CN102430340B - Method for testing integrity of ultra-filtration membrane envelope - Google Patents
Method for testing integrity of ultra-filtration membrane envelope Download PDFInfo
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- CN102430340B CN102430340B CN 201110360538 CN201110360538A CN102430340B CN 102430340 B CN102430340 B CN 102430340B CN 201110360538 CN201110360538 CN 201110360538 CN 201110360538 A CN201110360538 A CN 201110360538A CN 102430340 B CN102430340 B CN 102430340B
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- milipore filter
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- 238000012360 testing method Methods 0.000 title abstract description 25
- 238000000034 method Methods 0.000 title abstract description 17
- 238000000108 ultra-filtration Methods 0.000 title abstract description 6
- 210000004779 membrane envelope Anatomy 0.000 title abstract 7
- 210000001161 mammalian embryo Anatomy 0.000 claims abstract description 26
- 241000287828 Gallus gallus Species 0.000 claims abstract description 20
- 238000001514 detection method Methods 0.000 claims abstract description 19
- 239000012530 fluid Substances 0.000 claims abstract description 10
- 230000000149 penetrating effect Effects 0.000 claims abstract description 7
- 239000000427 antigen Substances 0.000 claims description 28
- 102000036639 antigens Human genes 0.000 claims description 28
- 108091007433 antigens Proteins 0.000 claims description 28
- 241000711404 Avian avulavirus 1 Species 0.000 claims description 16
- 230000023555 blood coagulation Effects 0.000 claims description 16
- 239000007788 liquid Substances 0.000 claims description 16
- 210000002257 embryonic structure Anatomy 0.000 claims description 12
- 210000001643 allantois Anatomy 0.000 claims description 6
- 238000004321 preservation Methods 0.000 claims description 6
- 238000010561 standard procedure Methods 0.000 claims description 6
- 241000712461 unidentified influenza virus Species 0.000 claims description 5
- 241000700605 Viruses Species 0.000 abstract description 11
- 230000035931 haemagglutination Effects 0.000 abstract description 9
- 239000012528 membrane Substances 0.000 description 12
- 210000004379 membrane Anatomy 0.000 description 11
- 238000000926 separation method Methods 0.000 description 4
- 210000004381 amniotic fluid Anatomy 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000000835 fiber Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 206010011732 Cyst Diseases 0.000 description 2
- 230000004520 agglutination Effects 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 208000031513 cyst Diseases 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000008676 import Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000001471 micro-filtration Methods 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 102000005348 Neuraminidase Human genes 0.000 description 1
- 108010006232 Neuraminidase Proteins 0.000 description 1
- 241000711504 Paramyxoviridae Species 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000012459 cleaning agent Substances 0.000 description 1
- 238000009295 crossflow filtration Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 230000001524 infective effect Effects 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000001728 nano-filtration Methods 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 238000011022 operating instruction Methods 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229940043263 traditional drug Drugs 0.000 description 1
- 210000002845 virion Anatomy 0.000 description 1
Abstract
The invention discloses a method for detecting the integrity of an ultra-filtration membrane envelope; the method comprises the following steps of: firstly, correctly installing a new membrane envelope according to a standard operation method, staring a machine, carrying out HA (hemagglutination) test on withdrawn penetrating fluid after the machine stably operates, if HA is larger than 3log<2>, indicating that the ultra-filtration membrane envelope is incomplete, ending the test; if HA is less than or equal to 3log<2>, and continuously testing; by selecting sensitive virus, inoculating chicken embryo specific to the effluent dialysate of the virus, cultivating the chicken embryo, then determining whether the virus exists in effluent dialysate by a biological detection method, and illustrating whether the ultra-filtration membrane envelope is complete. The method is simple and practical, can carry out on line test, is more accurate in comparison with factory methods and can be used for testing the single membrane envelope, multiple membrane envelopes and systems.
Description
Technical field
The present invention relates to a kind of integrality detection method of film bag, especially a kind of integrality detection method of milipore filter film bag.
