CN206736270U - Pig thrombiase production system - Google Patents

Abstract

The utility model provides pig thrombiase production system, including blood Disengagement zone, anion exchange area, activation liquid reaction tank, inactivation of virus area, cation exchange area and the virus filtration area sequentially connected；Wherein, activation liquid reaction tank is furnished with two feeding devices with metering valve, and two feeding devices are respectively provided with calcium chloride and lecithin.Compared with prior art, pig thrombiase operation system structure provided by the utility model is simple, new blood, which only needs once to purify, can make fibrin ferment purity reach >=500 units/mg Solid contents, gained thrombin solution can be directly used for the production of fibrin ferment freeze-dried powder, need not be secondarily purified, substantially reduce production cost.

Description

Pig thrombiase production system

Technical field

It the utility model is related to biological technical field, more particularly to the production equipment of pig thrombiase.

Background technology

Fibrin ferment is a kind of serine protein hydrolase with high degree of specificity in blood plasma blood coagulation system, can be used as office Portion's hemostatic, it is the bulk drug of fibrin ferment freeze-dried powder.Fibrin ferment can be made molten with 4 former Arg-Gly peptide bonds of hydrolysis of fibrin The fibrinogen of colloidal state is changed into the fibrin monomer of gel state, promotes blood clotting, while can promote secretion of platelet And aggegation, blood clotting is further speeded up, reaches the purpose stopped blooding rapidly, therefore, fibrin ferment is in blood clotting and hemostasis In play an important role.Clinically, fibrin ferment is widely used in local hemostasis, small available for being not easy to ligature in operation Vascular haemostasis, hemorrhage of digestive tract and traumatism and bleeding etc..

The thrombin preparation clinically used at present is largely the factor extracted from ox blood or pig blood, through blood coagulation Activation of zymogen thing activates and obtains the aseptic freeze-dried product of fibrin ferment, has higher selectivity, is a kind of quick-acting local hemostatics, extensively It is general that applied to hemorrhage of digestive tract and surgical operation hemostasis, (Song Hongxin, Ma Yongzheng, Li Minkang separating and purifying method for thrombase is ground Study carefully progress [J] food research and developments, 2004,25 (6)：65-67.).Prior art preparation method generally comprises three substantially Step, that is, extract, purify and freeze (or preparation).Publication No. CN1844380A patent of invention propose one kind prepare it is high-purity The method of fibrin ferment is spent, this method is eluted to chromatographic column, collected solidifying using chromatography media absorption low-purity thrombin solution Hemase protein peak, then through gel chromatography, desalination and concentration by ultrafiltration, finally freeze to obtain restricted fibrin ferment finished product.The patent application is only Use the purity for than a parameter living, being not enough to illustrate fibrin ferment.

And the process that isolates and purifies of pig thrombiase generally comprises the pretreatment of Swine plasma, the extraction of factor, fibrin ferment The step such as former activation and the preparation of thick fibrin ferment, the purifying of fibrin ferment.Because blood coagulation enzyme preparation is to extract to prepare from pig blood Obtained by biochemical drug, it is potential have pig borne virus pollute hidden danger, for improve products safety, it is necessary to carry out blood coagulation enzymophathy Malicious inactivation technology research；《Chinese Pharmacopoeia》" fibrin ferment freeze-dried powder " quality standard that version two page 1561 in 2015 is recorded【System Method requirement】Item regulation：" this product is animal origin, should effectively remove method and the measure of virus or inactivation of virus ".Publication number A kind of preparation method of fibrin ferment is disclosed for CN104328101A Chinese invention patent, this method uses virus-inactivating agent The mode that inactivation treatment and nanofiltration membrane filtering are combined, to inactivate and remove potential pig borne virus pollution in product, take Obtain beneficial effect necessarily.But this method has that processing step is more, complicated, and PROTHROMBIN ACTIVATOR method is complicated, cumbersome, it is impossible to keeps away Exempt from the shortcomings that exogenous virus secondary pollution risk be present during thrombin preparation.

Further, since the key of fibrin ferment preparation technology is purity, prior art usually requires purifying two to three times, just may be used The fibrin ferment of preparation is reached purity requirement, considerably increase production cost.

