The invention provides a preparation method for rheumatoid factor detection reagent. The method comprises the following steps of: coupling IgG molecules and latex particles to obtain IgG sensitizing latex; digesting and decomposing obtained IgG sensitizing latex by enzyme to obtain mixture of Fab fragment and Fc fragment sensitizing latex; separating and purifying the Fab fragment and Fc fragmentsensitizing latex to obtain the Fc fragment sensitizing latex; causing the Fc fragment on the sensitizing latex to be denatured, and obtaining the denatured Fc fragment sensitizing latex which is prepared into rheumatoid factor detection reagent. The method not only can produce rheumatoid factor detection reagent with high specificity, but also has low production cost, simple technique and is convenient to large-scale industrialization production.
Description
Technical field
The present invention relates to the preparation method of rheumatoid factor (RF) detectable, described detectable is used for measuring the content of serum rheumatoid factor.
Background technology
Rheumatoid factor (RF) is the specific antibody of antigenic determinant on the antagonism human body IgG molecule Fc fragment, is the autoantibody of anti-IgG, increases in the serum to be common in rheumatoid arthritis.The rheumatoid factor detection reagent box is used for measuring the content of serum rheumatoid factor clinically.At present, the detection method that can be used for the rheumatoid factor kit of Biochemical Analyzer roughly is divided into two kinds according to measuring principle: a kind of is to produce turbidity with the antibody of anti-RF and its reaction to detect, it is immunoturbidimetry, for example, there is the rheumatoid factor kit of Britain's Landau (Randox) company etc. the commercially available prod that utilizes this method to detect at present; Another kind is the antigen that utilizes the RF correspondence, i.e. sex change IgG and its reaction detects, for example, and the rheumatoid factor detection reagent that the commercially available prod that utilizes this method to detect at present has and light (Wako) company produces etc.Though the rheumatoid factor detection reagent atopic height that preceding a kind of method obtains, the problem that exists is that sensitivity is lower.Though the rheumatoid factor detection reagent that a kind of method in back obtains is highly sensitive, a major issue that exists is that atopic is poor, can not guarantee that promptly used IgG all is the IgG of sex change.At a kind of method in back, in order to improve the specificity of reaction, after in the past technology adopts the IgG fragmentation is handled, the method that the Fab section is separated with the Fc section, wherein, the Fab section be separating of Fc section to prepare to have the key of high specific RF antigen.At present, Fab and Fc section separation method commonly used is affinity chromatography, gel filtration, ion-exchange, electrophoresis etc.For example, people such as Coleman L. utilize gradient electrophoresis that Fab is separated (referring to non-patent literature 1) with the Fc fragment, but, this method need prepare electrophoresis equipment, pH and ionic strength to separation system are had relatively high expectations, and it is limited to handle sample size, has operating process length consuming time, and cost is higher, is difficult to use in the problem that large-scale industrialization is produced.
At present, need a kind of can be fast and convenient and need not spend the method for expensive rheumatoid factor detection reagent that just can large-scale industrialization production high specific.
Non-patent literature 1:Purification of Fab fragments from a monoclonal antibodypapain digest by Gradiflow electrophoresis, Coleman L.Mahler SM.ProteinExpression and Purification.2003 Vol.32 (No.2)
Summary of the invention
The object of the present invention is to provide a kind of preparation method of rheumatoid factor detection reagent, it is low and be convenient to the advantage of large-scale production that it can take into account atopic height, the production cost of rheumatoid factor detection reagent.
In order to solve the problems of the technologies described above, provide following technical scheme among the present invention.
1. the preparation method of rheumatoid factor detection reagent may further comprise the steps:
With IgG molecule and latex particle coupling, obtain the IgG sensitizing latex;
The IgG sensitizing latex that obtains is decomposed with enzymic digestion, obtain the potpourri of Fab fragment and Fc fragment sensitizing latex;
The Fab fragment is separated with the Fc fragment sensitizing latex, obtain the Fc fragment sensitizing latex;
Make Fc fragment sex change in the Fc fragment sensitizing latex, obtain the Fc fragment sensitizing latex of sex change, make rheumatoid factor detection reagent.
2. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, the step of IgG molecule and latex particle coupling is carried out under stirring condition in damping fluid, the buffering range of described damping fluid is at pH6.0-8.5, buffer concentration is 20-100mM, and the stirring revolution is 60-200rpm.
3. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, the step of IgG molecule and latex particle coupling is carried out under 10-40 ℃ temperature.
4. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, described latex particle is the physisorption microballoon, and the particle diameter of latex particle is in the scope of 100-200nm.
5. according to the preparation method of the rheumatoid factor detection reagent of claim 4 record, wherein, the particle diameter of described latex particle is in the scope of 110-180nm.
6. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, the purity of IgG molecule is more than 95%.
7. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, the digestion temperature of IgG molecule sensitizing latex is 30-40 ℃.
8. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, the digestion time of IgG molecule sensitizing latex is 4-5 hour.
9. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, described enzyme is a papain.
10. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, separate with the Fc fragment sensitizing latex by the centrifugal Fab of making fragment, centrifugal acceleration is 10000-20000g.
By method of the present invention, not only can produce the rheumatoid factor detection reagent of high specific, and production cost is low, technology is simple, is convenient to large-scale industrial production.
Description of drawings
Fig. 1 is the preparation method's of the rheumatoid factor detection reagent of the present invention synoptic diagram of step, 1 expression latex particle wherein, 2 expression IgG molecules, 3 expression Fab fragments, 4 expression Fc fragment sensitizing latex.
Fig. 2 represents the Fc fragment purity checking result of the detectable of the present invention of preparation in the preparation example 1, and wherein, 1 is that (unit: KD), 2 represent the Fc fragment of preparation examples 1 preparation to albumen relative molecular mass marker, 3 expression reference substances 1 (available from Calbiochem company).
Fig. 3 is the detectable of the present invention of preparation in the expression preparation example 1 of the present invention and the figure that reference substance 3 is measured the clinical sample correlativity.The transverse axis and the longitudinal axis represent to use the RF detectable of preparation example 1 preparation of the present invention and the content that reference substance reagent detects rheumatoid factor in the sample, unit (U/m1) respectively.
Fig. 4 is the figure that expression reference substance 2 and reference substance 3 are measured the clinical sample correlativity.The transverse axis and the longitudinal axis represent to use reference substance 2 and reference substance 3 to detect the content of the rheumatoid factor in the sample, unit (U/ml) respectively.
Embodiment
Below specify technical scheme of the present invention, but the invention is not restricted to following concrete embodiment.
The preparation method of rheumatoid factor detection reagent of the present invention may further comprise the steps: with IgG molecule and latex particle coupling, obtain IgG molecule sensitizing latex; The IgG molecule sensitizing latex that obtains is decomposed with enzymic digestion, obtain the potpourri of Fab fragment and Fc fragment sensitizing latex; The Fab fragment is separated with the Fc fragment sensitizing latex, obtain the Fc fragment sensitizing latex; Make the sex change of Fc fragment sensitizing latex hot polymerization, obtain the Fc fragment sensitizing latex of sex change, make rheumatoid factor detection reagent.
Above-mentioned sex change can adopt conventional method to carry out, and is not limited to the method for hot polymerization sex change.
In addition, also can with the Fab fragment with after the Fc fragment sensitizing latex is separated, further carry out purifying.
Among the preparation method of rheumatoid factor detection reagent of the present invention, the inventory of IgG molecule is calculated to the theoretical maximum connection amount that connects the IgG molecule of latex particle by 10 times.
Among the preparation method of rheumatoid factor detection reagent of the present invention, the step of IgG molecule and latex particle coupling is preferably carried out in damping fluid, the buffering range of employed damping fluid is pH6.0-8.5, be preferably pH7.0-8.0, pH7.3-7.5 more preferably, can use PBS damping fluid or GOOD damping fluid, 2-(N-morpholino) ethyl sulfonic acid (MES) damping fluid for example, N, N-bicine N-(Bicine) damping fluid, 4-hydroxyethyl piperazine ethanesulfonic acid (HEPES) damping fluid etc., buffer concentration can be 20-100mM, be preferably 30-60mM, more preferably 40-50mM; The step of IgG molecule and latex particle coupling is preferably carried out under stirring condition, and the stirring revolution is 60-200rpm, is preferably 80-150rpm, more preferably 90-110rpm.