Background technology
Membrane separation technique is according to drug activity and its molecular structure and the closely-related characteristic of molecular weight level, by the membrane module of selecting difference to hold back characteristic consist of film separation system to active ingredient separate, concentrate, purifying.Operation can be finished at normal temperatures usually take pressure as motive force, need not to add other material, has advantages of high selectivity.
Film separates and comprises micro-filtration, and ultrafiltration and nanofiltration all are the processes take pressure difference as motive force, and its separating mechanism is that particulate sieves in the fenestra convection cell.Microfiltration membranes be (MF) finger-hole footpath between 0.1-10um, between milipore filter (UF) aperture 0.01-0.1um, NF membrane (NF) aperture is generally between 0.001-0.05um.Usually the particle diameter of virus is generally between 0.01-0.08um, and bacterium liquid, virus have less relative molecular weight, therefore uses the milipore filter concentrated antigen, and is effective, and stability is strong, is suitable for the production and application of certain scale.
The milipore filter screening process, take the pressure differential of film both sides as driving force, take milipore filter as filter medium, under certain pressure, when stoste flow through the film surface, the many tiny micropore that gathers in the milipore filter surface only allowed water and small-molecule substance by becoming through liquid, and volume then is trapped within the liquid feeding side of film in the stoste greater than the material in film surface micropore footpath, become concentrate, thereby realize purification, separation and concentrated purpose to stoste.
When using milipore filter film bag that antigen is concentrated, complete film bag can permeate away hydrone, and enrichment antigen particulate reaches concentrated purpose.
The advantages such as the milipore filter isolation technics has that product yield is high, efficient is high, energy consumption is low, simple to operate, environmental friendliness, in traditional drug production, present significant technical advantage, but because the special macromolecular structure of film regenerates, cleans, utilizes frequency higher, the film Seepage can appear.In fact, because it is complicated to consist of the membrane module installation pipeline of film separation system, tie point is more, therefore is when appearance is unusual in the film course of work, to the leakage detection Comparision difficulty of piece-rate system.
The integrality of film bag is the prerequisite that guarantees separating effect, and the integrity detection of film bag fiber mostly depends on the detection method of film bag producer, and finds in the practical application that producer's detection method detects qualified film bag, and is still bad to viral separating effect.In fact be used for virus and separate, to having relatively high expectations of film bag, the breakage that membrane fiber is trickle can cause virus to reveal, thereby causes antigen losses and to the pollution of environment.
Film integrality detects at present, and the method that generally provides according to equipment producer is carried out the bubble point test method and detected.Open the source of the gas of integrity test device, system is pressurizeed.In system's pressure process, the air velocity that shows on the integrity test device at first will improve, and then drop to a stable flow velocity.The stable flow velocity of this that measures is exactly air forward streams (diffuse flow), if this stable flow velocity has been reduced to and can have accepted under the standard, and consistent with previous value, the film integrality test has been passed through in explanation so.
The concrete detection method that equipment producer provides is as follows:
Before membrane stack was carried out integrity detection, system should be through cleaning and flushing fully, and residual cleaning agent can have a great impact the result.Determine that system is cleaned, after film had drenched fully, intrasystem water is emptying, and the source of the gas that pressure is adjustable was connected on import or the refluxing opening of membrane stack, will not connect import or the refluxing opening sealing of source of the gas, and it is open seeing through the liquid mouth.Slowly be pressurized to appointment air pressure, then stablize and allowed residual water discharge in 5 minutes.Measure and record air pressure, temperature and from seeing through mouth gas flow out, the agent of gas flow available gas flow is measured, the flow and the set quota that relatively detect, if milipore filter is imperfect, its measured air mass flow will substantially exceed the upper limit of velocity parameters in the table.
The report that also has noise measuring method, but because pressure and the flow velocity of tangential flow filtration are larger, the method for noise measuring is inadvisable.
Summary of the invention
Technical problem to be solved by this invention is, a kind of online test method is provided, and can detect the whether integrality detection method of damaged milipore filter film bag of film bag.
