CN110157767A - A kind of transfer factor oral solution biological activity determination method - Google Patents
A kind of transfer factor oral solution biological activity determination method Download PDFInfo
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- CN110157767A CN110157767A CN201910314144.7A CN201910314144A CN110157767A CN 110157767 A CN110157767 A CN 110157767A CN 201910314144 A CN201910314144 A CN 201910314144A CN 110157767 A CN110157767 A CN 110157767A
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- liquid
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
Abstract
The present invention provides a kind of transfer factor oral solution biological activity determination method, transfer factor oral solution bioactivity is measured using micro plate-leukocytes adherence inhibition method, calcic magnesium ion in buffer, after drug effect leucocyte, leucocyte relies on the Adherence inhibition factor of calcium ions and magnesium ions by release to inhibit sticking for leucocyte, transfer factor oral solution bioactivity is evaluated by the size of Adhesion inhibiyive effect, verifying of this method Jing Guo scientific system, can be realized the detection to transfer factor oral solution bioactivity.The present invention reflects the immunocompetence of product by the difference of absorption value under measurement leukocyte suspension characteristic absorption wavelength using the high-throughput quickly measurement of 96 hole micro plate combination microplate reader indirectly；Substitute cervical arthroplasty with the measurement result of precision instrument as a result, and the method developed of the present invention do not need that antigen is added, method is simple and easy, and testing cost is low.
Description
Technical field
The invention belongs to multicomponent Biochemical Drugs quality research fields, are related to a kind of transfer factor oral solution bioactivity survey
Determine method.
Background technique
Transfer factor is a kind of cell immunomodulator, is generated by the lymphocyte with cellular immunity function, as
A kind of specific cell factor participates in the immune response of body, and unsensitized lymphocyte transformation can be made with immunocompetent
Lymphocyte, and the function of macrophage can be enhanced, animal body cellular immune function can be improved, promotes body anti-infective and anti-
Imperial ability.The bioactivity of transfer factor is to represent the main indicator of its immunological characteristic.
Conventional method carries out Activity determination using de- E receptor method, but this method is there are more deficiency, as specificity is not strong,
Experiment link complexity, experimental period are compared with long, disturbing factor is more, more demanding to experimenter；Simultaneously because traditional method
Experiment link is more complicated, and the experimental material being related to is relatively more, especially biomaterial leucocyte and red blood cell, and testing result
Judged by the way of microscopy, is taken a part for the whole；Therefore traditional technique in measuring is used, as a result reproducibility is poor.Therefore, it establishes fast
Speed, easy and sensitive activity test method are for evaluating the height of transfer factor efficiency and the control of quality with particularly significant
Meaning.
Summary of the invention
Aiming at the problems existing in the prior art, the present invention provides a kind of transfer factor oral solution biological activity determination side
Method, this method is easy to operate, and favorable reproducibility, specificity is strong, and detection speed is fast.
The present invention is to be achieved through the following technical solutions:
A kind of transfer factor oral solution biological activity determination method, which comprises the following steps:
Step 1, Hank ' s liquid is prepared, leukocyte suspension is prepared；
Step 2, be measured in culture plate: test sample is added in test sample hole, and Hank ' s liquid is added in control wells, then often
Leukocyte suspension is added in hole, and culture plate is placed culture in the incubator and is incubated for, culture takes out culture plate juxtaposition after the completion of being incubated for
In shaking in concussion instrument, nonadherent cell supernatant is shaken and has been sucked out, suspension after being shifted measures every at 405nm
Absorbance value of the absorbance of suspension as cell after adherency after the transfer in hole is assigned to obtain the extinction of cell after test sample adheres to
The absorbance value of cell after angle value and reference substance adherency；Test sample and leukocyte suspension is added in test sample hole before adhering to, before adherency
Hank ' s liquid and leukocyte suspension is added in control wells, and the absorbance that every hole sample is measured at 405nm is precellular as adhering to
Absorbance value respectively obtains test sample and adheres to precellular absorbance value and the precellular absorbance value of reference substance adherency；It utilizes
Formula (1) calculates separately to obtain the adhesion rate of the adhesion rate of test sample and reference substance, recycles formula (2) that test sample is calculated
Adhesion inhibition rate to get arrive test sample bioactivity size；
Preferably, in step 1, leukocyte suspension is prepared using rabbit thymus gland or pig lymph.