Utility model content

The purpose of this utility model is the shortcomings that overcoming above-mentioned prior art, there is provided a kind of once purifying concentration can make pure Degree reaches the production system of the pig thrombiase of >=500 units/mg Solid contents, and obtained pig thrombiase inactivation of virus, reaches National standard.

To achieve these goals, technical scheme provided by the utility model is：

Pig thrombiase production system, including sequentially connect blood Disengagement zone, anion exchange area, activation liquid reaction tank, Inactivation of virus area, cation exchange area and virus filtration area；Wherein, liquid reaction tank is activated equipped with two charging dresses with metering valve Put, two feeding devices are respectively provided with calcium chloride and lecithin, are coagulated in cation exchange area provided with CM glucan cation exchanges Glue.

Preferably, pig thrombiase production system also includes lyophilized area, for the thrombin solution of preparation to be freezed.

Preferably, a scrubbing section is connected between anion exchange area and activation liquid reaction tank, for eluting anion exchange Gel.

Preferably, a scrubbing section is connected between cation exchange area and virus filtration area, is coagulated for eluting cation exchange Glue.

Preferably, blood Disengagement zone includes anti-freezing portion and the blood separation unit of connection, and new blood adds into anti-freezing portion Blood separation unit is entered back into after anti-coagulants, is centrifuged in blood separation unit, it is standby to collect anti-freezing blood plasma.

It is highly preferred that blood separation unit is a blood separator, for new blood to be centrifuged.

Preferably, anion exchange area is a deae dextran anion-exchange gel, anti-freezing blood plasma deae dextran Anion-exchange gel adsorbs factor, and prothrombin solution is obtained after elution.

Preferably, inactivation of virus area includes ultrafiltration concentration portion, removal of impurities portion and the S/D inactivation of virus portion sequentially connected, blood coagulation After proenzyme solution is concentrated by ultrafiltration in ultrafiltration concentration portion, the filter through different pore size in removal of impurities portion carries out filtering and impurity removing, finally Inactivation of virus is carried out into S/D inactivation of virus portion.

It is highly preferred that removal of impurities portion includes spaced at least three filter membrane, the aperture of filter membrane is gradually reduced；It is preferred that filter Membrane aperture is followed successively by 5 μm, 3 μm, 1.2 μm and 0.8 μm.

It is highly preferred that S/D inactivation of virus portion is a S/D process tanks.

Preferably, cation exchange area is a CM glucan cation exchange gels, and the S/D after S/D inactivation of virus coagulates Hemase solution after CM glucan cation exchange gel adsorptions with eluting to obtain purifying blood coagulation enzyme solutions.

Preferably, virus filtration area includes connection ultrafiltration concentration portion and virus filtration portion, and purifying blood coagulation enzyme solutions are in ultrafiltration After concentrating part is concentrated by ultrafiltration, virus filtration is carried out through virus filtration portion.

It is highly preferred that virus filtration portion removes virus filter from 20nm.

Preferably, the design parameter of CM glucans cation exchange gel is：

Outward appearance：White particle, odorless, tasteless, it is visible by naked eyes impurity；

Species：Weak cation exchanger；

Matrix：Sephadex G -50 (seplite G-50)；

Main function functional group：Carboxymethyl；

Form：Sodium form；

Particle size range：40~120 μm (dry glues)；

Applicable pH range：6~10；

Peak flow rate (PFR)：≥45cm/h；

Carrying capacity：>=120mg ribalgilases/mL；

Ion exchange capacity：4.0~5.0mmol/g dry glues.

Compared with prior art, pig thrombiase operation system structure provided by the utility model is simple, and new blood only needs Once purifying can make fibrin ferment purity reach >=500 units/mg Solid contents, and gained thrombin solution can be directly used for fibrin ferment The production of freeze-dried powder, without secondarily purified, substantially reduce production cost.

Brief description of the drawings

Fig. 1 is the pig thrombiase production system topological diagram that the utility model embodiment provides.

Embodiment

In order to more clearly describe technology contents of the present utility model, enter traveling one with reference to specific embodiment The description of step.