Among the preparation method of rheumatoid factor detection reagent of the present invention, the step of IgG molecule and latex particle coupling can be carried out under 10-40 ℃ temperature, and preferred 15-35 ℃, more preferably 20-25 ℃.
Among the preparation method of rheumatoid factor detection reagent of the present invention, described latex particle is the physisorption microballoon, be preferably the latex of polymethylmethacrylate, polystyrene or both multipolymers, polystyrene latex more preferably, the particle diameter of latex particle is in the scope of 100-200nm, be preferably 110-190nm, 110-180nm more preferably, more preferably 120-170nm further is preferably 125-165nm, especially be preferably 130-160nm, be preferably 140-155nm especially.
Above-mentioned particle diameter is to use differential mobility analysis instrument (Differential Mobility Analyzer, DMA), the mean grain size that records of device such as transmission electron microscope or optical microscope, assay method is the method that adopts this area commonly used, for example is the method for regulations such as Japanese national industrial technology synthetic study institute, USA National Institute of Standard and Technology, International Bureau of Wieghts and Measurements.
Among the preparation method of rheumatoid factor detection reagent of the present invention, the purity of IgG molecule is more than 95%, and preferred purity is greater than 98%.
Among the preparation method of rheumatoid factor detection reagent of the present invention, the digestion temperature of IgG molecule sensitizing latex is 30-40 ℃, preferred 33-38 ℃, and more preferably 35-37 ℃; The digestion time of IgG molecule sensitizing latex is 4-5 hour, preferred 4-4.6 hour, and more preferably 4.3-4.5 hour; Described enzyme is a papain.
Among the preparation method of rheumatoid factor detection reagent of the present invention, separate with the Fc fragment sensitizing latex by the centrifugal Fab of making fragment, centrifugal acceleration is 10000-20000g, preferred 12000-15000g.For the Fc fragment is further purified, can carry out repeatedly centrifugally repeatedly, clean.
Among the preparation method of rheumatoid factor detection reagent of the present invention, be to guarantee the latex particle full and uniform dispersion, can adopt vortex vibration, ultrasonic method such as resuspended; Used ultrasonic condition for example can be 100-300w during latex disperseed, 5-10min, ice-water bath.
Among the preparation method of rheumatoid factor detection reagent of the present invention, can prepare the Fc fragment sensitizing latex of high specificity by the hot polymerization sex change, the hot polymerization condition is 60 ℃ of effects 30 minutes.
Embodiment
The invention is further illustrated by the following examples, but it should be interpreted as limitation of the present invention.
[experiment material]
Latex carrier (manufacturing of JSR company);
Papain is (available from MERCK company, CB5125);
Human IgG (available from Chinese Military Medical Science Institute);
Reference substance 1 (human IgG Fc fragment (Human IgG Fc Fragment Plasma (trade name), Calbiochem company makes);
Reference substance 2 (the Fc fragment of reference substance 1 and latex particle coupling, sex change and obtain RF detectable);
Reference substance 3 (RF detection kit (RHEUMATOID FACTOR Reagent kit (trade name)), Randox company makes, the RF detection kit of immunoturbidimetry, detecting principle is with anti-human rheumatoid factor antibody and RF immune response to take place to detect);
Hydro-extractor (the freezing desk centrifuge of Beckman Allegra 64R high speed)
Preparation example 1
(A) coupling of IgG and latex particle
1) takes by weighing the 10.6mg human IgG,, make solution 1 with the PBS damping fluid 5ml dissolving of 100mM;
2) get the polymethylmethacrylate latex of 0.1g particle diameter 135nm 10% (content of latex particle), fixed molten with the PBS damping fluid of 50mM to 5ml, make solution 2;
3) solution 2 is slowly joined in the solution 1,40 ℃ of bath temperatures slowly stirred (rotating speed 100rpm) 2 hours;
4) 10000g after the PBS damping fluid 10ml cleaning with 50mM, uses the glycine buffer ultrasonic (under the 200w-5min condition) of pH8.2200mM resuspended after centrifugal 20 minutes, makes latex solution 1.
(B) digestion of IgG sensitizing latex
1) papain of adding 2KU/L in latex solution 1 was 42 ℃ of effects 2 hours;
2) add the papain of 2KU/L, digestive juice 1 is made in continuation effect 2 hours.