Preferably, leukocyte suspension is prepared using rabbit thymus gland.
Preferably, in step 1, leukocyte suspension is prepared method particularly includes: take rabbit thymus gland or pig lymph, shred, add Hank '
Cell suspension is made in s liquid, filters, centrifugation, discards supernatant liquid, and precipitating plus Hank ' s liquid are uniformly dispersed, mix with cell separating liquid,
Centrifugation, is sucked out the thymocyte of middle layer, and thymocyte is washed with Hank ' s liquid, centrifugation, discards supernatant liquid, and precipitating is used again
The dilution of Hank ' s liquid, obtains leukocyte suspension.
Preferably, in step 1, leukocyte suspension concentration is to contain (1 × 10 in every 1mL7)~(4 × 107) a cell.
Preferably, in step 2, leucocyte is added in test sample hole and the preceding control wells of adherency before test sample hole, control wells, adherency
The volume of suspension is 50~100 μ L.
Preferably, in step 2, culture incubation time is 2~4 hours.
Preferably, in step 2, culture plate is placed on 37 degrees Celsius of cultures in 5% carbon dioxide incubator.With the prior art
It compares, the invention has the following beneficial technical effects:
The present invention measures transfer factor oral solution bioactivity, buffer using micro plate-leukocytes adherence inhibition method
Middle calcic magnesium ion, after drug effect leucocyte, leucocyte relies on the Adherence inhibition factor of calcium ions and magnesium ions by release to press down
Leucocyte processed sticks, and evaluates transfer factor oral solution bioactivity by the size of Adhesion inhibiyive effect, this method is passed through
The verifying of scientific system can be realized the detection to transfer factor oral solution bioactivity.The present invention uses 96 hole micro plates
It is reversed by the difference of absorption value under measurement leukocyte suspension characteristic absorption wavelength in conjunction with the high-throughput quickly measurement of microplate reader
Reflect the immunocompetence of product；Substitute cervical arthroplasty with the measurement result of precision instrument as a result, and the method developed of the present invention be not required to
Antigen is added, method is simple and easy, and testing cost is low.Present invention reduces the artificial subjective factor of detection process, it is easy to slap
Hold, convenient for promoting, while it is more accurate compared with de- E receptor method, quickly, favorable reproducibility, specificity is strong, method is simple and easy, detects
As a result accurate, reliable.The present invention is suitable for the detection and monitoring of transfer factor oral solution bioactivity.
Further, by screening the preferably new fresh rabbit thymus gland in lymphocyte source, because fresh porcine thymus is in routine check
In be not easy to obtain, and using pig lymph extract leukocyte power it is poor compared with rabbit thymus gland, blank control adherence rate is low；Therefore sample
The lesser difference of product adherence rate can lead to the vigor result i.e. biggish difference of Adherence inhibition rate, therefore use pig lymph testing result
Poor reproducibility；And use rabbit thymus gland detection control adhesion rate higher, operator is required low, and checkability is higher.
Further, by screening leukocyte suspension concentration, and experimental result is analyzed with statistics, it is determined that
(1 × 10 in the preferably every 1mL of leukocyte suspension ultimate density7)~(4 × 107) a cell, Adhesion inhibiyive effect becomes apparent from, detects
As a result more reliable.
Further, volume is added by screening leukocyte suspension, and experimental result is analyzed with statistics, really
Having determined optimal leukocyte suspension volume is 50~100 μ L, and Adhesion inhibiyive effect becomes apparent from, and testing result is more reliable.
Further, by screening incubation time, and experimental result is analyzed with statistics, it is determined that preferred
Incubation time is 2~4 hours, and Adhesion inhibiyive effect becomes apparent from, and testing result is more reliable.
Specific embodiment
Below with reference to specific embodiment, the present invention is described in further detail, it is described be explanation of the invention and
It is not to limit.