In order to solve the problems of the technologies described above, the technical solution used in the present invention is: a kind of integrality detection method of milipore filter film bag may further comprise the steps:
(1) at first new film bag is correctly installed according to the manufacturer's standard method of operating, opened machine, after running is stable, detect;
When (2) detecting, to contain the antigen liquid of the antigen that separates with needs as the NDV that indicates antigen through milipore filter film bag, get film bag diffusate, the penetrating fluid that takes out is HA detects (hemagglutination test (HA test)), survey blood clotting valency HA, if HA＞3log2 illustrates that milipore filter film bag is imperfect, detect and finish; If HA≤3log2 proceeds following step (3) and detects;
(3) get ten ages in days without 10 of the chicken embryos of specific antibodies, wherein inoculate film bag diffusate 0.2ml in 8 every embryo allantois of embryo, stay 2 pieces to compare, inoculating rear 37 ℃ cultivated 120 hours, per sunshine, the chicken embryo was observed 2 times, if the non-specific death of embryo is no more than 1, find that during this time dead embryo places 2-8 ℃ of preservation at once, carry out following step (4) after 120 hours and detect;
(4) all chicken embryos are got blastochyle and measure blood clotting valency HA, if HA≤3log2 illustrates the complete of milipore filter film bag, if HA＞3log2 illustrates the imperfect of milipore filter film bag.
The NDV antigen liquid is counted by volume and is accounted for 0.5-1% in the described antigen liquid.
The described antigen that needs to separate is avian influenza virus or newcastle disease virus.
The invention has the beneficial effects as follows: utilize the sensitiveness of virus, and virus is to erythrocytic agglutination, by detecting agglutination titer, judge the integrality of film bag, method is simple, and the method for comparing producer is more accurate, can be used for the detection of single film bag, the detection of a plurality of film bags and system.With the bioactivity membrane integrity of virus, detection method is simple and easy to do, and need not stop to produce.
The specific embodiment
Below in conjunction with the specific embodiment the present invention is described in further detail:
The integrality detection method of milipore filter film bag of the present invention may further comprise the steps:
(1) at first new film bag is correctly installed according to the manufacturer's standard method of operating, opened machine, after running is stable, detect;
When (2) detecting, to contain the antigen liquid of the antigen that separates with needs as the NDV that indicates antigen through milipore filter film bag, get film bag diffusate, the penetrating fluid that takes out is HA detects (hemagglutination test (HA test)), survey blood clotting valency HA, if HA＞3log2 illustrates that milipore filter film bag is imperfect, detect and finish; If HA≤3log2 proceeds following step (3) and detects;
(3) get ten ages in days without 10 of the chicken embryos of specific antibodies, wherein inoculate film bag diffusate 0.2ml in 8 every embryo allantois of embryo, stay 2 pieces to compare, inoculating rear 37 ℃ cultivated 120 hours, per sunshine, the chicken embryo was observed 2 times, if the non-specific death of embryo is no more than 1, find that during this time dead embryo places 2-8 ℃ of preservation at once, carry out following step (4) after 120 hours and detect;
(4) all chicken embryos are got blastochyle and measure blood clotting valency HA, if HA≤3log2 illustrates the complete of milipore filter film bag, if HA＞3log2 illustrates the imperfect of milipore filter film bag.
The NDV antigen liquid is counted by volume and is accounted for 0.5-1% in the described antigen liquid.
The described antigen that needs to separate is avian influenza virus or newcastle disease virus.
Newcastle disease virus virus (NDV) belongs to Paramyxoviridae, and paramyxovirus genus, nucleic acid are single stranded RNA.Ripe virion is spherical in shape, and diameter is 120-300nm.By the spirality symmetrical disk around nucleocapsid and cyst membrane form.The cyst membrane surface has the fibre of radial arrangement prominent, wherein contains neuraminidase, and therefore virus can be adsorbed on the red blood cell surface of bird, causes erythrocytic aggegation.The median infective dose EID of NDV
50: every 0.1ml viral level 〉=10
8EID
50The particle diameter of newcastle disease virus is less, according to blood coagulation is arranged, still has sensitive blood coagulation phenomenon during the higher titre of dilution, because detection method is simple, can be used as sensitive indicator simultaneously.