(C) Fc fragment sensitizing latex and Fab fragment separates
1) with digestive juice 1 under 4 ℃, 10000g condition centrifugal 20 minutes, abandoning supernatant;
2) the PBS damping fluid 10ml with 50mM cleans twice;
3) resuspended with the PBS damping fluid 5ml ultrasonic (under the 200w-5min condition) of 50mM, make Fc fragment sensitizing latex liquid 2.
(D) Fc fragment sensitizing latex liquid 2 is made RF detectable of the present invention in 60 ℃ of effect hot polymerization sex change in 30 minutes down.
Preparation example 2～8
According to preparation example 1 identical operations, according to changing raw material and condition shown in the table 1, prepare RF detectable of the present invention.
Comparative Examples 1 (preparation reference substance 2):
(A) coupling of the Fc fragment of Calbiochem company and latex particle prepares the RF detectable
1) takes by weighing the Fc fragment of 10.6mg Calbiochem company,, make solution 1 with the PBS damping fluid 5ml dissolving of 100mM;
2) get 10% the polymethylmethacrylate latex of 0.1g particle diameter 135nm, fixed molten with the PBS damping fluid of 50mM to 5ml, make solution 2;
3) solution 2 is slowly joined in the solution 1,40 ℃ of bath temperatures slowly stirred (rotating speed 100rpm) 2 hours;
4) 10000g after the PBS damping fluid 10ml cleaning with 50mM, uses the glycine buffer ultrasonic (under the 200w-5min condition) of pH8.2200mM resuspended after centrifugal 20 minutes, makes Fc sensitizing latex liquid 1.
5) the Fc fragment sensitizing latex is made the RF detectable in 60 ℃ of effect hot polymerization sex change in 30 minutes down.
[experimental example purity and specificity checking]
The checking of experimental example 1 purity
Use the latex solution 2 that obtains in the preparation example 1, the Fc fragment is separated with microballoon, carry out the electrophoretic analysis purity of protein then.
Concrete operations: the latex solution 2 that obtains in the preparation example 1 is centrifugal, abandon supernatant, latex particle draws the PBS damping fluid of the 5mM pH8.0 that leads to X100 (Triton-X100) ultrasonic resuspended with containing 0.1% song, 2-8 ℃ leave standstill 2 hours after, centrifugal 60 minutes of 20000g, obtain supernatant, as Fc fragment of the present invention.Reference substance 1 centrifugal gained supernatant is product Fc fragment in contrast.Adopt Native-PAGE (native polyacrylamide gel electrophoresis) that the gained supernatant is carried out electrophoretic separation, examine with colloid and dye demonstration gel electrophoresis separating resulting, with ImageMaster 2D software analysis collection of illustrative plates.
Analyze the Fc fragment of the present invention of same protein content and the electrophoresis purity of reference substance Fc fragment sample with Native-PAGE, examine the separating resulting that dyes demonstration with colloid and see Fig. 2.
As can be seen from Figure 2, master tape does not have significant difference in the Native-PAGE collection of illustrative plates of Fc fragment of the present invention and two kinds of samples of reference substance Fc fragment, and molecular weight to about the 60KD, is the Fc fragment at 50KD.Based on above-mentioned separating resulting, use the purity of protein (colloid examine dye) of two kinds of samples of ImageMaster 2D software analysis at Native-PAGE, the purity of protein that calculates two kinds of samples under Rolling Disk mode is respectively 97.63% (Fc fragment of the present invention) and 95.67% (reference substance Fc fragment), and purity of the present invention is better than reference substance.
The checking of experimental example 2～5 purity
Use the RF detectable of preparation example 2-5, according to experimental example 1 identical operations, the example 2～5 that experimentizes, the purity of Fc fragment is respectively 97.33%, 96.87%, 96.59%, 98.36% in the RF detectable of the present invention of mensuration.See Table 3.
By above-mentioned experimental example 1～5 as can be seen, the purity height of the Fc fragment by the inventive method preparation is better than the reference substance of prior art, and the separating effect excellence of Fab fragment and Fc fragment in the method for the present invention is described.