Transfer factor oral Solution Active detection of the present invention uses micro plate-leukocytes adherence inhibition method, including following step
It is rapid:
A, potassium dihydrogen phosphate 0.06g, sodium chloride 8.0g, sodium bicarbonate 0.35g, potassium chloride the preparation of Hank ' s liquid: are weighed
0.4g, glucose sugar 1.0g, disodium hydrogen phosphate 0.14g, calcium chloride 0.14g, magnesium sulfate 0.1g, magnesium chloride 0.1g are dissolved in water into
1000mL, with concentrated sulfuric acid tune pH to 6.5.It is saved 1 month for 4 DEG C after 115 DEG C of 20min sterilizings.It is molten with 5.6% sodium bicarbonate before use
Liquid tune pH to 7.2~7.4.
B, the preparation of lymphocyte suspension: taking new fresh rabbit thymus gland or pig lymph, shred, and appropriate Hank ' s liquid is added to make into cell
Suspension, through 200 mesh filter, 1500r/min be centrifuged 3~5 minutes, discard supernatant liquid, a small amount of Hank ' s liquid added to beat, with account for its 1/
The cell separating liquid of 2 volumes mixes, and is centrifuged 20min with 2000r/min, the careful thymocyte that middle layer is sucked out is put into another
In centrifuge tube, add appropriate Hank ' s liquid to wash, shake up, with 1500r/min centrifugation 3~5 minutes, liquid is discarded supernatant, with Hank ' s
Liquid washs (operation is the same) three times, then is diluted and counted with Hank ' s liquid, makes (1 × 10 in the every 1mL of ultimate density7)~(4 ×
107) a cell, as leukocyte suspension.
C, measuring method: carrying out in 96 well culture plates, and test sample does 12 holes, and reference substance does 4 holes, and the every hole in test sample hole is added
50 μ L of Hank ' s liquid is added in 50 μ L of test sample, control wells, and then 50~100 μ L of leukocyte suspension is added in every hole, and culture plate is put
It sets 37 degrees Celsius of cultures in 5% carbon dioxide incubator to be incubated for 2~4 hours, 50r/min shakes in horizontal concussion instrument after taking-up
1~3min is swung, nonadherent cell has been shaken to and has been sucked out supernatant transfer, every hole adds 50 μ L of Hank ' s liquid, washes by upper method
It washs once, washing lotion is merged with supernatant, suspension after being shifted.Using the high-throughput quickly measurement of microplate reader, surveyed at 405nm
Absorbance value of the absorbance of suspension as cell after adherency, assigns to obtain the suction of cell after test sample adheres to after fixed every hole transfer
The absorbance value of cell after shading value and reference substance adherency.4 holes are respectively done before test sample and reference substance adherency, test sample hole before adhering to
50 μ L of test solution and 50~100 μ L of leukocyte suspension is added, control wells addition takes 50 μ L of Hank ' s liquid and white thin before adhering to
50~100 μ L of born of the same parents' suspension measures the absorbance in every hole as precellular absorbance value is adhered at 405nm, respectively obtains confession
Test product adheres to precellular absorbance value and adheres to precellular absorbance value for reference substance.It is calculated by following formula, utilizes public affairs
Formula (1) calculates separately to obtain the adhesion rate of the adhesion rate of test sample and reference substance, recycles formula (2) that test sample is calculated
Adhesion inhibition rate, the adhesion inhibition rate of test sample are to reflect the bioactivity size of test sample.
In step a, prepares Hank ' s liquid and contain calcium and magnesium ion, the Adhesion inhibiyive effect of such test sample becomes apparent from, i.e. vigor
Value is higher, and testing result is more reliable.
In step b, preferably new fresh rabbit thymus gland, because fresh porcine thymus is not easy to obtain in routine check, using pig lymph
Testing result poor reproducibility；And use rabbit thymus gland detection control adhesion rate higher, operator is required low, and checkability is more
It is high.
In step b, (1 × 10 in the preferably every 1mL of leukocyte suspension ultimate density7)~(4 × 107) a cell, by examining
It examines, 2 × 10 is contained in most preferably every 1mL7A cell, as leukocyte suspension.
In step c, leukocyte suspension volume preferably 50~100 μ L are added, because by investigating, most preferably 100 μ L, detection
As a result more reliable.
In step c, incubation time preferably 2~4 hours, most preferably 3 hours.