And during to the antigen assay with by hemagglutination test (HA test), it is main detecting that HA tires.And measure easy, quick, easy row, be convenient to aborning carry out.
Lower mask body describes:
Measure the method for blood clotting valency HA, claim again the hemagglutination test (HA test) method, see " People's Republic of China's regulations " version hemagglutination test (HA test) in 2000 for details.
The film bag that adopts is purchased from Solution, the molecular cut off 100kd of film bag, and this ultrafiltration membrane stack, molecular cut off 100,000 molecular weight, effective filtration area are 0.5m
2, pressure 0.6MPa.
In addition, can correctly install according to standard operating instructions new film bag, open machine, after running is stable, take off the stream diffusate, survey respectively blood clotting valency HA, as the reference point of integrality.Percentage among the following embodiment is percent by volume.
Embodiment 1
(1) at first new film bag is correctly installed according to the manufacturer's standard method of operating, opened machine, after running is stable, detect;
When (2) detecting, the antigen liquid that will contain 0.5% avian influenza virus that separates with 99.5% needs as the NDV that indicates antigen passes through milipore filter film bag, get film bag diffusate 10ml, the penetrating fluid that takes out is HA detects (hemagglutination test (HA test)), survey blood clotting valency HA, if HA＞3log2 illustrates that milipore filter film bag is imperfect, detect and finish; If HA≤3log2 proceeds following step (3) and detects;
(3) get ten ages in days without 10 of the chicken embryos of specific antibodies, wherein inoculate film bag diffusate 0.2ml in 8 every embryo allantois of embryo, stay 2 pieces to compare, inoculating rear 37 ℃ cultivated 120 hours, per sunshine, the chicken embryo was observed 2 times, if the non-specific death of embryo is no more than 1, find that during this time dead embryo places 2-8 ℃ of preservation at once, carry out following step (4) after 120 hours and detect;
(4) all chicken embryos are got blastochyle (allantoic fluid and amniotic fluid) and measure blood clotting valency HA, if HA≤3log2 illustrates the complete of milipore filter film bag, if HA＞3log2 illustrates the imperfect of milipore filter film bag.
Embodiment 2
(1) at first new film bag is correctly installed according to the manufacturer's standard method of operating, opened machine, after running is stable, detect;
When (2) detecting, the antigen liquid that will contain 1% avian influenza virus that separates with 99% needs as the NDV that indicates antigen passes through milipore filter film bag, get film bag diffusate 10ml, the penetrating fluid that takes out is HA detects (hemagglutination test (HA test)), survey blood clotting valency HA, if HA＞3log2 illustrates that milipore filter film bag is imperfect, detect and finish; If HA≤3log2 proceeds following step (3) and detects;
(3) get ten ages in days without 10 of the chicken embryos of specific antibodies, wherein inoculate film bag diffusate 0.2ml in 8 every embryo allantois of embryo, stay 2 pieces to compare, inoculating rear 37 ℃ cultivated 120 hours, per sunshine, the chicken embryo was observed 2 times, if the non-specific death of embryo is no more than 1, find that during this time dead embryo places 2-8 ℃ of preservation at once, carry out following step (4) after 120 hours and detect;
(4) all chicken embryos are got blastochyle (allantoic fluid and amniotic fluid) and measure blood clotting valency HA, if HA≤3log2 illustrates the complete of milipore filter film bag, if HA＞3log2 illustrates the imperfect of milipore filter film bag.
Embodiment 3
(1) at first new film bag is correctly installed according to the manufacturer's standard method of operating, opened machine, after running is stable, detect;
When (2) detecting, with the antigen liquid process milipore filter film bag of NDV, get film bag diffusate 10ml, the penetrating fluid that takes out is HA detects (hemagglutination test (HA test)), survey blood clotting valency HA, if HA＞3log2, illustrate that milipore filter film bag is imperfect, detect and finish; If HA≤3log2 proceeds following step (3) and detects;
(3) get ten ages in days without 10 of the chicken embryos of specific antibodies, wherein inoculate film bag diffusate 0.2ml in 8 every embryo allantois of embryo, stay 2 pieces to compare, inoculating rear 37 ℃ cultivated 120 hours, per sunshine, the chicken embryo was observed 2 times, if the non-specific death of embryo is no more than 1, find that during this time dead embryo places 2-8 ℃ of preservation at once, carry out following step (4) after 120 hours and detect;
(4) all chicken embryos are got blastochyle (allantoic fluid and amniotic fluid) and measure blood clotting valency HA, if HA≤3log2 illustrates the complete of milipore filter film bag, if HA＞3log2 illustrates the imperfect of milipore filter film bag.