The experiment of experimental example 6 specificitys
Experiment reagent:
Use the RF detectable (the 2nd detectable in the table 2) of gained in 50mM PBS damping fluid (the 1st detectable in the table 2) and the preparation example 1 of the present invention to be made into the detectable of the present invention (hereinafter to be referred as " autogamy reagent ") that is used for this experiment;
Use the reference substance 2 (the 2nd detectable in the table 2) of 50mM PBS damping fluid and Comparative Examples 1 preparation gained to be made into the detectable (hereinafter to be referred as " reference substance 2 ") that is used for this experiment, this reference substance is identical with detection principle of the present invention.
The RF detection kit of reference substance 3:Randox (detecting principle is with anti-human rheumatoid factor antibody and RF immune response to take place to detect for RHEUMATOID FACTOR Reagent kit (trade name), immunoturbidimetry RF detection kit).
Detecting instrument: (Olympus AU400 automatic clinical chemistry analyzer).
Detected parameters: the detected parameters of autogamy reagent and reference substance 2 sees Table 2, and the detected parameters of the RF detection kit of Randox is consistent with its product description.
Correlation detection sample: patients serum's sample and 100 routine common patient serum samples that 100 examples are suffered from rheumatoid arthritis;
Accuracy and recovery experiment sample: (RF quality-control product 1:Randox 116LPC (trade name); RF quality-control product 2:Randox 124LPC (trade name); RF quality-control product 3:Randox 134LPC (trade name)), above-mentioned RF quality-control product composition: albumin, α-1 acidoglycoprotein, α-1 antitrypsin, α-2 macroglobulin, alpha-fetoprotein, antistreptolysin O (ASO), Antithrombin III, beta-2 microglobulin, c reactive protein, CER, complement C3, complement C4, ferritin, haptoglobin, immunoglobulin A, immunoglobulin E, immunoglobulin G, immunoglobulin M, the immunoglobulin kappa light chain, the immunoglobulin (Ig) lambda light chain, prealbumin, total protein, RBP ELISA, rheumatoid factor, transferrins.
RF standard items: (RF calibration solution, Randox 1086RF-1090RF (ProductName));
The detected parameters of table 2 autogamy reagent on Olympus AU400 automatic clinical chemistry analyzer
| Reagent | Autogamy RF reagent |
| Sample size (μ l) | ??3 |
| The 1st detectable (μ l) | ??150 |
| The 2nd detectable (μ l) | ??50 |
| Detect wavelength (nm) | ??570 |
| Read a little 1 | ??12 |
| Read a little 2 | ??27 |
Below carry out the specificity of the accuracy and recovery experiment with checking reagent of the present invention.
The accuracy experiment: use autogamy reagent, reference substance 2 and reference substance 3 respectively accuracy and recovery experiment sample to be detected, detect the content of rheumatoid factor, what obtain detection accuracy the results are shown in Table 3.
Table 3
By table 3 result as can be seen, the accuracy in detection result of reagent of the present invention and target value deviation are better than reference substance 2 and reference substance 3 less than 5%.
Recovery experiment: use autogamy reagent, reference substance 2 and reference substance 3 to have the RF standard items (volume ratio of standard items 1087RF and quality-control product 3 is 1: 1) of quality-control product 3 to detect respectively for RF standard items and adding, detect the content of rheumatoid factor, calculate the recovery that RF detects by the measured value that records, the results are shown in Table 4.
Table 4
| Standard items 1087RF initial value C 0(U/ml) | Add quality- |
The recovery (%) | |
| Autogamy reagent | ??15 | ??30.48 | ??100.57 |
| |
??15.1 | ??33.02 | ??111.47 |
| |
??15 | ??31.02 | ??102.93 |
As can be seen from Table 4: the recovery of autogamy reagent reaches 100.57%, illustrate and there is not non-specific disturbance reponse in the detection of RF except other 25 kinds of albumen of rheumatoid factor in the quality-control product that adds, promptly this reagent is better to the specificity of RF, its performance surpasses reference substance 2, reaches Randox reagent level.
Correlativity experiment: in addition, among the present invention, use autogamy reagent, reference substance 2 and reference substance 3 to detect 100 examples and suffer from patients serum's sample of rheumatoid arthritis and the clinical correlation that 100 routine common patient serum samples calculate both, carry out correlation test.The results are shown in Figure 3 and Fig. 4.