The screening for being incubated for leukocyte suspension volume is added in 1 micro plate of embodiment-leukocytes adherence inhibition method determination condition
Step:
A, potassium dihydrogen phosphate 0.06g, sodium chloride 8.0g, sodium bicarbonate 0.35g, potassium chloride the preparation of Hank ' s liquid: are weighed
0.4g, glucose sugar 1.0g, disodium hydrogen phosphate 0.14g, calcium chloride 0.14g, magnesium sulfate 0.1g, magnesium chloride 0.1g are dissolved in water into
1000mL, with concentrated sulfuric acid tune pH to 6.5.It is saved 1 month for 4 DEG C after 115 DEG C of 20min sterilizings.It is molten with 5.6% sodium bicarbonate before use
Liquid tune pH to 7.2.
B, the preparation of lymphocyte suspension: taking new fresh rabbit thymus gland, shred, and appropriate Hank ' s liquid is added to make into cell suspension, warp
The filtering of 200 mesh, 1500r/min are centrifuged 3 minutes, discard supernatant liquid, a small amount of Hank ' s liquid is added to beat, and account for the thin of its 1/2 volume
The mixing of born of the same parents' separating liquid is centrifuged 20min with 2000r/min, and the careful thymocyte that middle layer is sucked out is put into another centrifuge tube,
Add appropriate Hank ' s liquid to wash, shake up, with 1500r/min centrifugation 3 minutes, discard supernatant liquid, washs (behaviour three times with Hank ' s liquid
Make the same), then diluted and counted with Hank ' s liquid, make 2 × 10 in the every 1mL of ultimate density7A cell, it is outstanding as leucocyte
Liquid.
C, measuring method: carrying out in 96 well culture plates, and test sample does 12 holes, and reference substance does 4 holes, and the every hole in test sample hole is added
50 μ L of Hank ' s liquid is added in 50 μ L of test sample, control wells, and then leukocyte suspension is added in every hole, and culture plate is placed on 5% 2
37 degrees Celsius of cultures are incubated for 2 hours in carbonoxide incubator, and 50r/min shakes 1min in horizontal concussion instrument after taking-up, will not
The cell of adherency has shaken and has been sucked out supernatant transfer, and every hole adds 50 μ L of Hank ' s liquid, washed once by upper method, by washing lotion with
Supernatant merges, suspension after being shifted.Using the high-throughput quickly measurement of microplate reader, every hole is measured at 405nm and shifts rear overhang
Absorbance value of the absorbance of liquid as cell after adherency is assigned to obtain the absorbance value and reference substance of cell after test sample adheres to
The absorbance value of cell after adherency.4 holes are respectively done before test sample and reference substance adherency, test solution is added in test sample hole before adhering to
50 μ L and leukocyte suspension, control wells addition takes 50 μ L of Hank ' s liquid and leukocyte suspension before adhering to, and every hole is measured at 405nm
Absorbance as precellular absorbance value is adhered to, respectively obtain test sample and adhere to precellular absorbance value and for reference substance
Adhere to precellular absorbance value.The adhesion inhibition rate of test sample is calculated by formula above.
Through two experimenters on the basis of other experimental implementations are constant, the investigation of same batch transfer factor solution is incubated
Testing result when leukocyte suspension addition volume is respectively 50 μ L and 100 μ L is educated, every experimenter is measured in parallel 6 groups of numbers
According to statistic mixed-state the results are shown in Table 1, wherein 95%CI is 95% confidence interval, and FL is Reliable limit rate.
The investigation result for being incubated for leucocyte volume is added in table 1
From the above experiments, it was found that testing result is more reliable when addition incubation leukocyte suspension volume is 100 μ L, therefore
Most preferably being added and being incubated for leukocyte suspension volume is 100 μ L.
The screening of incubation time in 2 micro plates of embodiment-leukocytes adherence inhibition method measurement bioactivity
Step:
A, potassium dihydrogen phosphate 0.06g, sodium chloride 8.0g, sodium bicarbonate 0.35g, potassium chloride the preparation of Hank ' s liquid: are weighed
0.4g, glucose sugar 1.0g, disodium hydrogen phosphate 0.14g, calcium chloride 0.14g, magnesium sulfate 0.1g, magnesium chloride 0.1g are dissolved in water into
1000mL, with concentrated sulfuric acid tune pH to 6.5.It is saved 1 month for 4 DEG C after 115 DEG C of 20min sterilizings.It is molten with 5.6% sodium bicarbonate before use
Liquid tune pH to 7.2.