In sum, content of the present invention is not limited in the above-described embodiment, and the knowledgeable people in the same area can propose other embodiment easily within technological guidance's thought of the present invention, but this embodiment comprises within the scope of the present invention.
Claims (2)
1. the integrality detection method of a milipore filter film bag is characterized in that, may further comprise the steps:
(1) at first new film bag is correctly installed according to the manufacturer's standard method of operating, opened machine, after running is stable, detect;
When (2) detecting, will contain the antigen liquid of the antigen that separates with needs as the NDV that indicates antigen through milipore filter film bag, get film bag diffusate, the penetrating fluid that takes out is HA detects, survey blood clotting valency HA, if HA＞3log2, illustrate that milipore filter film bag is imperfect, detect and finish; If HA≤3log2 proceeds following step (3) and detects, the NDV antigen liquid is counted by volume and is accounted for 0.5-1% in the described antigen liquid;
(3) get ten ages in days without 10 of the chicken embryos of specific antibodies, wherein inoculate film bag diffusate 0.2ml in 8 every embryo allantois of embryo, stay 2 pieces to compare, inoculating rear 37 ℃ cultivated 120 hours, per sunshine, the chicken embryo was observed 2 times, if the non-specific death of embryo is no more than 1, find that during this time dead embryo places 2-8 ℃ of preservation at once, carry out following step (4) after 120 hours and detect;
(4) all chicken embryos are got blastochyle and measure blood clotting valency HA, if HA≤3log2 illustrates the complete of milipore filter film bag, if HA＞3log2 illustrates the imperfect of milipore filter film bag.
2. the integrality detection method of milipore filter film bag according to claim 1 is characterized in that, the described antigen that needs to separate is avian influenza virus or newcastle disease virus.
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| CN 201110360538 CN102430340B (en) | 2011-11-15 | 2011-11-15 | Method for testing integrity of ultra-filtration membrane envelope |
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| CN 201110360538 CN102430340B (en) | 2011-11-15 | 2011-11-15 | Method for testing integrity of ultra-filtration membrane envelope |
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| CA3011514C (en) | 2016-01-22 | 2020-11-24 | Baxter International Inc. | Method and machine for producing sterile solution product bags |
| GB2562959A (en) | 2016-01-22 | 2018-11-28 | Baxter Int | Sterile solutions product bag |
| CN108507924B (en) * | 2018-03-30 | 2023-08-04 | 华中科技大学 | Ultrafiltration membrane integrity recognition method and device |
| CN112386949B (en) * | 2020-09-18 | 2022-05-20 | 派特（北京）科技有限公司 | Clinical single-tube fluorine-18 multifunctional module equipment and radiopharmaceutical synthesis process |
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| US5480554A (en) * | 1992-05-13 | 1996-01-02 | Pall Corporation | Integrity-testable wet-dry-reversible ultrafiltration membranes and method for testing same |
| CN2779386Y (en) * | 2005-04-08 | 2006-05-10 | 天津市兴源环境技术工程有限公司 | Membrane module integrity detector for membrane bioreactor |
| FR2894843B1 (en) * | 2005-12-20 | 2008-02-22 | Degremont Sa | METHOD AND APPARATUS FOR INTEGRITY TESTING OF FILTRATION MEMBRANES |
| CN101762561B (en) * | 2009-11-10 | 2012-06-06 | 浙江天元生物药业有限公司 | Method for on-line detecting integrity of ultrafiltration membrane by using ultraviolet monitoring method |
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