According among Fig. 3 and Fig. 4 by the correlativity equation and the relative coefficient that calculate gained, the clinical correlation (R of autogamy reagent and reference substance 3 (Randox detectable)
2=0.9983) is better than the clinical correlation (R of reference substance 2 and reference substance 3
2=0.9885), the specificity for preparing the RF detectable of gained with the inventive method as can be known is better than the specificity of reference substance 2.
By the result of experimental example 6 as can be known, the specificity of the RF detectable by method of the present invention preparation is higher than the specificity that the prior art of same detection principles can reach, i.e. the present invention has not only simplified the preparation method of RF detectable but also improved the specificity that reagent detects RF.
Experimental example 7～10
Use the product of preparation example 5-8, according to experiment Comparative Examples 6 identical operations, the example 7～10 that experimentizes is measured accuracy, the recovery and the clinical correlation of reagent of the present invention, the results are shown in Table 5.
Table 5
Claims (10)
1. the preparation method of rheumatoid factor detection reagent may further comprise the steps:
With IgG molecule and latex particle coupling, obtain the IgG sensitizing latex;
The IgG sensitizing latex that obtains is decomposed with enzymic digestion, obtain the potpourri of Fab fragment and Fc fragment sensitizing latex;
The Fab fragment is separated with the Fc fragment sensitizing latex, obtain the Fc fragment sensitizing latex;
Make Fc fragment sex change in the Fc fragment sensitizing latex, obtain the Fc fragment sensitizing latex of sex change, make rheumatoid factor detection reagent.
2. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, the step of IgG molecule and latex particle coupling is carried out under stirring condition in damping fluid, the buffering range of described damping fluid is at pH6.0-8.5, buffer concentration is 20-100mM, and the stirring revolution is 60-200rpm.
3. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, the step of IgG molecule and latex particle coupling is carried out under 10-40 ℃ temperature.
4. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, described latex particle is the physisorption microballoon, and the particle diameter of latex particle is in the scope of 100-200nm.
5. according to the preparation method of the rheumatoid factor detection reagent of claim 4 record, wherein, the particle diameter of described latex particle is in the scope of 110-180nm.
6. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, the purity of IgG molecule is more than 95%.
7. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, the digestion temperature of IgG molecule sensitizing latex is 30-40 ℃.
8. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, the digestion time of IgG molecule sensitizing latex is 4-5 hour.
9. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, described enzyme is a papain.
10. according to the preparation method of the rheumatoid factor detection reagent of claim 1 record, wherein, separate with the Fc fragment sensitizing latex by the centrifugal Fab of making fragment, centrifugal acceleration is 10000-20000g.
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| CN102901810A (en) * | 2012-10-09 | 2013-01-30 | 北京九强生物技术股份有限公司 | Preparation method of latex particles coated with prostate specific antigen-antibody and PSA enhanced turbidimetric immunophelometry kit |
| CN102901810B (en) * | 2012-10-09 | 2014-08-27 | 北京九强生物技术股份有限公司 | Preparation method of latex particles coated with prostate specific antigen-antibody and PSA enhanced turbidimetric immunophelometry kit |
| CN105001337A (en) * | 2015-07-22 | 2015-10-28 | 丹娜（天津）生物科技有限公司 | Rheumatoid factor adsorbent |
| CN105181962A (en) * | 2015-09-02 | 2015-12-23 | 郁东 | Rheumatoid factor detection reagent |
| CN105181962B (en) * | 2015-09-02 | 2016-09-14 | 郁东 | A kind of rheumatoid factor detection reagent |
| CN109085333A (en) * | 2018-08-22 | 2018-12-25 | 上海复星长征医学科学有限公司 | A kind of preparation, detection kit and the preparation method of rheumatoid factor antigen |
| CN109100515A (en) * | 2018-09-14 | 2018-12-28 | 苏州普瑞斯生物科技有限公司 | A kind of kit and preparation method thereof measuring cyclic citrullinated peptid concentration |
| CN117214428A (en) * | 2023-11-07 | 2023-12-12 | 宁波美康盛德生物科技有限公司 | Rheumatoid factor detection kit and detection method |
| CN117214428B (en) * | 2023-11-07 | 2024-02-02 | 宁波美康盛德生物科技有限公司 | Rheumatoid factor detection kit and detection method |
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