Pig thrombiase production system provided by the utility model, including sequentially connect blood Disengagement zone, anion exchange Area, the first scrubbing section, activation liquid reaction tank, inactivation of virus area, cation exchange area, the second scrubbing section and virus filtration area；Swash Liquid reaction tank living is furnished with two feeding devices with metering valve, and calcium chloride and lecithin are respectively provided with two feeding devices.

Wherein, it is a DEAE that blood Disengagement zone, which includes the anti-freezing portion sequentially connected and blood separator, anion exchange area, Dextran anion exchanges gel, and inactivation of virus area includes the first ultrafiltration concentration portion, removal of impurities portion and the S/D processing sequentially connected Tank, removal of impurities portion are followed successively by 5 μm, 3 μm, 1.2 μm and 0.8 μm specification filter membranes, and cation exchange area hands over for a CM glucans cation Gel is changed, virus filtration area includes the second ultrafiltration concentration portion of connection and 20nm removes virus filter.

During the pig thrombiase production system work, new blood enters back into blood after entering anti-freezing portion addition anti-coagulants Separation unit, centrifuged in blood separation unit, collection anti-freezing blood plasma is standby, and anti-freezing blood plasma is coagulated with deae dextran anion exchange Glue adsorbs factor, and prothrombin solution is obtained after elution, after prothrombin solution is concentrated by ultrafiltration in the first ultrafiltration concentration portion, warp Removal of impurities portion carries out filtering and impurity removing built with the filter of different pore size filter membrane, finally enters S/D inactivation of virus portion and carries out virus Inactivation, S/D thrombin solutions after S/D inactivation of virus with eluted after CM glucan cation exchange gel adsorptions must purify it is solidifying Hemase solution, after purifying blood coagulation enzyme solutions are concentrated by ultrafiltration in the second ultrafiltration concentration portion, virus filtration, institute are carried out through virus filtration portion Obtaining thrombin solution, finally freeze-dried area freezes, and obtains fibrin ferment and freezes finished product.

In this description, the utility model is described with reference to its specific embodiment.But it is clear that still can be with Various modification can be adapted and conversion is without departing from spirit and scope of the present utility model.Therefore, specification and drawings are considered as It is illustrative and not restrictive.

Claims (10)

1. pig thrombiase production system, it is characterised in that：Including the blood Disengagement zone, anion exchange area, activation sequentially connected Liquid reaction tank, inactivation of virus area, cation exchange area and virus filtration area；Wherein, liquid reaction tank is activated to measure equipped with two bands The feeding device of valve, two feeding devices are respectively provided with calcium chloride and lecithin, CM glucans sun are provided with cation exchange area Ion-exchange gel.

2. the pig thrombiase production system described in claim 1, it is characterised in that：Also include lyophilized area.

3. the pig thrombiase production system described in claim 1, it is characterised in that：Anion exchange area and activation liquid reaction tank it Between connect a scrubbing section.

4. the pig thrombiase production system described in claim 1, it is characterised in that：Between cation exchange area and virus filtration area Connect a scrubbing section.

5. the pig thrombiase production system described in claim 1, it is characterised in that：Blood Disengagement zone include connection anti-freezing portion and Blood separation unit.

6. the pig thrombiase production system described in claim 1, it is characterised in that：Inactivation of virus area includes the ultrafiltration sequentially connected Concentrating part, removal of impurities portion and S/D inactivation of virus portion.

7. the pig thrombiase production system described in claim 1, it is characterised in that：Removal of impurities portion selects the filter membrane that aperture sequentially reduces Filter successively.

8. the pig thrombiase production system described in claim 1, it is characterised in that：S/D inactivation of virus portion is a S/D process tanks.

9. the pig thrombiase production system described in claim 1, it is characterised in that：Cation exchange area be a CM glucans sun from Son exchanges gel.

10. the pig thrombiase production system described in claim 1, it is characterised in that：Virus filtration area includes connection and is concentrated by ultrafiltration Portion and virus filtration portion, after purifying blood coagulation enzyme solutions are concentrated by ultrafiltration in ultrafiltration concentration portion, virus filtration is carried out through virus filtration portion.