B, the preparation of lymphocyte suspension: taking new fresh rabbit thymus gland, shred, and appropriate Hank ' s liquid is added to make into cell suspension, warp
The filtering of 200 mesh, 1500r/min are centrifuged 3 minutes, discard supernatant liquid, a small amount of Hank ' s liquid is added to beat, and account for the thin of its 1/2 volume
The mixing of born of the same parents' separating liquid is centrifuged 20min with 2000r/min, and the careful thymocyte that middle layer is sucked out is put into another centrifuge tube,
Add appropriate Hank ' s liquid to wash, shake up, with 1500r/min centrifugation 3 minutes, discard supernatant liquid, washs (behaviour three times with Hank ' s liquid
Make the same), then diluted and counted with Hank ' s liquid, make 1 × 10 in the every 1mL of ultimate density7A cell, it is outstanding as leucocyte
Liquid.
C, measuring method: carrying out in 96 well culture plates, and test sample does 12 holes, and reference substance does 4 holes, and the every hole in test sample hole is added
50 μ L of Hank ' s liquid is added in 50 μ L of test sample, control wells, and then 100 μ L of leukocyte suspension is added in every hole, and culture plate is placed on
37 degrees Celsius of cultures are incubated in 5% carbon dioxide incubator, and 50r/min shakes 1min in horizontal concussion instrument after taking-up, will not
The cell of adherency has shaken and has been sucked out supernatant transfer, and every hole adds 50 μ L of Hank ' s liquid, washed once by upper method, by washing lotion with
Supernatant merges, suspension after being shifted.Using the high-throughput quickly measurement of microplate reader, every hole is measured at 405nm and shifts rear overhang
Absorbance value of the absorbance of liquid as cell after adherency is assigned to obtain the absorbance value and reference substance of cell after test sample adheres to
The absorbance value of cell after adherency.4 holes are respectively done before test sample and reference substance adherency, test solution is added in test sample hole before adhering to
100 μ L of 50 μ L and leukocyte suspension, control wells addition takes 100 μ L of 50 μ L of Hank ' s liquid and leukocyte suspension before adhering to, in 405nm
Place measures the absorbance in every hole as precellular absorbance value is adhered to, and respectively obtains test sample and adheres to precellular absorbance value
Precellular absorbance value is adhered to for reference substance.The adhesion inhibition rate of test sample is calculated by formula above.
Above-mentioned implementation steps, in the case where other operating parameters are constant, to same batch transfer factor oral solution, respectively
The adhesion inhibition rate of 2h, 3h and 4h totally three different incubation time points is investigated, each incubation time point is measured in parallel 9 groups of numbers
According to statistic mixed-state the results are shown in Table 2, wherein 95%CI is 95% confidence interval, and FL is Reliable limit rate.
The investigation result of 2 incubation time of table
From the above experiments, it was found that incubation time, which is appropriately extended, can be improved the adhesion inhibition rate of sample and can be improved
The reliability of testing result is incubated for the testing result of 3h and 4h without significant difference, therefore to improve detection efficiency, is most preferably incubated for
Time is 3h.
3 micro plates of embodiment-leukocytes adherence inhibition method measurement bioactivity accuracy is investigated
Leukocyte suspension volume is 100 μ L, and leukocyte suspension concentration is 4 × 10 in every 1mL7A cell, other conditions with
Embodiment 1 is identical, will with batch transfer factor oral solution with Hank ' s liquid be diluted to 5 series of concentrations solution (5%,
10%, 50%, 75%, 100%) determination of activity is carried out, the relationship between test sample concentration and activity is investigated, to judge this hair
The accuracy of bright method.Each concentration is measured in parallel at least 6 times, is analyzed with statistics experimental result, statistic mixed-state
It the results are shown in Table 3, wherein 95%CI is 95% confidence interval, and FL is Reliable limit rate.
The investigation result of 3 drug concentration of table and activity relationship
Regression analysis is carried out to said medicine concentration (independent variable) and Activity Results (dependent variable), obtaining regression equation is C2
=0.879+47.46C1, related coefficient is 86.34% after adjustment；And P < 0.05, illustrate that independent variable drug concentration is living to dependent variable
Property result explanation strengths or predictive power be positively correlated, related coefficient adjusted be 0.8634.Illustrate the detection side that the present invention develops
Method has more significant distinction, i.e. testing result is accurately and reliably.
Claims (8)
1. a kind of transfer factor oral solution biological activity determination method, which comprises the following steps:
Step 1, Hank ' s liquid is prepared, leukocyte suspension is prepared；
Step 2, be measured in culture plate: test sample is added in test sample hole, and Hank ' s liquid is added in control wells, and then every hole adds
Enter leukocyte suspension, culture plate is placed into culture in the incubator and is incubated for, culture takes out culture plate after the completion of being incubated for and is placed in shake
It swings and is shaken on instrument, nonadherent cell has been shaken and be sucked out supernatant, suspension after being shifted measures every hole at 405nm
Absorbance value of the absorbance of suspension as cell after adherency after transfer is assigned to obtain the absorbance value of cell after test sample adheres to
With the absorbance value of cell after reference substance adherency；Test sample and leukocyte suspension is added in test sample hole before adhering to, control before adhering to
Hank ' s liquid and leukocyte suspension is added in hole, and the absorbance of every hole sample is measured at 405nm as the precellular extinction of adherency
Angle value respectively obtains test sample and adheres to precellular absorbance value and the precellular absorbance value of reference substance adherency；Utilize formula
(1) it calculates separately to obtain the adhesion rate of test sample and the adhesion rate of reference substance, recycles formula (2) that the viscous of test sample is calculated
Attached inhibiting rate is to get the bioactivity size for arriving test sample；
2. transfer factor oral solution biological activity determination method according to claim 1, which is characterized in that in step 1,
Leukocyte suspension is prepared using rabbit thymus gland or pig lymph.
3. transfer factor oral solution biological activity determination method according to claim 1, which is characterized in that use rabbit chest
Gland prepares leukocyte suspension.
4. transfer factor oral solution biological activity determination method according to claim 1, which is characterized in that in step 1,
Prepare leukocyte suspension method particularly includes: rabbit thymus gland or pig lymph are taken, is shredded, adds Hank ' s liquid that cell suspension is made, is filtered, from
The heart discards supernatant liquid, and precipitating plus Hank ' s liquid are uniformly dispersed, mix with cell separating liquid, are centrifuged, and the thymus gland that middle layer is sucked out is thin
Born of the same parents, thymocyte are washed with Hank ' s liquid, centrifugation, discard supernatant liquid, and precipitating is diluted with Hank ' s liquid again, and it is outstanding to obtain leucocyte
Liquid.
5. transfer factor oral solution biological activity determination method according to claim 1, which is characterized in that in step 1,
Leukocyte suspension concentration is to contain (1 × 10 in every 1mL7)~(4 × 107) a cell.
6. transfer factor oral solution biological activity determination method according to claim 1, which is characterized in that in step 2,
It is 50~100 μ that the volume of leukocyte suspension, which is added, in test sample hole and the preceding control wells of adherency before test sample hole, control wells, adherency
L。
7. transfer factor oral solution biological activity determination method according to claim 1, which is characterized in that in step 2,
Cultivating incubation time is 2~4 hours.
8. transfer factor oral solution biological activity determination method according to claim 1, which is characterized in that in step 2,
Culture plate is placed on 37 degrees Celsius of cultures in 5% carbon dioxide incubator.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0301992A2 (en) * | 1987-07-30 | 1989-02-01 | Centro Nacional De Biopreparados | Vaccine against group B Neisseria meningitidis, gammaglobulin and transfer factor |
-
2019
- 2019-04-18 CN CN201910314144.7A patent/CN110157767A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0301992A2 (en) * | 1987-07-30 | 1989-02-01 | Centro Nacional De Biopreparados | Vaccine against group B Neisseria meningitidis, gammaglobulin and transfer factor |
Non-Patent Citations (3)
| Title |
|---|
| 王灿等: "白细胞黏附抑制实验在核糖核酸药物免疫活力测定中的应用研究", 《中国药师》 * |
| 韩彩霞等: "抗乙肝特异性转移因子和核糖核酸体外生物活性的实验研究 Ⅲ.用白细胞移动抑制试验测活", 《中国生化药物杂志》 * |
| 黄青等: "转移因子脱E受体活力测定方法的优化及基于微量板-白细胞黏附抑制试验的新方法研究", 《中国药学杂志》 * |
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